METHODS AND COMPOSITIONS FOR THE TREATMENT OF FOOD FIBERS

专利摘要:
The present invention relates to a method for treating a composition comprising fructan and sucrose, comprising the step of incubating a composition comprising fructan and sucrose, preferably inulin and sucrose, with at least one yeast selected from the group consisting of Saccharomyces, and Kluyveromyces. Incubation with these yeasts leads to the decomposition of the free sugars so that purified fructan compositions are obtained.
公开号:BE1022006B1
申请号:E2014/0598
申请日:2014-08-04
公开日:2016-02-03
发明作者:Robin Denis；Alain Durieux；Christian Fougnies
申请人:Cosucra Groupe Warcoing S.A.；
IPC主号:

专利说明:

METHODS AND COMPOSITIONS FOR THE TREATMENT OF FIBERS
FOOD
FIELD OF THE INVENTION
The present invention relates to methods and compositions for treating dietary fiber. In particular, the present invention relates to methods of purifying fructan-containing compositions by incubating such compositions with yeast to effect degradation and elimination of free sugars.
BACKGROUND OF THE INVENTION
Dietary fiber is edible carbohydrate, which is neither digested nor absorbed in the human small intestine and has been obtained from food material by physical, enzymatic or chemical means and has a beneficial physiological effect. In general, dietary fiber passes through a large part of the digestive system while remaining intact and can be totally or partially fermented by the gut microbiota.
Dietary fiber may be water soluble or insoluble in water. Water-soluble dietary fibers include fructans. Fructans are essentially polymers composed of fructose residues, with or without a glucose unit, which would otherwise be the reducing end. The binding position of the fructose residues determines the type of the fructan. The bond is normally located at one of two primary hydroxyls (OH-1 or OH-6), and there are two basic types of single fructan: inulin (fructosyl residues are linked by ß-2,1 linkages) and levan (the fructosyl residues are linked by β-2,6 linkages). Fructans can be found in many plants as well as microorganisms, where they are stored as energy. For example, inulin is produced in particularly high amounts in chicory roots.
Industrial production of inulin from, for example, chicory root typically involves extraction with hot water. This process produces an extract rich in inulin. However, free sugars (e.g., glucose, fructose and sucrose) are also co-extracted. An inulin-rich extract typically contains about 70 to 85% by weight of inulin and 5 to 13% by weight of free sugars, as well as 10 to 17% by weight of other impurities (eg, salts, proteins, etc.) based on the dry matter. The exact composition, for example, of chicory extracts rich in inulin, however, varies and, for example, depends on the culture conditions, the date of harvest, the variety of chicory, etc.
The process for purifying the inulin-rich extract typically contains several steps, including, for example, solid / liquid separation, ion exchange, activated carbon filtration, etc., in which the majority of the impurities are removed. , and a composition rich in inulin is obtained. However, free sugars which have a structure and / or chemical characteristics very similar to those of fibers such as fructans, in particular inulin, are generally not removed from the inulin-rich composition, and these can represent between 6 and 16% of the dry matter (on the basis of the dry matter, for example 1 to 2% by weight of glucose, 1.5 to 7% by weight of fructose, and 3.5 to 7% by weight of sucrose in inulin-rich compositions from chicory roots).
Although they are physicochemically and structurally similar to fibers, such as fructans, these free sugars are however distinguished by their nutritional properties, given their digestibility which thus confers a high caloric value unlike fibers. High levels of free sugar impurities in dietary fiber compositions is therefore a problem, for example, for diabetics. From this point of view, it is strongly advised to minimize the contents of such free sugars in the inulin-rich composition. In addition, from a technical point of view, industrially produced fibers are often supplied to customers in the form of syrups or powders. In the latter case, the last step of the process can implement spray drying. The effectiveness of this known technique decreases as the content of free sugars increases, these being more "difficult to dry" because of their relative hygroscopicity (mainly fructose), so that the increase in the elimination of sugars Free, and especially fructose, before drying not only has nutritional benefits, but also technical.
There are several ways to separate free sugars from fiber extracts, for example, fractional precipitation based on relative solubility or chromatography. However, these physico-chemical separation techniques are very expensive and have a limited scale performance. The fractional precipitation technique is used industrially for the production of fibers which contain reduced amounts of free sugars. This technique is based on the differential solubility of carbohydrate molecules of different molecular weights. Chromatography on an industrial scale allows the separation of free sugars on different ranges of fiber types and the efficiency is higher than in the case of fractional precipitation, but remains low. In any case, it is not possible to separate the free sugars without suffering a loss of fibers.
In view of the above, there is still a need to develop alternative or improved methods for removing free sugars from dietary fiber compositions, particularly fructan compositions, such as inulin compositions. Therefore, it is an object of the present invention to solve or improve at least one of the disadvantages of the prior art, or to provide a useful alternative.
SUMMARY OF THE INVENTION
The present inventors have unexpectedly discovered that the efficiency, yield, productivity, and / or rate of transformation or purification of fructan can be markedly improved by incubation of a composition comprising fructan and sucrose, preferably, inulin and sucrose, with one or more yeast species, in particular a yeast species selected from the group consisting of or consisting essentially or essentially of Saccharomyces and Kluyveromyces. It has been observed that these yeasts allow a suppression, elimination, reduction or rapid fermentation of free sugars (in particular glucose, fructose and sucrose) of a composition comprising fructan and sucrose, preferably inulin. and sucrose, with a high specificity for free sugars relative to fructan, preferably inulin. Accordingly, the efficiency of the fructan purification, preferably the inulin purification, is increased and the final yield of fructan, preferably inulin, is increased over the treatment of a composition comprising fructan and sucrose without these one or more yeasts. As such, the present process reduces fructan losses, preferably inulin losses, during treatment, such as purification, of such a composition comprising fructan and sucrose, such as chicory extracts. According to the methods as currently described, the treatment (or purification) of a composition comprising fructan and sucrose, preferably inulin and sucrose, leads to a reduction of at least 10% in sugar concentrations free from compositions comprising fructan and sucrose which have not been treated according to the methods of the present invention. In some embodiments, the composition at the end of the process may be, for example, sucrose-free. A particularly advantageous balance between, on the one hand, the specificity of the yeast species described for removing, reducing, eliminating and / or fermenting free sugars, in particular sucrose, but also fructose and glucose, without undesirable fructan degradation , preferably inulin, and on the other hand the rate of removal, reduction, elimination and / or fermentation of free sugars has been observed by the present inventors.
The present invention, in one aspect, therefore relates to a process for transforming, purifying, treating and / or storing a composition comprising fructan and sucrose, preferably inulin and sucrose, comprising incubating a selected yeast. in the group comprising or consisting of or consisting essentially of Saccharomyces and Kluyveromyces with such a composition. In another aspect, the present invention relates to a process for removing, eliminating, reducing, and / or fermenting free sugars (especially glucose, fructose and sucrose) of a composition comprising fructan and sucrose, preferably inulin and sucrose, comprising the step of incubating such a composition with a yeast selected from the group consisting of or consisting of Saccharomyces and Kluyveromyces.
Preferably, the present invention relates to a process for treating a composition comprising fructan and sucrose, comprising the steps of (a) providing a composition comprising fructan and sucrose, said composition comprising fructan and sucrose comprising at least 30% by weight (% w / w) of fructan based on the total dry weight of said composition; and (b) incubating said composition comprising fructan and sucrose with at least one yeast selected from the group consisting of Saccharomyces and Kluyveromyces; until a reduction of at least 10% of the initial weight of sucrose in said composition is obtained.
The present invention further relates to a composition comprising fructan, sucrose and at least one yeast selected from the group consisting of Saccharomyces bayanus, Saccharomyces cerevisiae, Kluyveromyces lactis, and Saccharomyces boulardii, said composition comprising at least 30% by weight (% p / p) fructan based on the total dry weight of said composition.
The present invention further relates to a yeast deposited in the Belgian Coordinated Collections of Microorganisms (BCCM) with the serial number MUCL 55125.
The present invention further relates to the use of a yeast selected from the group consisting of Saccharomyces and Kluyveromyces for reducing the amount of sucrose in a composition comprising sucrose and at least 30% by weight (% w / w) of fructan on the basis of the total weight of dry matter of said composition.
The independent and dependent claims describe particular and preferred features of the invention. The features of the dependent claims may be combined with features of the independent claims or other dependent claims as appropriate. The appended claims are also explicitly included in the description.
The above and other features, components and advantages of the present invention will become apparent upon reading the following detailed descriptions with reference to the accompanying drawings which illustrate, by way of example, the principles of the invention. The reference figures mentioned below designate the appended drawings.
BRIEF DESCRIPTION OF THE FIGURES
Figure 1: represents a graph of growth (measured by optical density at 660 nm) over time at 30 ° C of Saccharomyces cerevisiae w-34/70 incubated with composition B1.
Figure 2: represents a graph of the growth (measured by the optical density at 660 nm) over time at 30 ° C of Kluyveromyces lactis CBS 2103 incubated with the composition B2.
Figure 3: represents a graph of growth (measured by optical density at 660 nm) over time at 30 ° C of Saccharomyces bayanus var. bayanus MUCL 31495 incubated with the composition B3.
Figure 4: represents a graph of growth (measured by optical density at 660 nm) over time at 30 ° C of Saccharomyces bayanus var. uvarum MUCL 31491 incubated with the composition B4.
Figure 5: Graphs of growth (measured by optical density at 660 nm) over time at 30 ° C (A) and at 20 ° C (B) of Saccharomyces bayanus var. uvarum MUCL 55125 incubated with the composition B5.
Figure 6: represents a graph of the evolution of the concentration of free sugars over time at 30 ° C of the composition B1 incubated with Saccharomyces cerevisiae w-34/70. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w based on the total dry matter).
Figure 7: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of the composition B1 incubated with Saccharomyces cerevisiae w-34/70. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) - normalized according to the dilution of the composition).
Figure 8: represents a graph of the evolution of the concentration of free sugars over time at 30 ° C of the composition B2 incubated with Kluyveromyces lactis (CBS 2103). The analyzes are carried out by HPAEC-PAD (results expressed in% w / w based on the total dry matter).
FIG. 9: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C. of the B2 composition incubated with Kluyveromyces lactis (CBS 2103). The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) - normalized according to the dilution of the composition).
Figure 10: represents a graph of the evolution of the concentration of free sugars over time at 30 ° C of the composition B3 incubated with Saccharomyces bayanus var. bayanus MUCL 31495. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w based on the total dry matter).
Figure 11: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of the B3 composition incubated with Saccharomyces bayanus var. bayanus MUCL 31495. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition).
Figure 12: represents a graph of the evolution of the concentration of free sugars over time at 30 ° C of the composition B4 incubated with Saccharomyces bayanus var. uvarum MUCL 31491. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w on the basis of the total dry matter).
Figure 13: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of the composition B4 incubated with Saccharomyces bayanus var. uvarum MUCL 31491. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition).
Figure 14: represents a graph of the evolution of the concentration of free sugars over time at 30 ° C of the composition B5 incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w on the basis of the total dry matter).
Figure 15: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of the B5 composition incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition).
Figure 16: represents a graph of growth (measured by optical density at 660 nm) over time at 30 ° C of Saccharomyces bayanus var. uvarum MUCL 55125 incubated with the composition C.
Figure 17: represents a graph of the evolution of the concentration of free sugars over time at 30 ° C of the composition C incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w).
Figure 18: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of the composition C incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition).
Figure 19: represents a graph of the evolution of the concentration of free sugars over time at 20 ° C of the composition D incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w).
Figure 20: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 20 ° C of the composition D incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition).
Figure 21: represents a graph of the evolution of the concentration of free sugars over time at 20 ° C of the composition E incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w).
Figure 22: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 20 ° C of the composition E incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition).
Figure 23: represents a graph of growth (measured in optical density at 660 nm) over time at 30 ° C of Saccharomyces bayanus var. uvarum MUCL 55125 incubated with the composition A1.
Figure 24: Graphs the evolution of the concentration of free sugars over time at 20 ° C (A) and at 30 ° C (B) of the A1 composition incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w).
Figure 25: Graphs the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 20 ° C (A) and 30 ° C ( B) of the composition A1 incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) - normalized according to the dilution of the composition).
Figure 26: represents a graph of the evolution of the concentration of free sugars over time at 4 ° C of the composition A2 incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w).
Figure 27: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 4 ° C of the A2 composition incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition).
Figure 28: represents a graph of the evolution of the concentration of free sugars over time at 25 ° C with aeration of the composition A3 incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w).
Figure 29: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 25 ° C with aeration of the composition A3 incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) - normalized according to the dilution of the composition).
Figure 30: represents a graph of the evolution of the concentration of free sugars over time at 30 ° C of the composition B6 incubated with Rhodotolula dairenensis (CBS 7294). The analyzes are carried out by HPAEC-PAD (results expressed in% w / w).
Figure 31: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of the composition B6 incubated with Rhodotolula dairenensis (CBS 7294). The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) - normalized according to the dilution of the composition).
Figure 32: represents a graph of the evolution of the concentration of free sugars over time at 30 ° C of the composition B7 incubated with Aureobasidium Pullulans (CBS621.80). The analyzes are carried out by HPAEC-PAD (results expressed in% P / P).
Figure 33: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of the B7 composition incubated with Aureobasidium Pullulans (CBS 621.80). The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) - normalized according to the dilution of the composition).
Figure 34: represents a graph of the evolution of the concentration of free sugars over time at 30 ° C of the composition F incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w).
Figure 35: represents a graph of the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of the F composition incubated with Saccharomyces bayanus var. uvarum MUCL 55125. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition).
DETAILED DESCRIPTION OF THE INVENTION
Before describing the present methods of the invention, it should be understood that this invention is not limited to particular methods, components, products or combinations described, since such methods, components, products and combinations can of course vary. . It should also be understood that the terminology presently used is not intended to be limiting, since the scope of the present invention is limited only by the appended claims.
In the present context, the singular forms "one", "one" and "the" include both singular and plural forms unless otherwise indicated in the context.
The terms "comprising", "includes" and "consisting of" in this context are synonymous with "comprising", "includes" or "containing", "contains", and are inclusive or open and do not exclude additional members, elements or process steps not mentioned. It will be apparent that the terms "comprising", "includes" and "consisting of", in this context, include the terms "consisting of", "consists of" and "consists of", and the terms "consisting primarily of "Essentially consists" and "essentially consists of".
The reference to digital ranges by terminals includes all the numbers and fractions included in the respective ranges, as well as the mentioned terminals.
