PROCESS FOR EXTRACTING PEAS PROTEINS

专利摘要:
The present invention relates to a process for the extraction and purification of pea proteins. To this end, according to the invention, peas are subjected to fermentation, preferably by lactic acid bacteria, before being crushed. Preferably, the method of extracting pea proteins comprises the steps of: (a) subjecting to fermentation an aqueous composition comprising peas; (b) crushing said peas; (c) fractionating said ground peas in order to obtain at least one protein fraction; and (d) isolating pea proteins from said at least one protein moiety. The present invention also describes products intended for human or animal consumption comprising pea proteins obtained according to the invention.
公开号:BE1021189B1
申请号:E2014/0174
申请日:2014-03-13
公开日:2015-06-23
发明作者:Audrey Bourgeois；Eric Bosly；Frédéric Mansy；Julie Lebesgue
申请人:Sa Cosucra Groupe Warcoing；
IPC主号:

专利说明:

PROCESS FOR EXTRACTING PEP PROTEINS DOMAIN OF THE INVENTION
The present invention relates to methods for extracting and purifying proteins. In particular, the present invention relates to the extraction of pea proteins. The invention further relates to pea proteins obtainable by the above-mentioned methods, as well as to food and feed products containing pea proteins. The invention also relates to the use of pea protein in the food industry for food or feed.
BACKGROUND OF THE INVENTION
Protein isolates of plant origin represent an interesting alternative or complement to animal proteins in human and animal nutrition. For example in foodstuffs, the addition of vegetable proteins can effectively replace animal proteins, often at a lower cost. In addition, many products usually containing animal protein, especially dairy products, can be a major source of food allergies.
Legumes are remarkable because most of them have symbiotic nitrogen-fixing bacteria in structures called root nodules. This arrangement means that root nodules are nitrogen sources for legumes, making them relatively rich in vegetable protein. All proteins contain nitrogen amino acids. Nitrogen is therefore an essential ingredient for the production of proteins. Legumes are therefore the best sources of vegetable protein. In addition to being high in protein, legumes such as peas (Pisum sativum) are readily available and have a particularly well-balanced amino acid composition. As a result, they represent a source of protein which constitutes an interesting alternative to animal proteins.
The main difficulties in producing vegetable proteins revolve around the composition and purity of proteins, and include aspects relating to, for example, extraction, fractionation, and pre- and post-isolation treatments. Until plant proteins are isolated and available in a more or less pure form, all prior manipulations have a strong impact on the quality of isolated plant proteins. For example, the type and amount of impurities in isolates or protein extracts determine their final value. These impurities include, for example, carbohydrates. For example, legumes contain a significant proportion of so-called "flatulent" sugars (raffinose, stachyose, and verbascose), which are particularly undesirable. While carbohydrates are generally undesirable impurities in the final protein isolate, other impurities, such as vitamins or minerals, can not, by definition, be undesirable, or may even be nutritionally beneficial. or physicochemical of the protein isolate. In addition to having an impact on the final composition of the isolates or protein extracts, the extraction and / or purification process can have a dramatic effect on the physicochemical or functional properties of the protein isolate. In particular, the solubility, the viscosity, the emulsification capacity, the color, the taste, or the smell of the proteins are strongly influenced by the techniques used.
As the reading of the preceding paragraphs has shown, obtaining a high quality protein isolate with specific desired properties can be difficult, and often involves many expensive and / or time consuming manipulations. In view of this, it remains necessary to improve the isolation of plant proteins, particularly legumes, such as peas. Therefore, one of the objects of the present invention is to overcome or improve at least one of the disadvantages of the prior art, or to provide a useful alternative.
SUMMARY OF THE INVENTION
According to a first aspect, the present invention relates to a method for extracting pea proteins. The method of extracting pea proteins comprises the steps of: (a) subjecting an aqueous composition comprising peas to fermentation; (b) grinding said peas; (c) fractionating said ground peas to obtain at least one protein fraction; and (d) isolating the pea proteins from the at least one protein fraction.
According to the present invention, the extraction of pea proteins involves the fermentation of the peas before grinding them. During or after grinding, the pea proteins are separated and isolated. Downstream purification steps are also contemplated.
According to a second aspect, the present invention relates to pea proteins that can be obtained or are obtained using the method according to the first aspect of the invention.
According to a third aspect, the present invention relates to an edible composition, preferably a product for human or animal consumption, comprising pea proteins according to the second aspect of the invention, or pea proteins obtained using of the method according to the first aspect of the invention.
According to a fourth aspect, the present invention relates to the use of pea proteins according to the second aspect of the invention, or pea proteins obtained using the method according to the first aspect of the invention in products intended for human or animal food, preferably in dairy products, confectionery products, beverages, meat products, vegetarian products, dietary supplements, nutritional products for weight control and sports medical use, food for the elderly and in bakery products.
The present inventors have surprisingly found that the fermentation of peas (Pisum sativum) has a beneficial effect on several physicochemical parameters and related to the quality of extracts, concentrates, or isolates of proteins derived therefrom.
When whole peas are fermented before being ground, fermentation microorganisms, as well as fermentation byproducts, such as lactic acid, but also secreted compounds such as enzymes, which may have impact on the downstream treatment, are separated from the peas after fermentation easily and cost-effectively. Moreover, unexpectedly, during the fermentation of whole peas, the mono-, di-, and / or oligosaccharide contents of the peas, and in particular the contents of mono- and dimeric sugars, such as glucose, fructose sucrose, galactose, and / or flatulent sugar contents, such as raffinose, stachyose, and verbascose, all contained in peas, are considerably reduced, which is even more surprising when one considers the limited duration of fermentation in some embodiments. Without wishing to be bound by any theory, it is thought that fermentation, besides the consumption of sugars, accelerates the diffusion of sugars outside the peas. Advantageously reducing the amounts of mono-, di-, and oligosaccharides minimizes, for example, the consumption of water and energy in downstream processes, such as subsequent purification. Therefore this represents an economic advantage.
On the other hand, it has been advantageously found that fermentation until a specific level of hydration is achieved, and / or until a specified pH of peas is reached, as detailed below , leads to extracts, concentrates and isolates of pea proteins having particular physicochemical and / or organoleptic characteristics which have a beneficial effect on the quality of the proteins. For example, the color and viscosity of the final protein extract is beneficially affected by the method described herein. In particular, the viscosity of the purified pea protein extracts is decreased if the fermentation step described herein according to the invention is carried out, compared with protein extracts which have not been prepared according to the methods described herein. . In addition, the concentration of certain minerals in said extracts (such as potassium and magnesium, the concentration of which is decreased in the protein extracts prepared according to the processes according to the invention compared to extracts of proteins which have not not prepared according to the methods described herein) is beneficially affected by the methods described herein. It has also been found that protein extracts prepared from peas that have been fermented according to the methods of the invention described herein have a less bitter and astringent taste compared to unprepared protein extracts. according to the methods described herein.