The term "about" or "approximately", in this context, with reference to a measurable value such as a parameter, a quantity, a time duration, and the like, is intended to include variations of +/- 20% or less, preferably +/- 10% or less, more preferably +/- 5% or less, and still more preferably +/- 1% or less of and in relation to the specified value, to the extent that such variations are suitable for operating in the described invention. It should be understood that the value at which the "about" or "approximately" modifier is itself also specifically, and preferably, described.
Whereas the terms "one or more" or "at least one", such as one or more or at least one member (s) of a group of members, are clear as such, as an additional example the term includes inter alia a reference to any one of said members, or to any two or more of said members, such as, for example, any of £ 3, £ 4,> 5,> 6 or> 7 etc., said members, and up to all of said members.
All references cited in this specification are presently incorporated by reference in their entirety. In particular, the teachings of all the references currently specifically referenced are incorporated by reference.
Unless defined otherwise, all terms used in the description of the invention, including technical and scientific terms, have the definition as commonly understood by those skilled in the art to which this invention belongs. As another recommendation, definitions of terms are included to better appreciate the teaching of the present invention.
In the following passages, various aspects of the invention are defined in more detail. Each aspect defined in this way can be combined with another aspect or any other aspect, unless otherwise clearly indicated in the context. In particular, a feature indicated as being preferred or advantageous may be combined with another feature or other features indicated as preferred or advantageous.
Any reference in this specification to "an embodiment" means that a particular / particular element, structure or feature described with reference to the embodiment is included in at least one embodiment of the present invention. Therefore, aspects of the phrase "in one embodiment" at different locations in this specification do not necessarily refer to the same embodiment, although this is possible. In addition, the particular elements, structures or features may be combined in any suitable manner, as will be apparent to those skilled in the art of this description, in one or more embodiments. In addition, although some embodiments described herein include some, but not other elements included in other embodiments, combinations of elements of different embodiments are intended to be within the scope of the invention. , and form different embodiments, as will be apparent to those skilled in the art. For example, in the appended claims, any of the claimed embodiments may be used in any combination.
In the following detailed description of the invention, reference is made to the accompanying drawings which form a part thereof, and in which are illustrated by way of illustration only specific embodiments in which the invention can be implemented. convenient. It should be understood that other embodiments may be used and structural or logical changes may be made without departing from the scope of the present invention. The detailed description given below therefore should not be considered in a limiting sense, and the scope of the present invention is defined by the appended claims.
Preferred aspects (features) and embodiments of this invention are presently defined below. Each aspect and embodiment of the presently defined invention may be combined with another aspect and / or other embodiments unless otherwise clearly indicated in the context. In particular, any feature indicated as being preferred or advantageous may be combined with another feature or other features or aspects indicated as being preferred or advantageous. Hereinafter, the present invention is presented in particular by any one or any combination of one or more of the aspects and embodiments numbered below from 1 to 66, with another aspect and / or other embodiments. A process for treating a composition comprising fructan and sucrose, comprising the steps of (a) providing a composition comprising fructan and sucrose, wherein said composition comprising fructan and sucrose comprises at least 30% by weight (% w / w) fructan based on the total dry weight of said composition; and (b) incubating said composition comprising fructan and sucrose with at least one yeast selected from the group consisting of Saccharomyces and Kluyveromyces; until a reduction of at least 10% of the initial weight of sucrose in said composition is obtained. 2. The method according to aspect 1, wherein said at least one yeast is selected from the group consisting of Saccharomyces bayanus, Kluyveromyces lactis, Saccharomyces cerevisiae and Saccharomyces boulardii, preferably selected from the group consisting of Saccharomyces bayanus, and Kluyveromyces lactis. A process according to any one of claims 1 or 2, wherein said composition comprising fructan and sucrose, at the beginning of the incubation, further comprises one or more free sugars (other than sucrose), preferably in wherein said free sugars are selected from the group consisting of glucose and fructose. A process according to any one of claims 1 to 3 wherein said composition comprising fructan and sucrose at the beginning of the incubation further comprises one or more free sugars selected from the group consisting of, consisting of or substantially consisting of glucose and fructose. A process according to any one of claims 1 to 4, wherein said composition comprising fructan and sucrose at the beginning of the incubation comprises at least 1% by weight of free sugars on the basis of the total weight of material dry of the composition. A process according to any one of claims 1 to 5, wherein said composition comprising fructan and sucrose, at the beginning of the incubation, comprises at most 70% by weight of free sugars on the basis of the total weight of material dry of the composition. 7. Process according to any one of the aspects 1 to 6, in which said fructan has a number of average polymerization degree (DP) of at least 3, for example at least 5, for example at least 7, for example at at least 10, for example at least 15, for example at least 20, for example at least 25, for example at least 70. 8. Process according to any one of aspects 1 to 7, in which said fructan has a mean DP in number in the range of 3 to 30. 9. A process according to any one of claims 1 to 8, wherein said fructan is of plant origin, preferably from chicory. The process of any one of claims 1 to 9, wherein said fructan is a chicory fructan. 11. Process according to any one of the aspects 1 to 10, wherein said fructan is inulin, preferably chicory inulin. The method of any one of claims 1 to 11, wherein said fructan is an inulin having a DP in the range of 2 to about 100. A process according to any one of claims 1 to 12, wherein said fructan is an inulin having a formula GFn and / or Fm, wherein G is a glucose unit, F is a fructose unit, n is an integer representing the number of fructose units linked to the terminal glucose unit, and m is an integer representing the number of mutually linked fructose units in the carbohydrate chain, wherein n is at least 2, and m is at least 2. 14. The process of any one of aspects 1 to 13, wherein said fructan is partially hydrolysed. . A process according to any one of claims 1 to 14, wherein said fructan comprises or consists of or consists essentially of fructo-oligosaccharides. A process according to any one of claims 1 to 15, wherein said fructan comprises or consists of or consists essentially of fructo-oligosaccharides, and said fructo-oligosaccharides have a number average DP of at least 3 and at most 7 17. The process according to any of claims 1 to 16, wherein said composition comprising fructan and sucrose comprises at least 40% by weight of fructan based on the total dry weight of the composition, preferably less than 50% by weight, still more preferably at least 60% by weight. 18. A process according to any of claims 1 to 17, wherein said composition comprising fructan and sucrose comprises at most 99% by weight of fructan based on the total dry weight of the composition. 19. A process according to any one of claims 1 to 18, wherein said composition comprising fructan and sucrose comprises at least 30% by weight and at most 99% by weight of fructan based on the total dry matter weight of the composition, preferably at least 40% by weight, preferably at least 50% by weight, still more preferably at least 60% by weight. 20. Process according to any one of the aspects 1 to 19, wherein said composition comprising fructan and sucrose is a liquid composition, preferably an aqueous composition. A process according to any one of claims 1 to 20, wherein said composition comprising fructan and sucrose comprises at least 5% by weight of dry matter based on the total weight of the composition, preferably at least 8% by weight, preferably at least 10% by weight. 22. Process according to any one of the aspects 1 to 21, wherein said composition comprising fructan and sucrose comprises at least 5% by weight and at most 80% by weight of dry matter on the basis of the total weight of the composition for example at least 8% by weight, preferably at least 10% by weight, preferably at most 70% by weight, preferably at most 60% by weight, preferably at most 55% by weight, preferably at most 50% by weight. 23. Process according to any one of the aspects 1 to 22, in which the said composition comprising fructan and sucrose comprises at most 80% by weight of dry matter on the basis of the total weight of the composition, preferably at most 70%. by weight, preferably at most 60% by weight, preferably at most 55% by weight, preferably at most 50% by weight. The method of any one of claims 1 to 23, further comprising the step of adding a nitrogen source to said composition comprising fructan and sucrose, before and / or during step (b). ), preferably the addition of yeast extract. 25. A process according to any of claims 1 to 24, further comprising one or both of aeration and stirring steps of the composition comprising fructan and sucrose during incubation with said yeast. Process according to any one of aspects 1 to 25, wherein said composition comprising fructan and sucrose is incubated with said yeast at a temperature of at least the freezing point of the composition, preferably above freezing point of said composition. 27. A process according to any one of aspects 1 to 26, wherein said composition comprising fructan and sucrose is incubated with said yeast at a temperature of at least -5 ° C, preferably at least 0 ° C, by at least 5 ° C, for example at least 10 ° C, for example at least 15 ° C, for example at least 20 ° C. 28. Process according to any one of the aspects 1 to 27, wherein said composition comprising fructan and sucrose is incubated with said yeast at a temperature of at most 40 ° C, for example at most 35 ° C, for example at most 30 ° C. 29. Process according to any one of aspects 1 to 28, wherein said composition comprising fructan and sucrose is incubated with said yeast at a temperature of at least the freezing point of said composition and at most 40 ° C, for example at least -5 ° C and at most 40 ° C, for example at least 0 ° C and at most 35 ° C, for example at least 5 ° C and at most 33 ° C, for example at least 10 And at most 30 ° C, for example at least 15 ° C and at most 30 ° C, for example at least 20 ° C and at most 30 ° C. 30. The method of any one of aspects 1 to 29, wherein said composition comprising fructan and sucrose is incubated with said yeast until a reduction of at least 20% of the initial weight of free sugars in said composition is obtained, preferably a reduction of at least 30%, for example at least 40%, for example at least 50%; preferably at least 60%, for example at least 70%, for example at least 80%, preferably at least 90%, preferably at least 95%, for example at least 98%, for example a reduction of at least 99%. 31. A process according to any of claims 1 to 30, wherein said composition comprising fructan and sucrose is incubated with said yeast for at least 5 hours, preferably for at least 10 hours. 32. The method of any one of claims 1 to 31, wherein said composition comprising fructan and sucrose is incubated with said yeast for not more than 12 months. 33. Process according to any one of aspects 1 to 32, wherein said composition comprising fructan and sucrose is incubated with said yeast at a pH of at least 2.5, preferably a pH of at least 3, 0, preferably a pH of at least 3.5. 34. Process according to any one of aspects 1 to 33, wherein said composition comprising fructan and sucrose is incubated with said yeast at a pH of at most 8.5, for example at a pH of at most 8. , 0, for example at a pH of at most 7.5, for example at a pH of at most 7.0. 35. A method according to any one of the aspects 1 to 34, comprising incubating at the beginning of said incubation at least 103 colony-forming units (CFU) per ml, for example at least 104 CFU of said yeast per ml of said composition comprising fructan and sucrose. 36. A method according to any one of aspects 1 to 35, comprising incubating at the beginning of said incubation of at most 1010 CFU / ml, for example at most 109 CFU / ml, for example at most 108 CFU of said yeast. per ml of said composition comprising fructan and sucrose. 37. The method according to any one of the aspects 1 to 36, wherein said composition comprising fructan and sucrose, at the beginning of the incubation, comprises at least 1% by weight and at most 70% by weight of free sugars comprising said sucrose on the basis of the total dry weight of the composition. 38. Process according to any one of the aspects 1 to 37, wherein the composition of step (a) is obtained by extraction with hot water of a material containing fructans. 39. The process according to aspect 38, wherein said fructan-containing material is of plant origin. 40. The method according to aspect 38 or 39, wherein said fructan-containing material is chicory. 41. Process according to any one of the aspects 1 to 40, wherein the composition of step (a) is obtained by means of a process comprising the steps of (i) extraction with hot water of a fructan-containing material, and (ii) filtering the hot water extract so as to recover a composition comprising fructan and sucrose from step (a). 42. Process according to any one of the aspects 1 to 41, wherein the composition of step (a) is obtained by means of a process comprising the steps of (i) extraction with hot water of a material containing fructans, (ii) filtration of the hot water extract; and (iii) demineralizing the filtrate of step (ii) so as to recover a composition comprising fructan and sucrose from step (a). 43. Process according to any one of the aspects 1 to 42, wherein the composition of step (a) is obtained by means of a process comprising the steps of (i) extraction with hot water of a material containing fructans, (ii) filtration of the hot water extract; (iii) demineralization of the filtrate of step (ii); and (iv) active carbon filtration of the filtrate of step (iii) so as to recover a composition comprising fructan and sucrose from step (a). 44. The process of any one of claims 1 to 43, further comprising the step of removing said yeast after incubation with said composition comprising fructan and sucrose. 45. Process according to any one of the aspects 1 to 44, wherein said yeast is a lysate of said yeast or an extract of said yeast. 46. A process according to any one of claims 1 to 45, wherein the weight ratio of the free sugars comprising fructan sucrose in said composition comprising fructan and sucrose at the beginning of the incubation is at least 1: 100. A process according to any one of claims 1 to 46, wherein the weight ratio of free sugars comprising fructan sucrose in said composition comprising fructan and sucrose at the beginning of the incubation is at most 2, 3: 1. 48. A process according to any of claims 1 to 47, wherein the weight ratio of the free sugars comprising fructan sucrose in said composition comprising fructan and sucrose at the beginning of the incubation is at least 1: 100 and at most 2.3: 1 49. A method according to any one of claims 1 to 48, wherein Saccharomyces is Saccharomyces bayanus var. uvarum deposited in the Belgian Coordinated Collections of Microorganisms (BCCM) with the serial number MUCL 55125. 50. The method of any one of aspects 1 to 49, wherein at the end of said incubation step, the The fructan weight of said composition comprising fructan and sucrose is less than 20% by weight of the initial fructan weight at the beginning of said incubation, preferably at most 10%, most preferably at most 5%. 51. A process according to any one of claims 1 to 50, wherein said composition comprising fructan and sucrose is incubated with said yeast until a reduction of at least 20% of the initial weight of free sugars (comprising sucrose) in said composition is obtained, preferably a reduction of at least 30%, for example at least 40%, for example at least 50%, for example at least 60%; preferably at least 70%, for example at least 80%, for example at least 90%, preferably at least 95%, for example at least 98%, for example at least 99%, and the fructan weight of said composition comprising fructan and sucrose is less than 20% of the initial weight of fructan weight at the beginning of said incubation, preferably at most 10%, most preferably at most 5%. 52. A process according to any one of claims 1 to 51, wherein said composition comprising fructan and sucrose is incubated with said yeast until a reduction of at least 50% of the initial weight of free sugars ( comprising sucrose) in said composition is preferably obtained a reduction of at least 60%, for example at least 70%, for example at least 80%, preferably at least 90%, preferably at least 95%, for example at least 98%, for example at least 99%, and the fructan weight of said composition comprising fructan is at most 5% lower than the initial weight of fructan at the beginning of said incubation. 53. The method of any one of aspects 1 to 52, wherein said at least one yeast is selected from the group consisting of or consisting of Saccharomyces and Kluyveromyces; preferably, said at least one yeast is selected from the group consisting of or consisting of Saccharomyces bayanus, Saccharomyces cerevisiae, Kluyveromyces lactis, and Saccharomyces boulardii; still more preferably said at least one yeast is selected from the group consisting of or consisting of Saccharomyces bayanus var. uvarum, Saccharomyces bayanus var. bayanus, Saccharomyces cerevisiae, Saccharomyces boulardii, and Kluyveromyces lactis var. drosophylarum, still more preferably said at least one yeast is selected from the group consisting of or consisting of
Saccharomyces bayanus var. uvarum MUCL 55125 (deposited at BCCM / MUCL Louvain-La-Neuve); Saccharomyces bayanus MUCL 31491 (obtained from BCCM / MUCL Louvain-La-Neuve), Saccharomyces bayanus MUCL 31495 (obtained from BCCM / MUCL Louvain-La-Neuve), Saccharomyces bayanus BC S103 (obtained from Fermentis, Lesaffre group), Saccharomyces bayanus VR 44 (obtained from Fermentis, Lesaffre group), Kluyveromyces lactis var. drosophylarum CBS 2103 (obtained from the CBS-KNAW Fungal Biodiversity Center, Utrecht, the Netherlands), Saccharomyces cerevisiae w-34/70 (obtained from Fermentis, Lesaffre group), and Saccharomyces boulardii (obtained from Enterol®; biocodex gamma). 54. The method of any one of aspects 1 to 53, wherein said at least one yeast is selected from the group consisting of or consisting of
Saccharomyces; preferably, said at least one yeast is selected from the group consisting of or consisting of Saccharomyces bayanus, Saccharomyces cerevisiae and Saccharomyces boulardii; still more preferably, said at least one yeast is selected from the group consisting of or consisting of Saccharomyces bayanus var. uvarum, Saccharomyces bayanus var. bayanus, Saccharomyces cerevisiae, Saccharomyces boulardii and even more preferably said at least one yeast is selected from the group consisting of or consisting of Saccharomyces bayanus var. uvarum MUCL 55125 (deposited at BCCM / MUCL Louvain-La-Neuve); Saccharomyces bayanus MUCL 31491 (obtained from BCCM / MUCL Louvain-La-Neuve), Saccharomyces bayanus MUCL 31495 (obtained from BCCM / MUCL Louvain-La-Neuve),
Saccharomyces bayanus BC S103 (obtained from Fermentis, Lesaffre group), Saccharomyces bayanus VR 44 (obtained from Fermentis, Lesaffre group), Saccharomyces cerevisiae w-34/70 (obtained from Fermentis, group
Lesaffre), and Saccharomyces boulardii (obtained from Enterol®, biocodex gamma), and even more preferably Saccharomyces bayanus var. uvarum MUCL 55125 (deposited at the BCCM / MUCL Louvain-La-Neuve). 55. A composition comprising fructan, sucrose and at least one yeast selected from the group consisting of Saccharomyces bayanus, Saccharomyces cerevisiae, and Kluyveromyces lactis. 56. A composition comprising fructan, sucrose and at least one yeast selected from the group consisting of Saccharomyces bayanus, Saccharomyces cerevisiae, and Kluyveromyces lactis, said composition comprising sucrose and at least 30% by weight of fructan based on total weight dry matter of said composition. 57. Yeast deposited in the Belgian Coordinated Collections of Microorganisms (BCCM) with accession number MUCL 55125. 58. Use of yeast in aspect 57 to reduce the amount of free sugars comprising sucrose in a composition comprising fructan and sucrose, more preferably for reducing the amount of sucrose in a composition comprising sucrose and at least 30% by weight of fructan based on the total dry weight of said composition. 59. Use of a yeast selected from the group consisting of Saccharomyces and Kluyveromyces for reducing the amount of free sugars comprising sucrose in a composition comprising fructan and sucrose, more preferably for reducing the amount of sucrose in a composition comprising sucrose and at least 30% by weight of fructan based on the total dry weight of said composition. A method for reducing the amount of free sugars comprising sucrose in a composition comprising fructan and sucrose comprising the step of using the process according to any one of aspects 1 to 54. 61. Process for purifying a composition comprising fructan and sucrose comprising the step of using the process according to any one of the aspects 1 to 54. 62. A method for storing a composition comprising fructan and sucrose comprising the step of using the method according to any of aspects 1 to 54. 63. A process for treating a composition comprising fructan and sucrose comprising the step of using the process according to any one of aspects 1 to 54. 64. Process for removing free sugars, from a composition comprising fructan and sucrose comprising the step of using the method according to any one of the aspects 1 to 54. 65. Procé for reducing the amount of free sugars from a composition comprising fructan and sucrose comprising the step of using the process according to any one of the aspects 1 to 54. 66. Process for removing free sugars, from a composition comprising fructan and sucrose comprising the step of using the method according to any one of the aspects 1 to 54. 67. Process for fermenting free sugars from a composition comprising fructan and sucrose comprising the step of using the method according to any one of aspects 1 to 54. 68. Use of yeast selected from the group consisting of Saccharomyces, and Kluyveromyces for reducing the amount of free sugars comprising sucrose in a composition comprising fructan and sucrose, more preferably for reducing the amount of sucrose in a composition comprising sucrose and at least 30% by weight of fructan based on the total weight of dry matter of said composition or any one or more of the aspects 1 to 54.
In a first aspect, the present invention relates to a process for treating a composition comprising fructan and sucrose, preferably inulin and sucrose, comprising the step of incubating a composition comprising fructan and sucrose, said composition comprising fructan and sucrose comprising at least 30% by weight of fructan based on the total dry weight of said composition, with at least one yeast selected from the group consisting of or consisting of Saccharomyces and Kluyveromyces, up to a reduction of at least 10% of the initial weight of sucrose in said composition is obtained; as well as the combined or separate use of these one or more yeasts for the application mentioned above. Said reduction of at least 10% of the initial weight of sucrose in said composition can be measured by liquid chromatography such as, for example, by high performance anion exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD).
In another aspect, the present invention relates to a process for purifying a composition comprising fructan and sucrose, preferably inulin and sucrose, comprising the step of incubating a composition comprising fructan and sucrose, said composition comprising fructan and sucrose comprising at least 30% by weight of fructan based on the total dry weight of said composition, with at least one yeast selected from the group consisting of or consisting of Saccharomyces and Kluyveromyces until a reduction of at least 10% of the initial weight of sucrose in said composition is obtained; as well as the combined or separate use of these one or more yeasts for the application mentioned above.
In another aspect, the present invention relates to a process for treating a composition comprising fructan and sucrose, preferably inulin and sucrose, comprising the step of incubating a composition comprising fructan and sucrose, said composition comprising fructan and sucrose comprising at least 30% by weight of fructan based on the total dry weight of said composition, with at least one yeast selected from the group consisting of or consisting of Saccharomyces and Kluyveromyces until a reduction of at least 10% of the initial weight of sucrose in said composition is obtained; as well as the combined or separate use of these one or more yeasts for the application mentioned above.
In another aspect, the present invention relates to a method for storing a composition comprising fructan and sucrose, preferably inulin and sucrose, comprising the step of incubating a composition comprising fructan and sucrose, said composition comprising fructan and sucrose comprising at least 30% by weight of fructan based on the total dry weight of said composition, with at least one yeast selected from the group consisting of or consisting of Saccharomyces and Kluyveromyces until a reduction of at least 10% of the initial weight of sucrose in said composition is obtained; as well as the combined or separate use of these one or more yeasts for the application mentioned above, and the storage of said composition.
In another aspect, the present invention relates to a method for removing carbohydrates, preferably free sugars comprising sucrose, more preferably carbohydrate monomers and / or carbohydrate dimers, most preferably monomers or dimers of hexose and / or pentose, most preferably sucrose, glucose and / or fructose, a composition comprising fructan and sucrose, preferably inulin and sucrose, comprising the incubation step of a composition comprising fructan and sucrose, said composition comprising fructan and sucrose comprising at least 30% by weight of fructan based on the total dry weight of said composition, with at least one yeast selected from the group comprising or consisting of Saccharomyces and Kluyveromyces until a reduction of at least 10% of the initial weight of sucrose in said composition n is obtained; as well as the combined or separate use of these one or more yeasts for the application mentioned above.
In another aspect, the present invention relates to a method for reducing the amount of sugars, preferably free sugars, more preferably carbohydrate monomers and / or carbohydrate dimers, most preferably hexose monomers or dimers. and / or pentose, most preferably sucrose, glucose and / or fructose, in a composition comprising fructan and sucrose, preferably inulin and sucrose, comprising the incubation step of a composition comprising fructan and sucrose, said composition comprising fructan and sucrose comprising at least 30% by weight of fructan based on the total dry weight of said composition, with at least one yeast selected from the group consisting of or consisting of Saccharomyces and Kluyveromyces until a reduction of at least 10% of the initial weight of sucrose in said composition is obtained ; as well as the combined or separate use of these one or more yeasts for the application mentioned above.
In another aspect, the present invention relates to a process for removing sugars, preferably free sugars, more preferably carbohydrate monomers and / or carbohydrate dimers, most preferably hexose monomers or dimers and / or pentose, most preferably sucrose, glucose and / or fructose, a composition comprising fructan and sucrose, preferably inulin and sucrose, comprising the step of incubating a composition comprising fructan and sucrose, said composition comprising fructan and sucrose comprising at least 30% by weight of fructan based on the total dry weight of said composition, with at least one yeast selected from the group consisting of or consisting of Saccharomyces and Kluyveromyces until a reduction of at least 10% of the initial weight of sucrose in said composition is obtained; as well as the combined or separate use of these one or more yeasts for the application mentioned above.
In another aspect, the present invention relates to a process for fermenting sugars, preferably free sugars, more preferably carbohydrate monomers and / or carbohydrate dimers, most preferably hexose and / or pentose monomers or dimers. , most preferably sucrose, glucose and / or fructose, in a composition comprising fructan and sucrose, preferably inulin and sucrose, comprising the step of incubating a composition comprising fructan and sucrose, said composition comprising fructan and sucrose comprising at least 30% by weight of fructan based on the total dry weight of said composition, with at least one yeast selected from the group consisting of or consisting of Saccharomyces and Kluyveromyces until a reduction of at least 10% of the initial weight of sucrose in said composition is obtained; as well as the combined or separate use of these one or more yeasts for the application mentioned above.
In the present context, the term "incubation" or "incubated" refers to the contacting of the yeast as presently described with the composition comprising fructan and sucrose, preferably inulin and sucrose, as presently described. and preferably maintaining such a mixture under specific conditions to promote a particular reaction, especially fermentation. It should be understood that if live yeasts are used, the incubation parameters are defined so that viability of the yeast, at least for a specified time, is ensured.
In the present context, the term "fructan" refers to polymers of fructose molecules. A glucose pattern at what would otherwise be the reducing end may be present in fructans. The position of the linker of the fructose residues can determine the type of the fructan. The bond may be present at one of the two primary hydroxyls (OH-1 or OH-6). Fructan for use for use in the present invention comprises both basic types of simple fructan: inulins (in which fructosyl residues are generally bound by β-2,1 linkages) and levans (in which fructosyl residues are generally linked by β-2,6 linkages). It is presently included graminiane or mixed fructans which have both β-2.1 and β-2,6 linkages between the fructose units, and therefore contain branching. In plants, up to 1000 fructose units can be linked in a single fructan molecule. The fructan for use in the present invention may also include microbial fructans which may include up to 100,000 fructose units. Fructan for use in the present invention can be found in plants, algae and bacteria. Fructans are a type of dietary fiber. The fructans for use in the present invention can be mainly obtained industrially from chicory roots (Cichorium intybus) or from Jerusalem artichoke (Helianthus tuberosus). Inulin degradation products are fructo-oligosaccharides (FOS), i.e. hydrolysis of inulin can produce fructooligosaccharides, which are oligomers having a DP generally below 20. , which are also understood now. Fructo-oligosaccharides can also be synthesized enzymatically from sucrose.
As used herein, the term "inulin" refers to a mixture of oligo- and / or polysaccharides of fructose that may have terminal glucose. Inulins belong to a class of fibers called fructans. In one embodiment, the inulin may be represented, according to the terminal carbohydrate moiety, by the general formulas GFn and / or Fm, where G is a glucose unit, F is a fructose unit, n is an integer representing number of fructose units linked to the terminal glucose unit, and m is an integer representing the number of mutually linked fructose units in the carbohydrate chain, preferably where n is at least 2, and m is at least 2. Inulin for use in the present invention include inulins with a terminal glucose which are also referred to as alpha-D-glucopyranosyl- [beta-D-fructofuranosyl] (n-1) -D-fructofuranosides, as well as glucose-free inulins which are also referred to as beta -D-fructopyranosyl- [D-fructofuranosyl] (n-1) -D-fructofuranosides. Inulins for use in the present invention may also include a branched inulin. Inulins for use in the present invention may also include inulin hydrolysis products such as fructo-oligosaccharides (FOS), also referred to as oligofructoses, which are oligomers of fructose with a DP of 20, and may also include fructo-oligosaccharides ending in terminal glucose with a DP of 3 to 5 synthesized from sucrose. Preferably, said fructo-oligosaccharides have a number average DP of at least 3 and at most 7. Vegetable-based inulin-adapted saccharide chains for use in the invention can have a DP in the range of 2. at about 100. The inulin can be a liquid or powder product.
In the present context, the terms "degree of polymerization" or "(DP)" refer to the number of monosaccharide residues present in an oligo- or polysaccharide. In addition, the average degree of polymerization parameter is often used. The degree of polymerization is a measurement of molecular weight (MW). DP can be calculated as the ratio of the total MW of the polymer or oligomer to the MW of the repeating units.
The average degree of polymerization (av DP) of a mixture of oligo- or polysaccharides (polydispersed) is the average of the degree of polymerization (DP) of all the molecules present in this saccharide mixture. In the present context, the average degree of polymerization, unless otherwise indicated, is calculated on the basis of the number of molecules for each DP: av DPn or number-average degree of polymerization as hereinafter described.
Determination of the molecular weight distribution of the fructan sample is performed by high performance anion exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD) on a Thermo Scientific - Dionex ICS 5000 chromatography system. Separation of the different chain lengths is carried out with a Carbopac PA100 4 mm x 250 mm column (+ precolumn) at 40 ° C. with a flow rate of 1 ml / min. 160 mM sodium hydroxide is used as the eluent. A sodium acetate gradient during the test makes it possible to separate the different lengths of the chain.
Standards of fructan mixtures at different concentrations are injected to plot the calibration curves and assign the peaks in the chromatogram based on the retention time of the standard. The calibration curves make it possible to determine the concentration of each molecular species in the sample. From the concentration distribution obtained, the number-average degree of polymerization Dpn is calculated by - ZjNtDpt where Ni is the number of molecules with residues and Dpi the number of residues.
In one embodiment, the fructan as described herein, preferably inulin, has a number average DP of at least 3. In one embodiment, the fructan as described herein, preferably inulin, has a number average DP of at most 500. In one embodiment, said fructan, preferably inulin, has a number average DP of at least 3, for example at least 5, for example at least 7. , for example at least 10, for example at least 15, for example at least 20, for example at least 25, for example at least 70. In one embodiment, the fructan as described above, preferably inulin, has a mean DP in number of at least 3 and at most 500, preferably at least 3 and at most 100, more preferably at least 3 and at most 30. In another embodiment preferred embodiment, the fructan as currently described, preferably inulin, comprises or consists of fructo-oli gosaccharides (FOS). In another preferred embodiment, the fructan as described herein has a number average DP of at least 3 and at most 20, preferably at least 3 and at most 15, e.g. at least 3 and at most 10. In yet another preferred embodiment, the fructan as described herein, preferably inulin, comprises or consists of fructan, preferably hydrolyzed or partially hydrolysed inulin. Hydrolyzed fructan, such as hydrolyzed inulin, may, for example, be obtained enzymatically (for example, by inulinases) or alternatively may be obtained by acid and / or thermal hydrolysis.
In one embodiment, fructan, preferably inulin, as described herein is derived or isolated from plants, i.e., of plant origin, preferably chicory (Cichorium intybus). ), agave (Agave spp.), banana (Musa spp.), burdock (Arctium lappa), camassia (Camassia spp.), Echinacea (Echinacea spp.), Saussurea Costus lappa, dandelion (Taraxacum ruderalia), large elk (Inula helenium), garlic (Allium sativum), Jerusalem artichoke (Helianthus tuberosus), jicama (Pachyrhizus erosus), arnica (Arnica montana), mugwort (Artemisia vulgaris), onion (Allium cepa), wild yam (Dioscorea spp.), earth pear (Smallanthus sonchifolius spp.), Leek (Allium porum), asparagus, Scorzonera hispanica, salsify (Tragopogon porrifolius), wheat (Tritichum aestivum), dahlia (Dahlia spp.), Most preferably chicory.
In one embodiment, the composition comprising fructan and sucrose is obtained by extraction with hot water. In a preferred embodiment, the industrial production of fructan, such as inulin of, for example, chicory root implements extraction with hot water. However, free sugars (such as glucose, fructose and sucrose) are co-extracted.