Moreover, the presence of bacteria such as lactic acid bacteria during the hydration of peas limits the development of toxic microorganisms (due to the bacteriostatic effect of lactic acid).
Another major advantage of fermenting the peas before grinding is that after fermentation, the fermentation products, as well as the fermentation microorganisms can be easily removed and separated from the whole peas.
It has furthermore been fortuitously and fortuitously found that the characteristics of the extraction of the processes and the related process of purification are affected by the process described here. It has been found, for example, that the drop in pH, and in particular the final pH of the peas, after fermentation as defined below, reduces the undesirable overpressure in downstream equipment (as well as the associated risks of equipment damage). ). In addition, the fouling of the downstream heat exchangers which implement the downstream heat treatment is minimized, so that the cleaning frequency is decreased.
The independent and dependent claims define the particular and preferred features of the invention. The features of the dependent claims may be combined with the features of the independent claims or other dependent claims as appropriate. The appended claims are also explicitly included by reference in this specification.
BRIEF DESCRIPTION OF THE FIGURES
Figure 1 shows schematically an extraction process according to one embodiment of the invention.
Figure 2 shows a graph plotting sugar concentration / dry matter in% depending on the fermentation time of fermented peas.
Figure 3 shows a graph plotting the pH of the shelled peas and the pH of the aqueous solution (juice) as a function of the fermentation time.
FIG. 4 represents a graph plotting the acidity of the hulled peas and the acidity of the aqueous solution (juice) as a function of the fermentation time.
FIG. 5 represents a graph plotting the concentration of lactic acid bacteria of the aqueous solution (juice) contained in the first fermentation vats of a series of experiments as a function of the fermentation time.
Figure 6 is a graph plotting the viscosity profile of pea protein extracts as a function of pH.
DETAILED DESCRIPTION OF THE INVENTION
Before describing the process of the present invention, it is to be understood that this invention is not limited to the particular processes, components, products or combinations described, which methods, components, products or combinations may, of course, vary. It is also understood that the terminology used herein can not be considered restrictive, since the scope of the present invention will be limited only by the appended claims.
As used herein, the singular forms "one", "one" and "the", "the" include singular and plural references unless the context clearly indicates otherwise.
The terms "including", "includes" and "consisting of" as used herein are synonymous with "including", "included" or "containing", "contains", and are inclusive or open and do not exclude members additional elements or process steps not mentioned. It should be remembered that the terms "comprising", "includes" and "consisting of" as used herein include the terms "consists of", "consists of" and "consists of", and "substantially in "," essentially consists of "and" essentially consists of ". The evocation of numeric value ranges by their extreme points includes all the numbers and fractions integrated into the respective ranges, as well as the recited extreme points.
The term "about" or "approximately" as used herein when applied to a measurable value such as a parameter, a quantity, a time duration, and the like, means that there is a degree of variation of ± 20 % or less, preferably of ± 10% or less, more preferably of ± 5% or less, and still more preferably of ± 1% or less of the specified value, to the extent that these variations are appropriate for the realization of the present invention. It is understood that the value to which the term "about" or "approximately" refers 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, by means of a demonstration by In the example, the term includes inter alia a reference to any one of said members, or to at least two of any of said members, such as, for example, 3, 4, 5, 6 or 7 etc. of said members, and up to all said members.
All references cited herein are hereby incorporated by reference in their entirety. In particular, the teachings of all the references specifically mentioned herein are incorporated by reference. Unless otherwise defined, all terms used in the description of the invention, including technical and scientific terms, have the meaning commonly understood by any person skilled in the art of the invention. With additional information, the definitions of the terms are included to better understand the teaching of the present invention.
In the following paragraphs, different aspects of the invention are defined in more detail. Each aspect thus defined may be combined with any other aspect or aspect unless otherwise clearly indicated. In particular, any features indicated as being preferred or advantageous may be combined with one or other features indicated as preferred or advantageous.
Any reference in this description to "an embodiment" means that a particular function, structure or feature described with respect to the embodiment is included in at least one embodiment of the present invention. Thus, the occurrences of the phrase "in one embodiment" in various places of this description do not necessarily all refer to the same embodiment, but may. Moreover, the particular functions, structures or features may be combined in any appropriate manner, as would be apparent to those skilled in the art upon reading this description, in one or more embodiments. On the other hand, while some embodiments described herein include some but not other features included in other embodiments, the feature combinations of the various embodiments fall within the scope of the invention, and form different embodiments, as would be understood by 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 appended figures which form an integral part of the invention, and in which are represented by way of illustration only the specific embodiments in which the invention may be to be practiced. It is understood that other embodiments may be used and structural or logical modifications may be made without departing from the scope of the present invention. The following detailed description is therefore not to be considered as limiting, and the scope of the present invention is defined by the appended claims.
The present invention is represented in particular by any one or any combination of one or more of the following aspects and embodiments and indications numbered from 1 to 44. 1. A method of extracting pea proteins comprising the steps, preferably in the following order, comprising: (a) fermenting an aqueous composition comprising peas; (b) grinding said peas; to obtain from this fact ground peas; (c) fractionating said ground peas to obtain at least one protein fraction; and (d) isolating the pea proteins from the at least one protein fraction. 2. The method according to the indication 1, wherein said peas of step (a) are fermented until the pH of said peas is at most 5.5, preferably at most 5.0, more preferably between pH 3.5 and pH 5, measured at room temperature over 1 g of said peas which were ground and then suspended in 9 g of water. 3. The method according to claim 1 or 2, wherein said peas of step (a) are fermented until the pH of said peas is reduced by at least 1 pH unit, preferably at least 1.5 pH units, measured at room temperature over 1 g of said peas which were ground and then suspended in 9 g of water. 4. The method according to any one of claims 1 to 3, wherein step (a) comprises adding to an aqueous solution of dry peas and / or peashell, preferably dry peas having a content of dry matter between 80% and 95% based on the total weight of the dry peas. 5. The method according to any one of claims 1 to 4, wherein said peas after step (a) and before step (b) have a dry matter content of between 35% and 60% based on total weight of the peas. 6. The process according to any one of claims 1 to 5, wherein said peas of step (a) are fermented for at least 3 hours, preferably for at least 3 hours and not more than 24 hours. 7. The process according to any one of the indications 1 to 6, wherein said peas of step (a) are subjected to fermentation at a temperature of between 30 ° C. and 50 ° C., preferably between 35 ° C. and 45 ° C. 8. The process according to any one of claims 1 to 7, wherein step (a) comprises the fermentation of said peas in the presence of lactic acid bacteria, preferably in the presence of one or more species of lactobacilli. 9. The method according to any one of the indications 1 to 8, wherein said peas of step (a) are subjected to fermentation in the presence of at least 102 cfu at 1010 cfu of lactic acid bacteria per ml of said composition. aqueous composition comprising peas. The process according to any one of claims 1 to 9, wherein the fractionation of said ground peas in step (c) comprises adjusting the pH of the ground peas to a pH of at least 6, preferably from at least 7, preferably at a pH of at least 8 and at most 9. This pH adjustment can be carried out using any suitable base, such as sodium hydroxide, potassium hydroxide, calcium hydroxide. Preferably, this pH adjustment is carried out on an aqueous composition comprising ground peas having a dry matter content of at most 45%, preferably at most 40%, preferably at most 35%, preferably at most 30%, preferably at most 25%. In one embodiment, the dry matter content of the ground peas is adjusted to the above dry matter content by adding water accordingly. 11. The method according to any one of claims 1 to 10, wherein the fractionation of said ground peas in step (c) comprises subjecting said ground peas to one or more separation steps, preferably one or more decantation steps. preferably one or more centrifugation decantation steps. The process according to any of claims 1 to 11, wherein isolating the pea protein from said protein fraction in step (d), comprises at least one step selected from the following: precipitation, flocculation, filtration, and / or chromatography. 13. The pea proteins obtainable by the method according to any one of the 1 to 12 indications. 14. An edible composition, preferably a product intended for human or animal consumption, comprising pea proteins. according to the indication 13. 15. The use of pea protein according to the indication 13 in products intended for human or animal consumption, preferably in dairy products, confectionery products, beverages, products meat products, vegetarian products, dietary supplements, nutritional and weight-control products, foods for medical purposes, foods for the elderly and bakery products. 16. The process of any one of claims 1 to 12 wherein step (a) comprises contacting the shelled peas with an aqueous solution. 17. The process according to any one of claims 1 to 12, 16, wherein step (a) comprises contacting the dry shelled peas with an aqueous solution, preferably dry shelled peas having a content content between 80% and 95% based on the total weight of dry husked peas. 18. The method according to any of the indications 16 or 17, wherein said peas after step (a) and before step (b) have a dry matter content of between 40% and 50% based on total weight of the peas. 19. The process according to any one of claims 1 to 12, 16 to 18, wherein before, during and / or after the grinding step (b) an aqueous solution is added, preferably water, preferably to obtain an aqueous composition comprising ground peas, said composition comprising from 15% to 35% of dry matter on the basis of the total weight of the composition, preferably from 15% to 35%, preferably from 18% to 33%, for example from 20% to 30%, such as at least 20%, for example at least 21%, for example at least 22%, for example at least 23%, for example at least 24%, for example at least 25%, 26%, 27%, 28%, 29%, for example at most 30%, for example at most 35%. 20. The method according to any of the indications 1 to 12, 16 to 19, wherein said peas of step (a) are subjected to fermentation for not more than 24 hours, for example not more than 20 hours, by for not more than 18 hours, for example for not more than 12 hours, for example not more than 10 hours. 21. The method according to any one of the indications 1 to 12, 16 to 20, wherein at the end of step (a) said peas have an acidity of between 25 and 250 mEq OH 'per gram of pea. 22. The process according to any one of Claims 1 to 12, 16 to 21, wherein at the end of step (a) said peas have a sugar content of not more than 6.0% by weight on the basis of the total dry matter content of said peas, the sugar content being the total amount of glucose, fructose, sucrose, verbascose, raffinose, stachyose, and galactose; preferably at most 5.5%, for example at most 5.0%, for example at most 4.5%, for example at most 4.0%. 23. The method according to any one of claims 19 to 22, wherein the fractionation of said ground peas to obtain at least one protein fraction in step (c) comprises adjusting the pH of the aqueous composition comprising pea milled to a pH of at least 6, preferably at least 7, preferably at least 8, preferably at a pH of at least 7.5 and at most 9, preferably at least a pH of at least 7.5 and at most 8.5. Preferably, this pH adjustment is carried out on an aqueous composition comprising ground peas having a dry matter content of at most 45%, preferably at most 40%, preferably at most 35%, preferably at most 30%, preferably at most 25%. In one embodiment, the dry matter content of the ground peas is adjusted to the above dry matter content by adding water accordingly. 24. The process according to any one of claims 1 to 12, 16 to 23, wherein said at least one protein fraction is subjected to a temperature of at least 30 ° C, for example at least 40 ° C, for example at least 50 ° C, for example at least 55 ° C, for example at most 80 ° C, for example at least 50 ° C and at most 80 ° C, for example at least 53 ° C and at most 78 ° C, for example at least 54 ° C and at most 75 ° C. 25. The process according to any one of claims 1 to 12, 16 to 24, wherein said aqueous composition comprising peas in step (a), comprises an aqueous solution, preferably water. 26. The method according to any one of the indications 1 to 12, 16 to 25, wherein the amount of pea in said aqueous composition comprising peas is preferably between 150 and 500 kg of peas per m 3 of aqueous composition comprising peas. 27. The process according to any one of claims 1 to 12, 16 to 26, wherein said aqueous composition comprising peas before or at the beginning of the fermentation step (a) has a pH of at least 6, by example of at least 6.2, for example at least 6.4, measured on the aqueous composition comprising peas, after said composition has been milled. 28. The method according to any one of the indications 1 to 12, 16 to 27, wherein said lactic acid bacteria are chosen from the group comprising: Lactobacillus, Leuconostoc, Pediococcus, Streptococcus, Aerococcus, Carnobacterium, Enterococcus, Oenococcus, Sporolactobacillus, Tetragenococcus , Vagococcus, and Weisella, and their combinations. 29. The method according to any one of claims 1 to 12, 16 to 28, wherein the lactic acid bacteria are lactobacillus species, preferably selected from the group consisting of: Lactobacillus fermentum, Lactobacillus crispatus, Lactobacillus panis, Lactobacillus mucosae , Lactobacillus pontis, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus helveticus, Lactobacillus buchneri, Lactobacillus delbrueckii and Lactobacillus casei and mixtures thereof. 