In the present context, the term "free sugars" refers to monosaccharides and / or disaccharides. Free sugars can, for example, be present in plants, plant material, or homogenates, plant extracts or isolates, or fractionated plant material. In a preferred embodiment, the term "free sugars" as used herein refers to mono- or di-saccharides of hexose or pentose, preferably hexose mono- or di-saccharides. Most preferably, the term "free sugars" includes fructose, glucose, and sucrose (sucrose). Accordingly, in one embodiment, the free sugars comprise or consist of or consist essentially of fructose. In another embodiment, the free sugars comprise or consist of or consist essentially of glucose. In another further embodiment, the free sugars comprise or consist of or consist essentially of sucrose. In another embodiment, the free sugars comprise or consist of fructose and glucose. In another further embodiment, the free sugars comprise or consist of or consist essentially of fructose and sucrose. In another embodiment, the free sugars comprise or consist of or consist essentially of glucose and sucrose. In another further embodiment, the free sugars comprise or consist of or consist essentially of fructose, glucose, and sucrose.
In embodiments, in the methods as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, further comprises one or more free sugars, defined as described above.
As used herein, the term "composition comprising fructan and sucrose" or "composition comprising inulin and sucrose" refers to any type of composition that contains fructan or inulin respectively and sucrose. Such a composition may be a dry composition. Preferably, such a composition is a liquid composition, most preferably an aqueous composition (i.e., a composition comprising water and a certain amount of fructan, preferably inulin, dissolved and or dispersed therein). The compositions can be obtained by homogenization, for example, of plant material. Preferably, the compositions as presently described refer to extracts, which are enriched in fructan, preferably inulin, with respect to the source material from which they are derived. Inulin extraction may, for example, implement placement of plant material in hot water followed by concentration (e.g., evaporation). In one embodiment, the compositions comprising fructan and sucrose, preferably inulin and sucrose, as presently described comprise at least 30% by weight of fructan, preferably inulin, based on the weight total dry matter content of the composition, preferably at least 40% by weight, preferably at least 50% by weight, preferably at least 60% by weight of fructan, for example at least 30 g of fructan, preferably inulin, per 100 g of dry matter. In one embodiment, the compositions comprising fructan and sucrose, preferably inulin and sucrose, as presently described comprise at least 1.5% by weight of fructan based on the total weight of the composition, preferably inulin; preferably at least 5.0% by weight of fructan, preferably inulin; more preferably at least 8.0% by weight of fructan, preferably inulin. In another embodiment, the compositions comprising fructan and sucrose, preferably inulin and sucrose, as presently described comprise at most 80% by weight of fructan; preferably inulin, based on the total weight of the composition. In one embodiment, the compositions comprising fructan and sucrose, preferably inulin and sucrose, as presently described comprise at most 70% by weight, for example at most 60% by weight, for example at most 50% by weight of fructan, preferably inulin; for example at most 45% by weight of fructan, preferably inulin based on the total weight of the composition. In a preferred embodiment, the compositions comprising fructan and sucrose, preferably inulin and sucrose, as presently described comprise at least 1.5% by weight of fructan and at most 80% by weight of fructan. , preferably inulin, that is to say at least 1.5 g and at most 80 g of fructan, preferably inulin per 100 g of composition. In one embodiment, the compositions comprising fructan and sucrose, preferably inulin and sucrose, as presently described comprise at least 5% by weight and at most 70% by weight of fructan based on weight. total composition; preferably inulin, preferably at least 8% by weight and at most 65% by weight of fructan, preferably inulin; more preferably at least 8% by weight and at most 50% by weight of fructan, preferably inulin; still more preferably at least 8% by weight and at most 45% by weight of fructan, preferably inulin.
The composition comprising fructan and sucrose, such as an extract rich in inulin, can be obtained by extraction with hot water of a plant material. The plant material, for example, chicory roots is first harvested and can then be washed and, if necessary, cut into chips (strips or slices). Extraction with hot water can be carried out by countercurrent diffusion with hot water of the plant material, preferably sliced plant material. A typical ratio of vegetable material (for example chips) to water may be, for example, 1. A suitable temperature may be at least 50 ° C, for example at least 60 ° C, for example at least 70 ° C. A typical extraction time can vary from 1 to 10 hours. The resulting juice containing fructan in solution can, if necessary, be coarsely filtered to remove the spent plant material.
Preferably, the composition of step (a) is obtained using a process comprising the steps of (i) extracting with hot water a material containing fructans, (ii) filtering the extract of Hot water ; and (iii) demineralizing the filtrate of step (ii) so as to recover a composition comprising fructan and sucrose from step (a). In one embodiment, the composition of step (a) is obtained using a method comprising the steps of (i) extracting with hot water a material containing fructans, (ii) filtering the hot water extract; (iii) demineralization of the filtrate of step (ii); and (iv) active carbon filtration of the filtrate of step (iii) so as to recover a composition comprising fructan and sucrose from step (a).
An example of a resulting composition comprising fructan and sucrose, preferably inulin and sucrose, can have a typical solids content of 13% and comprise about 77% by weight of inulin on a dry matter basis. and about 9% by weight of free sugars (including sucrose).
The yeasts that can be used in the methods as described herein are selected from the group consisting of or consisting of Saccharomyces and Kluyveromyces; preferably, the yeasts are selected from the group consisting of or consisting of Saccharomyces bayanus; Saccharomyces cerevisiae; Kluyveromyces lactis, and Saccharomyces boulardii; still more preferably the yeasts are selected from the group consisting of or consisting of Saccharomyces bayanus var. uvarum, (for example S. bayanus MUCL 55125 (deposited at BCCM / MUCL Louvain-La-Neuve) or MUCL 31491 (obtained from BCCM / MUCL Louvain-La-Neuve), Saccharomyces bayanus var bayanus (for example, S. bayanus MUCL 31495 (obtained from BCCM / MUCL Louvain-La-Neuve), Saccharomyces bayanus BC S103 (obtained from Fermentis, Lesaffre group), Saccharomyces bayanus VR 44 (obtained from Fermentis, Lesaffre group), Saccharomyces cerevisiae (for example S. cerevisiae w-34/70 (obtained from Fermentis, Lesaffre group), Saccharomyces boulardii (obtained from Enterol®, biocodex gamma), and Kluyveromyces lactis var.drosophylarum, (eg Kluyveromyces lactis var.drosophylarum CBS 2103 ( obtained from the CBS-KNAW Fungal Biodiversity Center, Utrecht, the Netherlands) In a preferred embodiment, the yeast is Saccharomyces bayanus, preferably Saccharomyces bayanus varvarum or Saccharomyces bayanus bayanus, preferably Saccharomyces bayanus var. uva rum MUCL 55125 (filed with BCCM / MUCL Louvain-La-Neuve); Saccharomyces bayanus var. bayanus MUCL 31495 (obtained from BCCM / MUCL Louvain-La-Neuve), or Saccharomyces bayanus var. uvarum MUCL 31491 (obtained from BCCM / MUCL Louvain-La-Neuve), most preferably Saccharomyces bayanus var. uvarum MUCL 55125 (deposited at the BCCM / MUCL Louvain-La-Neuve). In particular, Saccharomyces bayanus appears to be very versatile with respect to incubation conditions, because it shows very good performance under a wide range of conditions.
In some embodiments, in the methods as herein described, the composition comprising fructan and sucrose, preferably pinulin, and sucrose, further comprises one or more additional free sugars (including sucrose), defined as described herein. above in an amount of at least 1% by weight, preferably at least 3% by weight, based on the total weight of the dry matter of the composition, and for example at most 70% by weight of free sugars ( including sucrose). More preferably at least 1% by weight and at most 60% by weight of free sugars (including sucrose), on a dry matter basis, more preferably at least 3% by weight and at most 50% by weight, based on of the material, dry. In the present context, the term "on the dry basis" refers to the% by weight of a respective component on the dry matter content of the composition (eg 1% by weight on a dry basis refers to 1 g per 100 g of dry matter).
The total dry matter can be determined gravimetrically as the residue remaining after drying. Typically, moisture is evaporated from the sample by oven drying. Typically, 5 g of sample are weighed into a previously weighed dry aluminum cup (Ohaus precision balance, capacity 410 g, sensitivity 0.001 g). The sample is placed in an oven at 103 ° C until the residual weight remains constant (at least 24 hours). The sample is cooled in a desiccator for 1 h and then immediately weighed. The results are expressed in% (g of dry matter per 100 g of sample).
Dry matter (%) = (m3 - m1) / (m2 - m1) x 100 m1 = weight of the dry aluminum cup (in g) m2 = weight of the aluminum cup with the sample before drying (in g) m3 = weight of the aluminum cup with the sample after drying (in g)
Preferably, the compositions comprising fructan and sucrose comprise at least 1.5% by weight of fructan, preferably inulin, based on the total weight of the composition. Preferably, these compositions comprise at most 80% by weight of fructan, preferably inulin, based on the total weight of the composition. Preferably, these compositions comprise at least 1.5% by weight and at most 75% by weight of fructan, preferably inulin, as indicated above. Other preferred embodiments illustrating the amounts of free sugars on the basis of the dry weight of the composition, at the beginning of the incubation step, in particular fructose, glucose and sucrose which may be present in the compositions comprising fructan and sucrose as presently described are described in Table 1 below.
Table 1
In one embodiment, in the compositions comprising fructan and sucrose, preferably inulin and sucrose, as presently described, the weight ratio based on the dry weight of sucrose and other free sugars, at the beginning of the incubation, preferably the ratio by weight on the basis of the dry weight of sucrose and one or more of fructose, and glucose, preferably all, to fructan, preferably inulin, is at least 1: 100 and at most 2.3: 1, more preferably at least 1:50 and at most 2: 1. In other embodiments, in the compositions comprising fructan and sucrose, preferably inulin, and sucrose as presently described, the weight ratio of free sugars, preferably the embodiments as described in Table 1, fructan, preferably inulin, is at least 1:10 and at most 1.5: 1, more preferably at least 1: 5 and at most 1: 1.
In certain optional embodiments, before and / or during step (b), a nitrogen source may be added to said composition comprising fructan and sucrose, preferably inulin and sucrose. The nitrogen source may be a source of organic (e.g., peptone) and / or inorganic (e.g., nitrate) nitrogen. In one embodiment, the nitrogen source may be provided in the form of a composition comprising other additives, such as additional nutrients, minerals, etc. In a preferred embodiment, the nitrogen source is extracted from yeast. In one embodiment, the amount of nitrogen source is at least 0.01% by weight, expressed as ammonium equivalent, and for example at most 1% by weight (based on the total weight of the composition), preferably at least 0.03% by weight and at most 1.0% by weight, more preferably at least 0.05% by weight and at most 1.0% by weight.
In certain optional embodiments, each of the methods as herein described may further comprise the step of aeration of the composition comprising fructan and sucrose, preferably inulin and sucrose, preferably after addition of the yeast as presently defined, at the beginning of the incubation, and / or during the incubation with the yeast as presently defined. It should be understood that the term "aeration" in the present context refers to a process by which an oxygen-containing gas, preferably air, is circulated in, mixed with or dissolved in the composition comprising fructan and sucrose, preferably inulin and sucrose, as defined herein. As a further, and unrestricted, recommendation, aeration may be accomplished by passing air through the liquid through a Venturi tube, venting turbines, or compressed air that can be combined with an air diffuser block (s), as well as fine bubble diffusers, large bubble diffusers or a linear aeration manifold. The preferred aeration rates are at least 0.01 wm and at most 1 wm (gas volume flow per unit volume of liquid per minute), preferably at least 0.05 wm and at most 1.0 wm. In one embodiment, when the yeast is Saccharomyces, such as Saccharomyces bayanus, no aeration is performed. In another embodiment, when the yeast is Kluyveromyces, such as Kluyveromyces lactis, aeration is performed.
In embodiments, each of the processes as herein described may further comprise the step of agitating the composition comprising fructan and sucrose, preferably inulin and sucrose, preferably after the addition of yeast as currently defined and / or during incubation with yeast as presently defined. It should be understood that the term "agitation" in the present context refers to a process by which the composition as defined herein is implemented, and therefore is mixed. By means of a further recommendation, agitation can be effected by shaking, stirring, rotating, or cyclic pumping of the liquid. For example, a magnetic stirrer or stirring bar may be used to effect agitation.
In other embodiments, each of the methods as herein described may further comprise the step of aeration and stirring the composition comprising fructan and sucrose, preferably inulin and sucrose, preferably after addition of the yeast as currently defined and / or during the incubation with the yeast as currently defined, the aeration and stirring being defined as defined above. It should be understood that aeration can include agitation and vice versa. For example, the introduction of air into the composition can set the composition in motion and thereby perform agitation. The other method, for example stirring by means of a helix can simultaneously introduce air into the composition.
In other embodiments, each of the methods as herein described may further comprise the step of removing the yeast after the incubation step, preferably after a specified time as currently defined elsewhere, such as, for example, also shown in Table 2. The yeast removal of the compositions after incubation as currently defined is known in the art. Without limitation, yeast removal can be performed by, for example, centrifugation, decantation, and / or filtration.
In some embodiments, in each of the methods as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the at least one yeast as currently defined at a temperature of above the freezing point of said composition, preferably at a temperature which is optimal for the respective yeast, preferably at a temperature of 10 ° C above or below the temperature which is optimal for the respective yeast. Optimum temperatures for yeasts as defined herein are known in the art. As a further recommendation, and without limitation, an optimal temperature as currently defined refers to the temperature at which growth is maximized. In a preferred embodiment, in each of the processes as described herein, yeast as defined herein is incubated with the composition comprising fructan and sucrose, preferably inulin and sucrose, at a temperature of from 0.degree. less -5 ° C. In a preferred embodiment, in each of the processes as described herein, yeast as defined herein is incubated with the composition comprising fructan and sucrose, preferably inulin and sucrose, at a temperature of from 0.degree. plus 40 ° C. In a preferred embodiment, in each of the processes as described herein, yeast as defined herein is incubated with the composition comprising fructan and sucrose, preferably inulin and sucrose, at a temperature of from 0.degree. less -5 ° C and not more than 40 ° C, more preferably at a temperature of at least 2 ° C and at most 35 ° C. In another preferred embodiment, the incubation is carried out at a temperature of at least 15 ° C and at most 35 ° C, for example at least 20 ° C and at most 30 ° C, for example 30 ° C or about 30 ° C. In another further preferred embodiment, the incubation is carried out at a temperature of at least -5 ° C and at most 15 ° C, for example at least 4 ° C and at most 10 ° vs.
In embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with at least one yeast as presently defined for at least 5 hours. for example for at least 10 hours, at least 15 hours; at least 50 hours, for example at least 75 hours; at least 4 days (i.e. 4x24 hours), for example at least 10 days; at least 30 days, for example at least 60 days, or at least 90 days, or at least 120 days. In embodiments, in each of the methods as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as presently defined for at most 12 months, by for example not more than 6 months, for example not more than 4 months, for example not more than 180 days, for example not more than 150 days, for example not more than 30 days (that is to say, 30 x 24 hours), for example not more than 20 days; not more than 150 hours, not more than 125 hours; not more than 50 hours, for example not more than 30 hours, or not more than 25 hours.
In embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as presently defined for at least 5 hours and plus 12 months, preferably for not less than 5 hours and not more than 6 months, for example not less than 5 hours and not more than 4 months, for example not less than 10 hours and not more than 30 hours or not less than 15 hours and not more than 25 hours ; at least 50 hours and not more than 150 hours, for example, not less than 75 hours and not more than 125 hours; at least 4 days (ie 4x24 hours) and not more than 30 days (ie 30 x 24 hours), for example at least 10 and at most 20 days; at least 30 days and not more than 180 days, for example, at least 60 days and 150 days, or at least 90 days and not more than 180 days, or not less than 120 days and not more than 150 days.
Preferred combinations of temperature and incubation time of the compositions comprising fructan and sucrose, preferably inulin and sucrose, with the at least one yeast as presently described are illustrated as embodiments in Table 2.
Table 2