30. The process according to any one of claims 1 to 12, 16 to 29, wherein the lactic acid bacteria are selected from the group consisting of: Lactobacillus fermentum, Lactobacillus crispatus, Lactobacillus panis, Lactobacillus mucosae, Lactobacillus pontis, and mixtures thereof. 31. The process according to any one of claims 1 to 12, 16 to 30, wherein the lactic acid bacteria are selected from the group consisting of: Lactobacillus fermentum, Lactobacillus crispatus, Lactobacillus panis, Lactobacillus mucosae, Lactobacillus pontis, and mixtures thereof. 32. The method of any one of claims 1 to 12, 16 to 31, wherein said lactic acid bacteria are Lactobacillus fermentum, or Lactobacillus crispatus. 33. The process of any one of 1 to 12, 16 to 32, wherein said fermentation is anaerobic fermentation. 34. The method according to any one of claims 1 to 12, 16 to 33, wherein the dry peas before the start of step (a) have a pH of at least 6.0, preferably an pH inclusive of between 6.0 and 7.0, such as, for example, a pH of at least 6.0, for example at least 6.1, for example at least 6.2, for example at least 6, 3, for example at most 6.9, for example at most 7.0, preferably between 6.25 and 6.75, measured at room temperature over 5 g of dry peas which have been milled with 95 g of water. 35. The process of any one of 1 to 12, 16 to 34, comprising the steps of: (a) fermenting an aqueous composition comprising peas; (b) grinding said peas in the presence of water; to thereby obtain an aqueous composition comprising ground peas; (c) fractionating said aqueous composition comprising ground peas to obtain at least one protein fraction, preferably by adjusting the pH of said aqueous composition to a pH of at least 6; preferably, this pH adjustment is carried out on the aqueous composition comprising ground peas having a dry matter content of at most 45%, preferably at most 40%, preferably at most 35%, preferably at most 30%, preferably at most 25%. In one embodiment, the dry matter content of the aqueous composition is adjusted to the above-mentioned dry matter content by adding water accordingly; (d) isolating the pea proteins from the at least one protein fraction. 36. The process according to any one of claims 1 to 12, 16 to 35, wherein step (c) comprises fractionating said ground peas into a fraction comprising at least 50% by weight of proteins on the basis of total dry matter content of said fraction. 37. The method according to any one of claims 1 to 12, 16 to 36, wherein isolating the pea protein from said protein fraction in step (d) comprises a single precipitation step. 38. The method according to any one of Claims 1 to 12, 16 to 37, wherein the isolation of the pea proteins from said protein fraction in step (d) is carried out by isoelectric precipitation. 39. The process according to any one of Claims 1 to 12, 16 to 38, wherein step (d) further comprises: (e) obtaining said isolated pea proteins as an aqueous suspension; (f) optionally subjecting said aqueous suspension to at least one heat treatment; (g) optionally drying said aqueous suspension. 40. The method according to the indication 39, wherein step (e) further comprises adjusting the pH of said aqueous suspension to a pH of at least 6.0, preferably the pH is adjusted to a pH at least 6.5, preferably between pH 6.0 and 8.5, preferably between pH 6.5 and 8.5, preferably between pH 7.0 and 8.5. 41. The process according to any one of the indications 39 or 40, wherein step (f) comprises: subjecting said aqueous suspension to a heat treatment for at least 0.01 s, preferably for a duration of between 0, 01 s and 20 min, preferably between 10 s and 10 min. 42. The process according to any one of the indications 39 to 41, wherein said heat treatment in step (f) is carried out at a temperature of at least 70 ° C, preferably at a temperature between 75 ° C and 210 ° C, preferably between 85 ° C and 160 ° C, for example between 90 ° C and 150 ° C. 43. The process according to any one of Claims 1 to 12, 16 to 42, wherein before step (a) said dry peas have a sugar content of at least 6.2% by weight based on the total dry matter content of said peas, the sugar content being the total amount of glucose, fructose, sucrose, verbascose, raffinose, stachyose, and galactose. 44. The process according to any one of Claims 1 to 12, 16 to 43, wherein step (d) comprises isolating said pea protein in the form of an extract comprising at least 60% by weight of preferably at least 70% by weight, more preferably at least 80% by weight, for example at least 85% by weight of protein on the basis of the total dry matter content of said extract.
In a first aspect, the invention relates to a method of extracting pea proteins, comprising the steps of: (a) fermenting an aqueous composition comprising peas; (b) grinding said peas; (c) fractionating said ground peas to obtain at least one protein fraction; (d) isolating the pea proteins from the at least one protein fraction.
As used herein, the term "pea" refers to the round seeds contained in the pod of Pisum sativum and its subspecies, varieties or cultivars. Preferably, the peas are yellow peas, preferably dry yellow peas, i.e. yellow peas that have been harvested in the dry state. The term "pea protein" as used herein, therefore, refers to the proteins contained in pea seeds.
According to the invention, the peas may be whole peas, that is to say peas as they are present in the pod. In a preferred embodiment, however, the peas are shelled peas, i.e. peas whose integument has been removed. Peas are peeled peas where the outer layer of the seed is removed. Removal of the integument can be achieved by techniques known in the art, such as mechanically with the aid of hullers. It is understood that when reference is made here to shelled peas, in some embodiments, not all but nevertheless the vast majority of the individual peas are shelled, preferably more than 90% peas.
Peas, as used herein, can be sorted before being fermented. For example pebbles or larger plant material, but also damaged peas, can be removed from peas for use according to the invention.
As used herein, the term "extracting pea proteins" refers to releasing and separating pea proteins from other pea constituents. Pea protein extraction according to some embodiments of the present invention. The invention may include isolation or purification of pea proteins. Those skilled in the art will appreciate that pea protein extracts are not entirely protein-based, and that a certain amount of additional components (impurities) may be present in pea protein extracts, such as lipids, carbohydrates , sugars, minerals, etc.
As used herein, the term "sugar" or "free sugar" refers to mono-, di-, and / or oligosaccharides consisting of not more than 10 monomer units. In certain embodiments, the terms "total sugars" or "total free sugars" refer to all mono-, di-, and / or oligosaccharides consisting of not more than 10 monomer units. In other embodiments, a specific subset of sugars is indicated.
In some embodiments of the invention, the pea protein extracts comprise, on a dry matter basis, at least 50% by weight protein (ie 50 g protein for a total of 100 g of dry matter), preferably at least 75% by weight of proteins. In some embodiments, the pea protein extracts comprise, on a dry matter basis, at least 50% by weight to at most 95% by weight or 99% by weight of proteins, such as at least 75% by weight. at most 99% by weight of protein. The crude extracts generally comprise a lower fraction of proteins than the refined or purified extracts.
According to the invention, the steps (a) to (d) of the method specified above are preferably carried out in the following order: step (a) precedes step (b), which itself precedes step (c), which itself precedes step (d). However, it is also possible according to the invention to perform steps (b) and (c) simultaneously, that is to say that the grinding step and the fractioning step are performed simultaneously.
In step (a) of the process described herein, an aqueous composition comprising peas is fermented. According to the invention, the peas which are fermented in step (a) are unmilled peas (i.e., whole peas). Peas may, however, in one embodiment be broken peas. In one embodiment, the peas are round at harvest and after drying. After removing the integument, the natural pieces of seed cotyledons can be separated manually or mechanically, to give "split peas".
As used herein, the term "aqueous composition comprising peas" used in step (a) refers to a composition consisting mainly or exclusively of an aqueous solution such as water, in addition to peas. . In some embodiments, the aqueous composition includes, for example, a pea suspension in an aqueous solution. In a preferred embodiment, the aqueous solution is water. In one embodiment, the water may be tap water, or well water that has been treated to make it potable. The water used is preferably drinking water, that is to say water suitable for human consumption.