In other embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined for a time sufficient for reduce one or more of the initial total weight or the. concentration (on a dry weight basis) of free sugars (including sucrose) of at least 10%. This reduction can be achieved by using one or more incubation steps. Preferably, said composition comprising fructan and sucrose is incubated with said at least one yeast until a reduction of at least 20% of the initial weight of free sugars (comprising sucrose) in said composition is obtained, for example at least 30%, for example at least 40%, for example at least 50%; for example at least 60%, for example at least 70%, for example at least 80%, preferably at least 90%, preferably at least 95%, for example at least 98%, for example a reduction of at least 99%. In other embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined for a time sufficient for reducing the fructose concentration (based on the dry weight) by at least 10%, for example at least 20%, for example at least 30%, for example at least 40%, for example at least 50%; for example at least 60%, for example at least 70%, for example at least 80%, preferably at least 90%, preferably at least 95%, for example at least 98%, for example a reduction of at least 99%. In other embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined for a time sufficient for reducing the glucose concentration (based on the dry weight) by at least 10%, for example at least 20%, for example at least 30%, for example at least 40%, for example at least 50%; for example at least 60%, for example at least 70%, for example at least 80%, preferably at least 90%, preferably at least 95%, for example at least 98%, for example a reduction of at least 99%. In other embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined for a time sufficient to reducing the sucrose concentration (on a dry weight basis) by at least 10%, for example at least 20%, for example at least 30%, for example at least 40%, for example at least 50%; for example at least 60%, for example at least 70%, for example at least 80%, preferably at least 90%, preferably at least 95%, for example at least 98%, for example a reduction of at least 99%. In other embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined for a time sufficient for reduce the combined concentration of fructose and glucose (based on dry weight) by at least 10%, for example at least 20%, for example at least 30%, for example at least 40%, for example at least 50% % for example at least 60%, for example at least 70%, for example at least 80%, preferably at least 90%, preferably at least 95%, for example at least 98%, for example a reduction of at least 99% by weight. In other embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined for a time sufficient for reduce the combined concentration of fructose and sucrose (on a dry weight basis) by at least 10%, for example at least 20%, for example at least 30%, for example at least 40%, for example at least 50% % for example at least 60%, for example at least 70%, for example at least 80%, preferably at least 90%, preferably at least 95%, for example at least 98%, for example a reduction of at least 99%. In other embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined for a time sufficient for reduce the combined glucose and sucrose concentration (on a dry weight basis) by at least 10%, for example at least 20%, for example at least 30%, for example at least 40%, for example at least 50%; % for example at least 60%, for example at least 70%, for example at least 80%, preferably at least 90%, preferably at least 95%, for example at least 98%, for example a reduction of at least 99%. In other embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined for a time sufficient for reduce the combined concentration of fructose, glucose and sucrose (on a dry weight basis) by at least 10%, for example at least 20%, for example at least 30%, for example at least 40%, for example at least 50%; for example at least 60%, for example at least 70%, for example at least 80%, preferably at least 90%, preferably at least 95%, for example at least 98%, for example a reduction of at least 99%. The times required to reach the defined free sugar (including sucrose) concentrations can be determined empirically, as is known in the art.
In embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined at a pH of at least 2.5, preferably at least 3.0. In embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, the yeast as defined herein is incubated with the composition comprising fructan and sucrose, preferably inulin and sucrose; at a pH of at least 2.5, for example at least 3.0, for example at least 3.5, for example at least 4.0, for example at least 5.0. In embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined at a pH of at most 8.5, preferably at most 7.5. In embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, the yeast as defined herein is incubated with the composition comprising fructan and sucrose, preferably inulin and sucrose, at a pH of at most 8.0, for example at most 7.5, for example at most 7.0, for example at most 6.5, for example at most 6.0. In embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined at a pH of at least 2.5 and at most 8.0, preferably at least 3.0 and at most 7.5. In embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, the yeast as defined herein is incubated with the composition comprising fructan and sucrose, preferably inulin and sucrose, at a pH of at least 4 and at most 7.0, for example at least 4.5 and at most 6.0, for example at least 5 and at most 7 , 0, for example at least 5.5 and at most 7.0. The pH can be defined and maintained as is known in the art.
In embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with yeast as presently defined, at least 103 CFU of yeast. being added at the beginning of the incubation per ml of composition comprising fructan and sucrose, preferably inulin and sucrose. In embodiments, in each of the methods as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as presently defined, at most 101 CFU of yeast being added at the beginning of the incubation per ml of composition comprising fructan and sucrose, preferably inulin and sucrose. In embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined, at least 103 CFU and plus 1010 colony-forming units CFU of yeast being added at the beginning of the incubation per ml of composition comprising fructan and sucrose, preferably inulin and sucrose. Colony forming units are known in the art and may, for example, be determined by plate counting. For example, it may be added to the compositions as defined herein, at least 103 CFU / ml and at most 109 CFU / ml, for example at least 104 CFU / ml and at most 109 CFU / ml, for example at least 105 CFU / ml. and at most 109 CFU / ml, for example at least 104 CFU / ml and at most 108 CFU / ml, for example at least 104 CFU / ml and at most 109 CFU / ml, for example at least 105 CFU / ml and not more than 108 CFU / ml. Advantageously, the above concentrations of yeast can be combined with the specific time and temperature embodiments as described in Table 2, or the specific times or temperatures as previously described.
In embodiments, in each of the processes as herein described, the composition comprising fructan and sucrose, preferably inulin and sucrose, is incubated with the yeast as currently defined, the yeast being supplied in the form of yeast lysates or yeast extract, such as protein or enzyme extract. It should be understood that to determine the amount of such lysates or extract, the corresponding amounts in CFU / ml as described above should be incubated with the compositions.
In a most preferred embodiment, in each of the methods as herein described, the composition is a liquid composition comprising fructan and sucrose, preferably inulin and sucrose, which is incubated with Saccharomyces, preferably Saccharomyces. bayanus, preferably Saccharomyces bayanus var. uvarum, the composition comprising at least 1% by weight and at most 70% by weight (based on the dry matter) of sucrose and other free sugars (including sucrose), preferably at least 1% by weight and not more than 70% by weight (based on dry matter) of sucrose and one or more of fructose, and glucose, preferably a mixture of all, based on the total weight of the dry matter of the composition .
In another most preferred embodiment, in each of the processes as herein described, the composition is a liquid composition comprising fructan and sucrose, preferably inulin and sucrose, most preferably comprising at least 30% by weight of fructan, preferably inulin, based on the total dry weight of the composition, and preferably comprising at least 1% by weight and at most 75% by weight (based on the material dry) of sucrose and optionally other free sugars (including sucrose), more preferably comprising at least 1% by weight and at most 75% by weight (based on the dry matter) of sucrose and one or more of fructose and glucose, preferably a mixture of all. Preferably, the composition is incubated with Saccharomyces bayanus, preferably Saccharomyces bayanus var. uvarum at a temperature of at least -5 ° C and at most 40 ° C, preferably at a temperature of at least 0.0 ° C and at most 35 ° C.
In another most preferred embodiment, in each of the processes as herein described, the composition is a liquid composition comprising fructan and sucrose, preferably inulin and sucrose, most preferably at least 30. % by weight and at most 99% by weight of fructan, preferably inulin, based on the total dry weight of the composition, said composition being incubated with Saccharomyces bayanus, preferably Saccharomyces bayanus var. uvarum, the composition comprising at least 1% by weight and at most 75% by weight (based on the dry matter) of sucrose and optionally other free sugars, preferably sucrose and one or more of the fructose and glucose, preferably a mixture of all, said composition being incubated at a temperature of at least -5 ° C and at most 40 ° C, preferably at a temperature of at least 0.0 ° C and at most ° C.
In one aspect, the invention further relates to a composition comprising fructan and sucrose, preferably inulin and sucrose and at least one yeast selected from the group consisting of or consisting of Saccharomyces bayanus (preferably Saccharomyces bayanus var. uvarum), Kluyveromyces lactis (preferably Kluyveromyces lactis var., drosophylarum, most preferably Kluyveromyces lactis var., drosophylarum CBS 2103 (obtained from the CBS-KNAW Center for Biodiversity, Utrecht, the Netherlands) In one embodiment Preferably, the yeast is Saccharomyces bayanus, preferably Saccharomyces bayanus varvarum.The embodiments described above with regard to the fructan compositions (in particular as regards the type, the quantity, the origin, the composition, DP, as well as embodiments related to free sugars, their types, and quantities) also apply to this aspect.
In another aspect, the invention relates to a yeast deposited in the Belgian Coordinated Collections of Microorganisms (BCCM) with the accession number MUCL 55125. It should be understood that this yeast is most preferred in the compositions, processes and uses according to the invention as currently described elsewhere.
The present invention further comprises a composition comprising fructan, sucrose and at least one yeast selected from the group consisting of Saccharomyces bayanus, Saccharomyces cerevisiae, and Kluyveromyces lactis, said composition comprising at least 30% by weight of fructan based on total weight of dry matter of said composition.
In yet another aspect, the invention relates to the use of yeast for removing, reducing, or removing sugars, preferably free sugars, more preferably carbohydrate monomers and / or carbohydrate dimers, preferably between all hexose and / or pentose monomers or dimers in a composition comprising fructan and sucrose, preferably inulin and sucrose, said yeast being selected from the group consisting of or consisting of Saccharomyces and Kluyveromyces; preferably selected from the group consisting of or consisting of Saccharomyces bayanus; Saccharomyces cerevisiae; Kluyveromyces lactis; Saccharomyces boulardii; still more preferably Saccharomyces bayanus var. uvarum, (eg Saccharomyces bayanus MUCL 55125 (deposited at BCCM / MUCL
Louvain-La-Neuve) or MUCL 31491 (obtained from BCCM / MUCL Louvain-La-Neuve), Saccharomyces bayanus var. bayanus (for example, S. bayanus MUCL 31495 (obtained from BCCM / MUCL Louvain-La-Neuve), Saccharomyces cerevisiae (for example S. cerevisiae w-34/70 (obtained from Fermentis, Lesaffre group), Saccharomyces boulardii ( obtained from Enterol®, biocodex gamma) and Kluyveromyces lactis var. drosophylarum, (most preferably Kluyveromyces lactis var., drosophylarum CBS 2103 (obtained from the CBS-KNAW Center for Biodiversity, Utrecht, The Netherlands); more preferably Saccharomyces bayanus var uvarum, (for example Saccharomyces bayanus MUCL 55125 (deposited at BCCM / MUCL Louvain-La-Neuve) or MUCL 31491 (obtained in BCCM / MUCL Louvain-La-Neuve), Saccharomyces bayanus var. for example, S. bayanus MUCL 31495 (obtained in BCCM / MUCL Louvain-La-Neuve), Saccharomyces cerevisiae (for example S. cerevisiae w-34/70 (obtained from Fermentis, Lesaffre group), Saccharomyces boulardii (obtained from Enterol®, biocodex gamma) ably Saccharomyces bayanus var. uvarum MUCL 55125 (deposited at the BCCM / MUCL Louvain-La-Neuve). Preferably, the free sugars are selected from one or more of fructose, glucose, and sucrose, preferably all. The embodiments described above with regard to the fructan compositions (in particular as regards the type, the quantity, the origin, the composition, the DP (average), as well as the embodiments associated with free sugars , their types, and quantities), as well as the incubation time and temperature also apply to compositions of this aspect.
The present invention further comprises the use of a yeast selected from the group consisting of Saccharomyces and Kluyveromyces for reducing the amount of sucrose in a composition comprising sucrose and at least 30% by weight of fructan based on the total weight of dry matter of said composition.
Aspects and embodiments of the invention are further illustrated by the following non-limiting examples.
EXAMPLES
protocols
Measurement of dry matter
The total dry matter is determined gravimetrically as the residue remaining after drying. The moisture is evaporated from a sample by drying in an oven. 5 g of sample are weighed in a previously weighed dry aluminum cup (Ohaus precision balance, capacity 410 g, sensitivity 0.001 g). The sample is placed in an oven at 103 ° C until the residual weight remains constant (at least 24 hours). The sample is cooled in a desiccator for 1 h and then immediately weighed. The results are expressed in% (g of dry matter per 100 g of sample).
Dry matter (%) = (m3 - m1) / (m2 - m1) x 100 m1 = weight of the dry aluminum cup (in g) m2 = weight of the aluminum cup with the sample before drying (in g) m3 = weight of the aluminum cup with the sample after drying (in g) Determination of the molecular weight distribution of the inulin
Determination of the molecular weight distribution of the fructan sample is performed by high performance anion exchange chromatography coupled with pulsed amperometric detection (HPAEC-PAD) on a Thermo Scientific - Dionex ICS 5000 chromatography system. Separation of the different chain lengths is carried out with a Carbopac PA100 4 mm x 250 mm column (+ precolumn) at 40 ° C. with a flow rate of 1 ml / min. 160 mM sodium hydroxide is used as the eluent. A sodium acetate gradient during the test makes it possible to separate the different lengths of the chain. The software makes it possible to determine each corresponding peak area in nC * min. Determination of the average Dp in number
Different concentrations of a standard inulin are injected to assign the peaks in the chromatogram based on the standard retention time and to plot the calibration curves.
The calibration curves make it possible to determine the mass concentration of each molecular species of inulin in the sample Ci. The molar concentration (Ni) of the molecules with residues is calculated as Ci / MWi, where MWi is the weight. Molecular molecules with residues.
The number-average degree of polymerization Dpn is calculated by - Σ, ιν, Ρρι where Dpi is the number of residues. Determination of free sugars
In a weighed bottle (Schott), about 5 g of a representative sample (m4 at 0.001 g) are accurately weighed. Then, about 10 g of phosphate buffer (0.1 M) at pH = 7.0 are added and the sample is heated at 80 ° C for 15 minutes in a water bath. Then, the sample is cooled to room temperature and the total weight of the solution is brought to 40 g with deionized water (m5 to 0.001 g).
The first dilution factor is D1 = m5 / m4.
Finally, appropriate dilutions (D2) for HPAEC-PAD assays with appropriate calibration (glucose, fructose, sucrose) are performed.
The amounts of free glucose, free fructose and free sucrose are determined by multiplying the results of HPAEC-PAD by D1 * D2 and are expressed in g / kg of the sample or in% by weight on the basis of dry matter . Determination of the amount of inulin
Principle
The amount of inulin is determined from the amount of glucose and fructose released by enzymatic hydrolysis. Glucose, fructose and free sucrose are initially determined on a representative unhydrolyzed sample. Then, enzymatic hydrolysis is performed and glucose and total fructose are determined. The released amounts are obtained by difference taking into account the amounts of glucose and fructose released from sucrose.
The process is based on the AOAC997.08 process with slight adaptations as described below. Determination of free sugars
The amounts of free glucose (Gf), free fructose (Ff) and free sucrose (S) are determined by HPAEC-PAD as described above.
Enzymatic hydrolysis - determination of total fructose and total glucose
In a weighed beaker, about 1 g of a representative sample (m6 at 0.001 g) is accurately weighed. Then about 20 g of acetate buffer (0.1 M) at pH 4.75 is added and the mixture is homogenized. Then, the sample is heated at 80 ° C for 15 minutes in a water bath and cooled to 60 ° C in a water bath (allowed to equilibrate). Then, 50 μl of Fructozyme (Novozym SP 230®, Novo Nordisk) are added and the mixture is homogenized. Then, the vial is closed and the mixture is incubated in a water bath at 60 ° C for 2 hours. The sample is cooled to room temperature and the mass of the solution is brought to 40 g with deionized water (m 7 to 0.001 g). Finally, the sample is homogenized.
The first dilution factor is D3 = m7 / m6
Appropriate dilutions (D4) for HPAEC-PAD assays with appropriate calibration (glucose and fructose) are performed.
The amounts of total glucose (Gt) and total fructose (Ft) are determined by multiplying the results of HPAEC-PAD by D3 * D4 and are expressed in g / kg of the initial composition.
calculations
The glucose released from the inulin fraction is Gi = Gt-Gf-S / 1.9 (in g / kg)
Fructose released from the inulin fraction is Fi = Ft-Ff-S / 1.9 (in g / kg)
The amount of inulin in the sample is k (Gi + Fi) where k is a factor taking into account the increase in dry matter due to the hydrolysis of inulin. In our examples, k is set to 0.91.
Loss of inulin
Inulin loss is defined as the difference between the amount of inulin before and after incubation with yeast expressed as a mass percentage of the initial amount. Determination of organic acids
Determination of the organic acid concentration is performed by a high performance liquid chromatography system (LCM1 Waters) comprising a UV detector (Waters 2487), an automatic injector (Waters 717) and a controller (Waters 600). Peak separation is performed by a HPX-87H Biorad column at 65 ° C with a flow rate of 0.8 ml / min. H2S04 0.0045 N is used as the eluent.
The calibration line is obtained by injection of 10 μl, 25 μΙ, 40 μΙ, 50 μΙ of a stock solution of 1 g / l of different acids to be assayed. The calibration curves allow the determination of the concentration of each molecular species in the sample. 25 μΙ of the sample are injected for a duration of analysis of 20 minutes. Determination of alcohols and volatile components
The determination of the alcohols and volatile components is carried out by gas chromatography coupled to an FID detector on a Perkin-Elmer 8000 chromatographic system. Peak separation is performed with a CP WAX-52 column. The analysis is performed using the headspace technique. The gas phase in equilibrium with the liquid phase is injected in gas chromatography according to a temperature program (preheating: 60 ° C / 20 min, heating: increase of 60 ° per minute up to 110 °, injector temperature (HS40 Perkin -Elmer): 110 ° C, FID detector temperature: 250 ° C).
Mixtures of volatile component standards at different concentrations are injected to plot the calibration curves and assign the peaks in the chromatogram based on the retention time of the standard. The calibration curves make it possible to determine the concentration of each molecular species in the sample.
Preparation of compositions comprising fructan and sucrose (inulin-rich extract) - step (a)
A1-A3 Compositions
Chicory roots are washed and cut into chips. Countercurrent diffusion with hot water (70 ° C) is then used to extract inulin from the chips. The ratio of chips to water is 1. The extraction time is 2 hours. The resulting juice containing inulin in solution is roughly filtered to remove spent chips. The resulting juice is then filtered to remove the small insoluble material. The pH is adjusted to 4 with 25% HCl. A concentration step at 100 ° C. for 1 hour makes it possible to increase the dry matter to 40% w / w, so as to prepare A1-A3 compositions.
Composition F
Chicory roots are washed and cut into chips. Countercurrent diffusion with hot water (70 ° C) is then used to extract inulin from the chips. The ratio of chips to water is 1. The extraction time is 2 hours. The resulting juice containing inulin in solution is roughly filtered to remove spent chips. The resulting juice is then filtered to remove the small insoluble material. A concentration step at 100 ° C. for 1 hour makes it possible to increase the dry matter to 40% w / w, so as to prepare the composition F.
Preparation of compositions comprising fructan and sucrose (inulin-rich compositions) - step (a)
Compositions B1-B7 217 g of Fibruline® Instant (marketed by Cosucra Groupe Warcoing) are suspended in 1 kg of phosphate buffer solution pH 5.8.
The phosphate buffer is prepared as follows: 467.5 ml of 0.2 mol / l solution of KH2PO4 and 32.5 ml of 0.2 mol / l K2HPO4 are mixed. Then, the mixture is brought to 1 I with distilled water. 90 g of Fibrulin® Instant suspension are placed in a 250 ml vial and sterilized (20 min, 121 ° C) to prepare B1-B7 compositions.
Composition C 434 g of Fribuline® Instant (marketed by Cosucra Groupe Warcoing) are suspended in 2 kg of demineralized water. Then, 1800 g of the Fibruline® Instant suspension is added to the 2 I bioreactor and sterilized (20 min, 121 ° C) to prepare the C composition.
Composition D 31.5 kg of Fribulose® F90 (marketed by Cosucra Groupe Warcoing) are suspended in 40 kg of demineralized water. Then, 40 kg of the Fibrulose® F90 suspension is added to a 60 I drum, so as to prepare the composition D. The composition D is not sterilized.
Composition E 10.5 kg of Fribulose® F90 (marketed by Cosucra Groupe Warcoing) are suspended in 40 kg of demineralized water. Then, 40 kg of the Fibrulose® F90 suspension is added to the 60 I drum, so as to prepare the composition E. The composition E is not sterilized.
The concentrations of inulin and free sugars of compositions A-F are shown in Table 3.
Table 3
Example 1: Specificity of different yeasts for a composition comprising inulin, sucrose and other free sugars - step (b).
An extract of yeast solution is prepared as follows: 10 g of yeast extract (Merck) are dissolved in 100 ml of demineralized water. Then, the solution is sterilized (20 min, 121 ° C). 10 g of the sterilized yeast extract solution are added to 90 g of composition B1-B5 respectively. No additional ventilation is applied.
The compositions B1-B5, supplemented with yeast extract, are inoculated at a concentration of 105 CFU / ml with different yeasts, respectively: Saccharomyces cerevisiae w-34/70 (from Fermentis, Lesaffre group), Kluyveromyces lactis CBS 2103 ( of the CBS-KNAW Fungal Biodiversity Center, Utrecht, the Netherlands), Saccharomyces bayanus var. bayanus MUCL 31495 (from MUCL Louvain-La-Neuve, Belgium), Saccharomyces bayanus var. uvarum MUCL 31491 (from MUCL Louvain-La-Neuve, Belgium) and Saccharomyces bayanus var. uvarum MUCL 55125 (deposited in the Belgian Coordinated Collections of Microorganisms (BCCM) under the order number MUCL 55125). The compositions are incubated at different temperatures (20 ° C. and 30 ° C.) with stirring at a speed of 160 r / min. The results of these tests are shown in Figures 1 to 15.
Figures 1, 2, 3 and 4 show growth (measured by optical density at 660 nm) over time at 30 ° C of Saccharomyces cerevisiae w-34/70, Kluyveromyces lactis CBS 2103, Saccharomyces bayanus var. bayanus MUCL 31495, and Saccharomyces bayanus var. uvarum MUCL 31491, respectively, incubated with compositions B1-B4 respectively in a 250 ml flask.
Figure 5 shows growth (measured by optical density at 660 nm) over time at 30 ° C and 20 ° C of Saccharomyces bayanus var. uvarum MUCL 55125 incubated with the composition B5 in the 250 ml flask.
FIGS. 6, 8, 10, 12 and 14 represent the evolution of the concentration of free sugars over time at 30 ° C. of compositions B1-B5 respectively incubated with Saccharomyces cerevisiae w-34/70, Kluyveromyces lactis CBS 2103, Saccharomyces bayanus var. bayanus MUCL 31495, Saccharomyces bayanus var. uvarum MUCL 31491 and Saccharomyces bayanus var. uvarum MUCL 55125 respectively in a 250 ml flask. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w based on the total dry matter).
Figures 7, 9, 11, 13 and 15 show the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of the compositions B1 -B5 respectively incubated with Saccharomyces cerevisiae w-34/70, Kluyveromyces lactis CBS 2103, Saccharomyces bayanus var. bayanus MUCL 31495, Saccharomyces bayanus var. uvarum MUCL 31491 and Saccharomyces bayanus var. uvarum MUCL 55125 respectively in a 250 ml flask. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition). From Figures 6 to 15, it is clear that Saccharomyces and Kluyveromyces are effective for degrading free sugars. From the figures, it can be observed that Saccharomyces bayanus var. uvarum MUCL 55125 degrades free sugars more rapidly than Saccharomyces cerevisiae w-34/70, Kluyveromyces lactis CBS 2103, Saccharomyces bayanus var. bayanus MUCL 31495 and Saccharomyces bayanus var. uvarum MUCL 31491, and / or has a higher specificity for free sugars than for inulin.
Nonlimiting examples of optimal conditions are presented in Table 4. These conditions allow the degradation of a quantity of free sugars and a limited degradation of the inulin.
Table 4
Example 2: Incubation of a Composition Comprising Inulin and Sucrose with Saccharomyces bayanus var. uvarum MUCL 55125 - (step b)
Example in a bioreactor of 2 I
A solution of yeast extract is prepared as follows: 20 g of yeast extract (Merck) are dissolved in 200 ml of deionized water and sterilized (20 min, 121 ° C). 200 g of the sterilized yeast extract solution is added to a 2 I bioreactor containing 1800 g of the composition C.
No additional ventilation is performed. The pH of the composition is maintained at a value of 5 by means of a peristaltic pump dispensing a basic solution containing 10 mol / l sodium hydroxide and an acidic solution containing 30% vol / phosphoric acid. flight.
Composition C, supplemented with yeast extract, is incubated at a concentration of 105 CFU / ml at 30 ° C. with stirring at a speed of 160 r / min with Saccharomyces bayanus var. uvarum MUCL 55125.
After 62 h, the loss of inulin is less than 2%.
Figure 16 shows growth (measured by optical density) over time at 30 ° C of Saccharomyces bayanus var. uvarum MUCL 55125 incubated with the composition C in a bioreactor of 2 I.
FIG. 17 represents the evolution of the concentration of free sugars over time at 30 ° C. of the composition C incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in a bioreactor of 21. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w based on the total dry matter).
Figure 18 shows the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of composition C incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in a bioreactor of 2 I. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) - normalized according to the dilution of the composition).
Example in a barrel of 60 I 40 g of yeast extract (Merck) are dissolved in 100 ml of demineralized water. Then, the solution is sterilized (20 min, 121 ° C) and added to 40 kg of composition D.
No additional ventilation is performed. The pH is not regulated.
Composition D, supplemented with yeast extract, is inoculated at a concentration of 105 CFU / ml at 20 ° C. with shaking with Saccharomyces bayanus var. uvarum MUCL 55125.
FIG. 19 represents the evolution of the concentration of free sugars over time at 20 ° C. of composition D incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in a drum of 60 I. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w on the basis of the total dry matter).
Figure 20 shows the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 20 ° C of composition D incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in a 60 l drum. The analyzes are performed by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition).
After 150 hours, the loss of inulin for composition D is less than 2%. 40 g of yeast extract (Merck) are dissolved in 100 ml of demineralized water. Then, the solution is sterilized (20 min, 121 ° C) and added to 40 kg of composition E.
No additional ventilation is performed. The pH is not regulated.
Composition E, supplemented with yeast extract, is inoculated at a concentration of 105 CFU / ml at 20 ° C. without stirring with Saccharomyces bayanus var. uvarum MUCL 55125.
FIG. 21 represents the evolution of the concentration of free sugars over time at 20 ° C. of composition E incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in a drum of 60 I. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w).
Figure 22 shows the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 20 ° C of composition E incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in a 60 l drum. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (n) * retention time (min) -normalized according to the dilution of the composition).
After 50 h, the loss of inulin for composition E is less than 4%.
Example 3 Incubation of a Composition Comprising Inulin and Sucrose with Saccharomyces bayanus var. uvarum (MUCL 55125) - step (b) 2 kg of composition A1 are inoculated at a concentration of 105 CFU / ml with Saccharomyces bayanus var. uvarum MUCL 55125 and incubated at different temperatures (20 ° C and 30 ° C) with a stirring speed of 160 r / min in a bioreactor of 2 I.
The composition A1 is not sterilized. No additional nitrogen sources and aeration are added. The pH of the composition is maintained at a value of 5 by means of a peristaltic pump dispensing a basic solution containing 10 mol / l sodium hydroxide and an acidic solution containing 30% vol / phosphoric acid. flight.
Figure 23 shows growth at 30 ° C (measured by optical density) over time of Saccharomyces bayanus var. uvarum MUCL 55125 incubated with the composition A1 in a bioreactor of 2 I.
Figure 24 shows the evolution of free sugar concentration over time at 20 ° C (A) and at 30 ° C (B) of the A1 composition incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in bioreactor of 2 I. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w on the basis of the total dry matter).
Figure 25 shows the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 20 ° C (A) and 30 ° C (B). the A1 composition incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in a bioreactor of 2 I. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) - normalized according to the dilution of the composition).
Table 5 shows the list and concentration of by-products isolated from composition A1 before and after incubation with Saccharomyces bayanus var. uvarum MUCL 55125 at 20 ° C.
Table 5
It can be seen from the results of Table 5 that incubation with Saccharomyces bayanus var. uvarum MUCL 55125 produces mainly ethanol as a degradation product of free sugars.
Example 4 Incubation of a Composition Comprising Inulin and Sucrose with Saccharomyces bayanus var. uvarum MUCL 55125, at 4 ° C - step (b) 1 kg of composition A2 is inoculated at a concentration of 105 CFU / ml with Saccharomyces bayanus var. uvarum MUCL 55125 and incubated at 4 ° C in a flask of 1 I with a stirring speed of 110 r / min. Composition A2 is not sterilized. No additional nitrogen sources and aeration are added. The pH is not controlled.
FIG. 26 represents the evolution of the concentration of free sugars over time at 4 ° C. of the composition A2 incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in a vial of 1 I. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w on the basis of the total dry matter).
Figure 27 shows the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 4 ° C of the A2 composition incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in a flask of 1 I. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition). Even at low temperatures, Saccharomyces bayanus var. uvarum MUCL 55125 is active to perform adequate degradation of free sugars (but more slowly) without significant degradation of inulin, compared to room temperature tests (Figures 24 and 25).
Example 5: Study of the metabolism of Saccharomyces bayanus var. uvarum MUCL 55125 with aeration - step (b) 2 kg of composition A3 are inoculated with Saccharomyces bayanus var. uvarum MUCL 55125 at a concentration of 105 CFU / ml and incubated at 25 ° C in a bioreactor of 2 I with a stirring speed of 160 r / min and an aeration rate of 1 l / min.
Composition A3 is not sterilized. No additional nitrogen source is added. The pH of the composition A3 is maintained at a value of 5 by means of a peristaltic pump dispensing a basic solution in the form of 10 mol / l sodium hydroxide and an acidic solution in the form of 30% vol / vol phosphoric acid. .
Figure 28 shows the evolution of the concentration of free sugars over time at 25 ° C with aeration of the composition A3 incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in a bioreactor of 2 I. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w on the basis of the total dry matter).
Figure 29 shows the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 25 ° C with aeration of the A3 composition incubated with Saccharomyces bayanus var . uvarum MUCL 55125 in a bioreactor of 2 I. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) - normalized according to the dilution of the composition). Aeration stimulates the fermentation of free sugars. Indeed, the degradation of free sugars by yeast is much faster compared to fermentation that occurs without aeration (Figures 24 and 25).
Example 6 Comparative Incubation of a Composition Comprising Inulin and Sucrose with Rhodotorula dairenensis CBS 7294 (step b)
A solution of yeast extract is prepared as follows: 10 g of yeast extract (Merck) are dissolved in 100 ml of demineralized water. Then, the solution is sterilized (20 min, 121 ° C). 10 g of the sterilized yeast extract solution are added to 90 g of composition B6. No additional ventilation is performed.
The composition B6, supplemented with yeast extract, is inoculated with Rhodotorula dairenensis CBS 7294 (from the CBS-KNAW fungal biodiversity center, Utrecht, The Netherlands) at a concentration of 105 CFU / ml and incubated at 30 ° C. with a stirring speed of 160 r / min.
Figure 30 shows the evolution of the concentration of free sugars over time at 30 ° C of the B6 composition incubated with Rhodotolula dairenensis CBS 7294 in a 250 ml flask. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w based on the total dry matter).
Figure 31 shows the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of the B6 composition incubated with Rhodotolula dairenensis CBS 7294 in a 250 ml flask. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition). From Figures 30 to 31 and 17 to 18, it is clear that Saccharomyces bayanus var. uvarum MUCL 55125 degrades all the free sugars tested whereas Rhodotolula dairenensis CBS 7294 generates more free sugars. Saccharomyces bayanus var. uvarum MUCL 55125 clearly has a higher specificity for free sugars than inulin.
Example 7 Comparative Incubation of a Composition Comprising Inulin and Sucrose with Aureobasidium Pullulans CBS 621.80- (Step b)
A solution of yeast extract is prepared as follows: 10 g of yeast extract (Merck) are dissolved in 100 ml of demineralized water. Then, the solution is sterilized (20 min, 121 ° C). 10 g of the sterilized yeast extract solution are added to 90 g of composition B7. No additional ventilation is performed.
Composition B7, supplemented with yeast extract, is inoculated with Aureobasidium Pullulans CBS621.80 (obtained from the CBS-KNAW Fungal Biodiversity Center, Utrecht, the Netherlands) at an optical density (OD at 660 nm) of 0.1 and incubated at 30 ° C with a stirring speed of 160 r / min.
Figure 32 shows the evolution of the concentration of free sugars over time at 30 ° C of the composition B7 incubated with Aureobasidium Pullulans CBS 621.80. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w based on the total dry matter).
Figure 33 shows the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of the B7 composition incubated with Aureobasidium Pullulans CBS 621.80. The analyzes are carried out by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) - normalized according to the dilution of the composition). From Figures 32 to 33 and 17 to 18, it is clear that Saccharomyces bayanus var. uvarum (MUCL 55125) degrades all the free sugars tested while Aureobasidium Pullulans CBS 621.80 generates more free sugars. Saccharomyces bayanus var. uvarum MUCL 55125 clearly has a higher specificity for free sugars compared to inulin.
Example 8 Incubation of a Composition Comprising Inulin and Sucrose with Saccharomyces bayanus var. uvarum MUCL 55125 - (step b) 40 kg of composition F in a 60 l drum were inoculated at a concentration of 105 CFU / ml at 30 ° C. without stirring with Saccharomyces bayanus var. uvarum MUCL 55125.
No additional ventilation is performed. The pH is not regulated.
Figure 34 shows the evolution of the concentration of free sugars over time at 30 ° C of the composition F incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in a drum of 60 I. The analyzes are carried out by HPAEC-PAD (results expressed in% w / w on the basis of the total dry matter).
Figure 35 shows the evolution of the peak area of GF2, F2, GF3, F3, GF4, F4, GF5, F5 and GF6 over time at 30 ° C of the F composition incubated with Saccharomyces bayanus var. uvarum MUCL 55125 in a 60 l drum. The analyzes are performed by HPAEC-PAD (the area being expressed in nanocoulombs (nC) * retention time (min) -normalized according to the dilution of the composition).