In some embodiments, the amount of pea that is added to the aqueous solution to reconstitute the aqueous composition comprising peas is preferably between 150 and 500 kg peas per m3 of aqueous composition comprising peas, that is, for 150 to 500 kg of peas an aqueous solution is added until a final volume of 1 m 3 is obtained.
In one embodiment, the aqueous composition comprising peas at the beginning of step (a) of the above described here, preferably at the start of fermentation, has a pH of at least 6, preferably at least 6. , 2, for example of at least 6.4, measured on the aqueous composition comprising peas, after said composition has been milled.
In a preferred embodiment, the peas which are in contact with the aqueous composition are harvested naturally dry, or in one embodiment the peas may be fresh peas. Preferably, the peas are dry peas, and have a solids content (by weight) of at least 80% (ie at least 80 g of dry matter for a total weight of 100 g of dry peas), more preferably at least 85%, for example at least 90%, for example at least 95%, for example a solids content of between 80% and 95%, for example between 85% and 95%, for example between 90% and 95%.
As used herein, the term "fermentation" has the meaning commonly accepted in the art. Additional information states that fermentation is a microbiological metabolic process involving the conversion of sugars to acids, and / or gas using yeasts and / or bacteria. Subjecting an aqueous composition comprising peas to fermentation, as used herein, can therefore refer to incubating the aqueous composition comprising peas with bacteria and / or yeasts under appropriate conditions for the bacteria and / or the yeasts are metabolically active.
In one embodiment, the aqueous composition comprising peas is fermented in step (a) of the method described above with lactic acid bacteria. As used herein, the term "lactic acid bacteria" refers to a population of cocci or gram-positive, low-guanine + cytosine, acid-tolerant, generally non-sporulated, anaerobic bacilli that are associated by their metabolic characteristics. and physiological and produce lactic acid as the main metabolic end product of carbohydrate fermentation. These bacteria can usually be present in decaying plants and milk products. As used herein, lactic acid bacteria can be non-pathogenic in that they do not cause damage or lead to deleterious effects if ingested. Preferably, the lactic acid bacteria, as used herein, are one or more bacterial genera selected from Lactobacillus,
Pediococcus, Lactococcus, Leuconostoc, Streptococcus, Aerococcus, Carnobacterium, Enterococcus, Oenococcus, Sporolactobacillus, Tetragenococcus, Vagococcus, and Weisella, and combinations thereof. Most preferably, the lactic acid bacteria are lactobacillus species, most preferably selected from the group consisting of: Lactobacillus fermentum, Lactobacillus crispatus, Lactobacillus parties, Lactobacillus mucosae, Lactobacillus pontis, Lactobacillus acidophilus, Lactobacillus planta rum, Lactobacillus Helveticus, Lactobacillus buchneri, Lactobacillus delbrueckii, and Lactobacillus casei, and mixtures thereof, for example in the group consisting of: Lactobacillus fermentum, Lactobacillus crispatus, Lactobacillus panis, Lactobacillus mucosae, Lactobacillus pontis, Lactobacillus acidophilus and mixtures thereof, for example in the group consisting of: Lactobacillus fermentum, Lactobacillus crispatus, Lactobacillus panis, Lactobacillus mucosae, Lactobacillus pontis, and mixtures thereof, for example said bacteria are Lactobacillus fermentum, or Lactobacillus crispatus. In some embodiments, the fermentation may be spontaneous fermentation (i.e., wherein no fermentation microorganism is deliberately added, but the fermentation is carried out by microorganisms that are naturally present on / in peas and / or in the environment) or inoculated fermentation (that is to say in which fermentation microorganisms are deliberately added). The fermentation can also be carried out by transferring part or all of the aqueous fraction of a fermentation step to a subsequent fermentation which will be started later, for example by transferring at least 1 / 10th of the volume of the first fermentation to at least a second fermentation step. In a preferred embodiment, the fermentation is anaerobic fermentation.
In one embodiment, the aqueous composition comprising peas is fermented in step (a) of the process described above until the pH of the peas is at most 5.5, preferably of at most 5.0, more preferably between 3.5 and 5, preferably measured at room temperature over 1 g of said peas which have been ground and then suspended in 9 g of water, as described in FIG. Experimental part. In one embodiment, the aqueous composition comprising peas is fermented in step (a) of the method described above until the pH of the peas is between 3.5 and 4.5, for example between 4.0 and 5.0, preferably between 4.5 and 5.5, such as for example a pH of at least 3.5, for example at least 3.75, for example from minus 4.0, for example at least 4.25, for example at least 4.50, for example at least 4.75, for example at most 5.0, for example at plus 5.25, for example at most 5.5, preferably measured at room temperature over 1 g of said peas which have been ground and then suspended in 9 g of water, as described in the experimental part.
In one embodiment, the dry peas have a pH of at least 6.0, preferably between 6.0 and 7.10, before being fermented in step (a) of the method described above, as for example a pH of at least 6.0, for example at least 6.1, for example at least 6.2, for example at least 6.3, for example 6.4, for example example 6.5, for example 6.6, for example 6.7, for example 6.8, for example 6.9, for example 7.10, preferably between 6.25 and 6.75, preferably measured at room temperature over 5 g of dry peas which were ground with 95 g of water.
In one embodiment, the aqueous composition comprising peas is fermented in step (a) of the process described above until the pH of the peas decreases by at least 1 pH unit, preferably at least 1.5 pH units, such as for example at least 1, for example at least 1.1, for example at least 1.2, for example at least 1.3. for example at least 1.4, for example at least 1.5, for example at least 1.6, for example at least 1.7, for example at least 1.8 for example at least 1.9, for example at least 2, for example at least 2.1, for example at least 2.2, for example at least 2.3, for example example of at least 2.4, for example at least 2.5, for example at least 2.6, for example at least 2.7, for example at least 2.8, for example example of at least 2.9, for example at least 3 pH units, preferably measured at room temperature over 1 g of said peas which have been ground and then suspended in 9 g of water. In another embodiment, the aqueous composition comprising peas is fermented in step (a) of the method described above until the pH of the peas decreases from 1 pH unit to 3 pH, preferably 1.5 pH units at 3 pH units, for example 1.5 pH units at 2.5 pH units, for example 2.0 pH units at 3.0 units of pH pH, preferably measured at room temperature over 1 g of said peas which have been ground and then suspended in 9 g of water. By way of example, and without limitation, at the start of the fermentation, the pH of the peas may be 6.5, and at the end of the fermentation, the pH of the peas may be 5.0, preferably measured. at room temperature over 1 g of said peas which were ground and then suspended in 9 g of water, as described in the experimental part.