权利要求:
Claims (15)
[1]
A process for treating a composition comprising fructan and sucrose, comprising the steps of (a) providing a composition comprising fructan and sucrose, wherein said composition comprising fructan and sucrose comprises at least 30% by weight (% w / w) fructan based on the total dry weight of said composition; and (b) incubating said composition comprising fructan and sucrose with at least one yeast selected from the group consisting of Saccharomyces and Kluyveromyces; until a reduction of at least 10% of the initial weight of sucrose in said composition is obtained.
[2]
The method of claim 1, wherein said at least one yeast is selected from the group consisting of Saccharomyces bayanus, Kluyveromyces lactis, Saccharomyces cerevisiae, and Saccharomyces boulardii.
[3]
The method of claim 1 or 2, wherein said composition comprising fructan and sucrose further comprises one or more free sugars, preferably wherein said free sugars are selected from the group consisting of glucose and fructose.
[4]
4. The process according to any one of claims 1 to 3, wherein said fructan has a number average degree of polymerization of at least 3.
[5]
5. Process according to any one of claims 1 to 4, wherein said fructan is of plant origin, preferably from chicory.
[6]
The process of any one of claims 1 to 5, wherein said fructan is inulin, preferably chicory inulin.
[7]
The method of any one of claims 1 to 6, wherein said yeast is Saccharomyces bayanus var. uvarum deposited in the Belgian Coordinated Collections of Microorganisms (BCCM) with serial number MUCL 55125.
[8]
The method of any one of claims 1 to 7, wherein said composition comprising fructan and sucrose is incubated with said yeast at a temperature of at least the freezing point of the composition.
[9]
The method of any one of claims 1 to 8, wherein said composition comprising fructan and sucrose is incubated with said yeast at a pH of at least 2.5.
[10]
The process according to any one of claims 1 to 9, wherein said composition comprising fructan and sucrose comprises at least 5% by weight and at most 80% by weight of dry matter on the basis of the total weight of the composition .
[11]
The process according to any one of claims 3 to 10, wherein the weight ratio of free sugars comprising fructan sucrose in said composition comprising fructan and sucrose at the beginning of the incubation is at least 1: 100.
[12]
The method according to any one of claims 1 to 11, wherein at the end of said incubation step, the fructan weight of said composition comprising fructan and sucrose is at most 20% less than the weight initial fructan at the beginning of said incubation, preferably at most 10%, most preferably at most 5%.
[13]
13. Composition comprising fructan, sucrose and at least one yeast selected from the group consisting of Saccharomyces bayanus, Saccharomyces cerevisiae, Kluyveromyces lactis, and Saccharomyces boulardii, said composition comprising at least 30% by weight (% w / w) of fructan on the basis of the total weight of dry matter of said composition.
[14]
14. Yeast deposited in the Belgian Coordinated Collections of Microorganisms (BCCM) with serial number MUCL 55125.
[15]
15. Use of a yeast selected from the group consisting of Saccharomyces and Kluyveromyces for reducing the amount of sucrose in a composition comprising sucrose and at least 30% by weight (% w / w) of fructan based on the total weight of dry matter of said composition.