In one embodiment, the aqueous composition comprising peas is subjected to fermentation in step (a) of the method described above for a period of at least 3 hours, preferably at least 4 hours, plus preferably at least 6 h. In another embodiment, the aqueous composition comprising peas is subjected to fermentation in step (a) of the process described above for a period of between 3 hours and 24 hours, preferably between 4 hours and 24 hours. more preferably between 4 h and 20 h, such as for example a duration of at least 3 h, for example at least 4 h, for example at least 5 h, for example at least 6 h, by at least 7 hours, for example at least 8 hours, at least 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, for example not more than 15 hours, for example not more than 16 hours, for example not more than 17 hours, for example not more than 18 hours, for example not more than 19 hours, by example of not more than 20 hours, for example not more than 21 hours, for example not more than 22 hours, for example not more than 23 hours, for example not more than 24 hours. Those skilled in the art will appreciate, for example, that spontaneous fermentation may take longer than fermentations which are carried out by adding bacteria, depending on the different amounts of microorganisms at the start of fermentation.
In one embodiment, the pea-containing aqueous composition is subjected to fermentation in step (a) of the method described above at a temperature that is optimal for the fermentation microorganisms, preferably at a temperature which is at most 5 ° C above or below the optimum temperature for fermentation microorganisms. Optimum temperatures for bacteria and / or yeasts as defined herein are known in the art. Additional information specifies, without limitation, that an optimum temperature, as defined herein, refers to the temperature at which growth is maximal. In a further embodiment, the aqueous composition comprising peas is subjected to fermentation in step (a) of the process described above at a temperature of at least 30 ° C, for example between 30 ° C and 50 ° C, preferably between 35 ° C and 45 ° C. In another embodiment, the aqueous composition comprising peas is fermented in step (a) of the process described above at a temperature of between 30 ° C and 40 ° C, between 35 ° C and 45 ° C. ° C, or between 40 ° C and 50 ° C, preferably a temperature of 40 ° C or about 40 ° C.
In one embodiment, the aqueous composition comprising peas is subjected to fermentation in step (a) of the method described above in the presence of fermentation microorganisms, such as bacteria and / or yeasts, preferably comprising one or more lactic acid bacteria and / or yeasts, more preferably said fermentation microorganisms are selected from the group consisting of one or more species of lactobacilli and / or yeasts. In one embodiment, the fermentation is carried out in the presence of one or more of the microorganisms specified above at a concentration of between 102 cfu / ml and 1010 cfu / ml of said aqueous composition comprising peas, such as a concentration of at least 102 ug / ml, for example at least 105 ug / ml, for example at least 106 ug / ml, for example at least 107 ug / ml, for example at least 108 ug / ml, for example from at least 109 cfu / ml of said aqueous composition comprising peas. "Cfu" units (colony forming unit) are well known in the art and can be determined for example by plate counting. It is understood that "cfu / ml" refers to the amount of colony forming units per ml of the aqueous composition comprising total peas, i.e. including peas.
In another embodiment, the aqueous composition comprising peas is subjected to fermentation in step (a) of the method described above in the presence of fermentation microorganisms, preferably comprising one or more lactic acid bacteria and / or or yeasts, more preferably comprising one or more species of lactobacilli, wherein the microorganisms are added at a concentration of at least 102 cfu / ml of the aqueous composition comprising peas.
In one embodiment, the peas after step (a) and before step (b) of the method described above, that is to say at the end of the fermentation and before the grinding step have a dry matter content (by weight) of between 35% and 60%, preferably between 35% and 55%, for example between 40% and 50%, for example a solids content of at least 40% %, for example at least 41%, at least 42%, for example at least 43%, for example at least 44%, for example at least 45%, for example from at least 46%, for example at least 47%, about 48%, about 49%, for example not more than 50%, for example not more than 55%, for example not more than 60% % on the basis of the total weight of the peas at the end of the fermentation, i.e. after the peas have been isolated from the aqueous composition.
In step (b) of the method according to the invention described above, the peas which have been subjected to fermentation in step (a) are milled. For this purpose, in one embodiment, the peas are removed from the aqueous composition after step (a) and then subjected to milling. Preferably, the peas are washed or rinsed after step (a) and before step (b). The washing and rinsing can be carried out with an aqueous solution, preferably water, such as tap water, or treated well water, preferably drinking water, that is, to say clean water for human consumption.
As used herein, the term "grinding" has the meaning commonly accepted in the art. Additional information specifies that grinding, as used herein, may refer to the process of milling solids i.e. peas, by exposure to mechanical forces that destroy the structure by overcoming bonding forces internal. The grinding can thus disintegrate the original structure of the peas. In a preferred embodiment, the size of the ground pea particles comprising at least 25% of dry matter has a D50 of at most 300 μm, preferably at most 250 μm, for example at most 200 μm, the D50 being defined as the particle size for which fifty percent by volume of the particles are smaller than D50; and the D50 being measured by laser diffraction analysis on a Malvern analyzer.
For example, the D50 can be measured by sieving or laser diffraction analysis. For example, Malvern Instruments laser diffraction systems can be advantageously used. Particle size can be measured by laser diffraction analysis on a Malvern analyzer. Particle size can be measured by laser diffraction analysis on a Malvern type analyzer after the peas have been milled and incorporated into an aqueous slurry having a solids content of 25%. Suitable Malvern systems include Malvern 2000 instruments, Malvern MasterSizer 2000 (such as Mastersizer S), Malvern 2600 and Malvern 3600. These instruments and their user manuals meet or exceed the requirements of ISO 13320. Malvern MasterSizer instrument (such as Mastersizer S) can also be useful because it can more accurately measure the D50 towards the lower end of the range eg for average particle sizes of less than 8 pm, applying the theory of Mie, using appropriate optical means.
In one embodiment, before, during, or after grinding the peas in step (b) of the process according to the invention described above, an aqueous solution, preferably water, such as tap water, or treated well water, preferably drinking water, i.e., water suitable for human consumption, is added to the peas. In a further embodiment, an amount of aqueous solution is added to the peas so as to obtain an aqueous composition comprising ground peas, said composition preferably comprising between 15% and 35% of dry matter on the basis of the total weight of the composition, preferably between 15% and 35%, preferably between 20% and 30%, as at least 19%, as at least 20%, as at least 21%, as at least 22%, for example at least 23% for example at least 24%, for example at least 25%, for example at least 26%, for example at least 27%, for example at least 28%, for example at least 29%, for example at most 30%, for example at most 35% dry matter on the basis of the total weight of the composition. In a preferred embodiment, the milling process is a wet grinding process, such that an aqueous solution is added to the peas before or during milling.
In one embodiment, step (c) of the method according to the invention described above comprises fractionating said ground peas into a fraction comprising at least 50% by weight of proteins on the basis of the total content of the material. of said fraction. As used herein, the term "fractionation" refers to a process by which at least a portion of the proteins included in the peas are separated from the rest of the peas. It is understood that when reference is made to the fractionation step, in some embodiments, not all but nevertheless the vast majority of the individual proteins are separated, preferably at least 50% by weight, preferably at least 60% by weight. % by weight of the proteins, on the basis of the total protein content of the ground peas are separated.
Fractionation of ground peas into a protein fraction can be accomplished by any means known in the art, such as the addition of a suitable base or salt.
Preferably, the ground peas are fractionated by increasing the pH of the ground peas. Preferably, the fractionation step (c) comprises adjusting the pH of the ground peas to a pH of at least 6, preferably at least 7, most preferably at a pH of at least 8. and at most 9. Preferably, the fractionation step (c) comprises increasing the pH of an aqueous composition comprising ground peas. In a preferred embodiment, the pH of the composition is adjusted to a pH of at least 6, more preferably at least 7. In another preferred embodiment, the pH of the composition is adjusted to a value of between pH 6 and pH 9, more preferably between pH 7 and pH 9, as a pH of at least 7.0, for example at least 7.1, for example at least 7.2, for example at least 7.3, for example at least 7.4, for example at least 7.5, for example at least 7.6, for example at least 7.7, e.g. at least 7.8, for example at least 7.9, for example at least 8.0, for example at least 8.1, for example at least 8.2, e.g. at least 8.3, for example at least 8.4, for example at most 8.5, for example at most 8.6, for example at most 8.7, e.g. at most 8.8, for example at most 8.9, for example at most 9.0, most preferably at pH between pH 7.5 and pH 8.5, preferably between t pH 8 is preferably applied to an aqueous composition comprising ground peas having a dry matter content of at most 45%, preferably at most 40%, preferably at least 40% by weight. at most 35%, preferably at most 30%, preferably at most 25%. In one embodiment, the dry matter content of the ground peas is adjusted to the above dry matter content by adding water accordingly. This pH adjustment can be carried out using any suitable base, such as sodium hydroxide, calcium hydroxide, potassium hydroxide and the like. In a preferred embodiment, the pH of the ground pea-containing compositions is adjusted by adding sodium hydroxide.
In a preferred embodiment, after adjusting the pH, the protein fraction is separated from the aqueous composition comprising ground peas, by decantation or by using a hydrocyclone, preferably by decantation, preferably by centrifugal decantation (that is, ie by means of a decanting centrifuge), the protein fraction being the supernatant, and the centrifugation pellet being a fraction comprising inter alia the rest of the ground peas and residual proteins. In one embodiment, several fractionation steps can be performed successively. For example, after decantation, the centrifugation pellet may be suspended in an aqueous solution (preferably in an aqueous solution preferably having a similar or higher pH (preferably a pH of 8.5 or about 8.5 ) to that of the first fractionation step) and subjected to a decantation step, so as to recover additional proteins in the supernatant.
As indicated elsewhere in this document, step (c) and step (b) of the process according to the invention can be carried out simultaneously or alternatively, step (c) can be carried out as a result of step (b).
It is understood that the protein fraction may also comprise other constituents, in particular those which are rendered soluble or remain soluble during the fractionation step.
In one embodiment, the protein fraction comprises at least 1.0% dry matter based on the total weight of the composition, preferably at least 2.0% dry matter, more preferably at least 3.0% by weight. dry matter, such as for example at least 4.0% dry matter, such as for example at least 5.0% dry matter.
In another embodiment, the protein fraction comprises from 1.0% to 40% of dry matter, preferably from 2.0% to 30% of dry matter, more preferably from 3.0% to 20% of dry matter. more preferably from 3.0% to 15% of dry matter, such as from 3.0% to 10%.
In one embodiment, the dry matter of the protein fraction comprises at least 50% by weight of pea protein, preferably at least 60% by weight of pea protein, more preferably at least 65% by weight of pea protein. as for example at least 70% by weight, such as at least 55% by weight and at most 80% by weight, or between 60% by weight and 80% by weight, or between 60% by weight and 78% by weight.
In some embodiments, in a further step, the protein fraction is subjected to at least one heat treatment, preferably said protein fraction is subjected to a temperature of at least 30 ° C, for example at least 40 ° C for example at least 50 ° C, for example said protein fraction is subjected to a temperature between 30 ° C and 90 ° C, more preferably between 50 ° C and 80 ° C, still more preferably between 55 ° C and 75 ° C, such as, for example, 55 ° C, 60 ° C, 65 ° C, 70 ° C, or 75 ° C. In one embodiment, the temperature of the heat treatment is between 50 ° C and 60 ° C, for example between 55 ° C and 65 ° C, for example between 60 ° C and 70 ° C, for example between 65 ° C and 75 ° C, or for example between 70 ° C and 80 ° C.
In step (d) of the method according to the invention described above, the pea proteins are isolated from said protein fraction. As used herein, the term "isolated" or "isolate" may refer to a process that separates proteins from said protein fraction. The terms "concentration" and "isolation" can be used interchangeably. Preferably, said isolation step can be carried out by precipitation, flocculation, filtration, and / or chromatography, or by a combination thereof.
In one embodiment, step (d) of the method of the invention described above, includes additional post-treatment steps to further purify the pea proteins and / or increase the yield. Basically speaking, these additional steps can be used to remove impurities that are coarse with isolated proteins.
One or more of the following steps, preferably all, may be implemented for this purpose.
In some embodiments, the proteins are isolated from said aqueous composition comprising pea proteins by precipitation, flocculation, filtration, and / or chromatography. Preferably, the proteins are isolated by isoelectric precipitation or ultrafiltration. In a preferred embodiment, the isolation of pea proteins from said protein fraction in step (d) comprises at least one isoelectric precipitation step of said proteins. In a preferred embodiment, isolating pea proteins from said protein fraction in step (d) comprises a single step of isoelectric precipitation of said proteins.
Preferably, the pH of the protein fraction is adjusted to the isoelectric point of the proteins. As used herein, the term "isoelectric point" refers to the pH at which the proteins have a net ionic charge of 0, or substantially 0 (i.e. the sum of the negative and positive charges is equal to 0). , or substantially 0). It is understandable that the isoelectric point of the individual proteins can vary. As used herein, the isoelectric pH of the protein fraction as used herein refers to the pH of the fraction for which the overall protein load in the fraction is 0, or substantially 0. The isoelectric pH of the proteins and Protein compositions can be determined by techniques known in the art. The isoelectric pH is here determined as the pH at which the solubility index of nitrogen is the lowest. In a preferred embodiment, the pH of the protein fraction is adjusted in a range from 4.0 to 5.8, preferably from 4.5 to 5.5, preferably from 4.5 to 5.0, for example at a pH of 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8. PH adjustment can be performed by adding an acid, such as sulfuric acid, or hydrochloric acid. At the isoelectric point, most proteins precipitate or form aggregates.