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同族专利:

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公开号 | 公开日

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引用文献:

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公开号 | 申请日 | 公开日 | 申请人 | 专利标题

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WO2006108697A1|2005-04-15|2006-10-19|Bayer Cropscience Ag|Long-chain inulin|

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WO2008133512A1|2007-04-26|2008-11-06|Csm Nederland B.V.|Yeast-leavened dough and dry mix for preparing such a dough|

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法律状态:

优先权:

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申请号 | 申请日 | 专利标题

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EP14173660|2014-06-24|

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EP141736603|2014-06-24|PL15736212T| PL3161011T3|2014-06-24|2015-06-24|Methods and compositions for processing dietary fibers|

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EP15736212.0A| EP3161011B1|2014-06-24|2015-06-24|Methods and compositions for processing dietary fibers|

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CN201580033651.3A| CN106795478A|2014-06-24|2015-06-24|Method and composition for processing dietary fiber|

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ES15736212T| ES2727083T3|2014-06-24|2015-06-24|Methods and compositions for processing dietary fibers|

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PCT/EP2015/064215| WO2015197670A1|2014-06-24|2015-06-24|Methods and compositions for processing dietary fibers|

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US15/313,564| US10111455B2|2014-06-24|2015-06-24|Methods and compositions for processing dietary fibers|

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CL2016003259A| CL2016003259A1|2014-06-24|2016-12-20|Methods and compositions for the processing of dietary fibers|