Separation of the precipitated or aggregated proteins can be carried out by decantation, preferably by decantation by centrifugation. In a preferred embodiment, the dry matter content (by weight) after separation of the precipitated or aggregated proteins is between 20% and 40%, for example it is at least 25%, for example at least 26%. %, for example at least 27%, for example at least 28%, for example at least 29%, 30%, 31%, 32%, 33%, 34%, or 35%, preferably at least 27% and not more than 38% on the basis of the total weight of the precipitated or aggregated proteins (also referred to as "aqueous suspension"). The dry matter content can be further adjusted, for example by adding an aqueous solution to the precipitated or aggregated proteins, thereby obtaining a precipitated protein composition. In one embodiment, said aqueous solution is preferably water, preferably drinking water, that is to say water suitable for human consumption. Preferably, the dry matter content can be adjusted to a range of between 10% and 25%, preferably between 15% and 20%, it can be for example at least 15%, for example at least 16%. %, preferably at least 17%, 18%, 19%, 20% based on the total weight of the precipitated protein composition. Optionally, the protein concentration step may be repeated at least one more time. Preferably, the protein concentration step is performed once.
In a preferred embodiment, the precipitated or aggregated proteins are preferably resuspended in an aqueous solution, preferably water, preferably potable water, that is clean water. for human consumption. The dry matter content is preferably between 10% and 25%, preferably between 15% and 20%, it is for example at least 15%, for example at least 16%, 17%, 18% , 19%, 20% of the resuspended protein composition.
In one embodiment, the pH of the resuspended protein composition is adjusted to a pH of at least 6.0, preferably the pH is adjusted to a pH of at least 6.5, preferably between pH 6.0 and 8.5, preferably between pH 6.5 and 8.5, preferably between pH 7.0 and 8.5, preferably between pH 7.3 and 8.0, as for example at a pH of at least 7.2, for example at least 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0. For this purpose, sodium hydroxide for example, or any suitable base may be used to adjust the pH to the desired level. Preferably, this pH adjustment is performed on an aqueous composition comprising resuspended proteins having a dry matter content of at most 45%, preferably at most 40%, preferably at most 35%, preferably at most 30%, preferably at most 25%. In one embodiment, the dry matter content of the aqueous composition comprising resuspended proteins is adjusted to the above-mentioned dry matter content by adding water accordingly.
In another embodiment, the pH of the resuspended protein composition is adjusted to a range of pH 4.0 to 5.8, preferably pH 4.5 to 5.5, such as pH 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5. For this purpose, sodium hydroxide or sulfuric acid, for example, may be used to adjust the pH to the desired level. Preferably, this pH adjustment is performed on an aqueous composition comprising resuspended proteins having a dry matter content of at most 45%, preferably at most 40%, preferably at most 35%, preferably at most 30%, preferably at most 25%. In one embodiment, the dry matter content of the aqueous composition comprising resuspended proteins is adjusted to the above-mentioned dry matter content by adding water accordingly.
Optionally, but preferably, the resuspended protein composition, whose pH has been set in the range indicated above, is further subjected to at least one heat treatment, preferably a heat treatment at a temperature of at least 70 ° C, preferably at least 75 ° C, more preferably at least 80 ° C, still more preferably at least 85 ° C, even more preferably at least 90 ° C, for example at least 95 ° C, preferably at most 160 ° C, still more preferably at most 210 ° C. For example, the temperature of said heat treatment may be between 70 ° C and 210 ° C, preferably between 85 ° C and 160 ° C, more preferably between 90 ° C and 150 ° C. The heat treatment can be advantageously carried out by means of one or more heat exchangers or by direct or indirect injection of steam. In one embodiment, the duration of the heat treatment is at least 0.01 s, preferably between 0.01 s and 20 min, preferably between 10 s and 10 min. Those skilled in the art will appreciate that the higher the temperature, the shorter the duration of the heat treatment. For example, the temperature of the heat treatment may be between 115 ° C and 210 ° C for a time between 0.01 s and 15 s. Alternatively, for example, the heat treatment can be carried out at a temperature between 95 ° C and 115 ° C for a period of between 15 s and 5 min. Alternatively, for example, the temperature of the heat treatment may be between 70 ° C and 95 ° C for a time between 5 min and 15 min. In a preferred embodiment, the heat treatment is carried out at a temperature between 95 ° C and 110 ° C for a time between 2 min and 8 min. In another preferred embodiment, the heat treatment is carried out at a temperature between 130 ° C and 140 ° C for a period of between 1s and 8s. After the heat treatment, the compositions comprising proteins can be maintained at a temperature between 70 ° C and 90 ° C, preferably between 70 ° C and 85 ° C, before drying.
In yet another additional step, the compositions comprising proteins may be subjected to drying, whether or not they have been previously subjected to heat treatment after isolation, or whether or not they have been previously subjected to precipitation. Drying can be carried out by any means known in the art, such as by hot air application, evaporation, lyophilization, contact drying, steam drying, dielectric drying, cylinder drying, flash drying, etc. In a preferred embodiment, the proteins are dried by spray drying. Optionally, compositions comprising proteins may be granulated by techniques known in the art.
In a preferred embodiment, the present invention relates to a method of extracting pea proteins of the species Pisum sativum, comprising the steps of: (i) subjecting an aqueous composition comprising dry and shelled peas to a fermentation in the presence of one or more lactic acid bacteria, preferably one or more species of lactobacilli, until the pH of said peas is between 3.5 and 5.5, measured at ambient temperature over 1 g of said peas which have been ground and then suspended in 9 g of water, preferably in which the pH of the dry peas before being fermented is at least 6.0, measured at room temperature over 5 g of dry peas which were ground with 95 g of water; (ii) grinding said peas; (iii) fractionating said milled peas to obtain at least one protein fraction, optionally simultaneously with step (ii), by adjusting the pH of the ground peas to a pH of at least 6.0, for example between 6 , 0 and 9, preferably between 7 and 9; preferably, this pH adjustment is carried out on an aqueous composition comprising ground peas having a dry matter content of at most 45%, preferably at most 40%, preferably at most 35%, preferably not more than 30%, preferably not more than 25%; in one embodiment, the dry matter content of the ground peas is adjusted to the above-mentioned dry matter content by adding water accordingly; (iv) isolating the pea proteins from the at least one protein fraction.
In a preferred embodiment, the present invention relates to a method of extracting pea proteins of the species Pisum sativum, comprising the steps of: (i) subjecting an aqueous composition comprising dry and shelled peas to a fermentation in the presence of one or more lactic acid bacteria, said dried hulled peas having a dry matter content of 80% to 95% based on the total weight of the dried hulled peas; preferably in the presence of at least one or more species of lactobacilli until the pH of said peas is between 3.5 and 5.5, measured at ambient temperature over 1 g of said peas which have been ground then put into suspension in 9 g of water, preferably in which the pH of the peas before being fermented is between 6.0 and 7.0, measured at room temperature over 5 g of dry peas which have been ground with 95 g of water; (ii) grinding said peas; (iii) fractionating said ground peas to obtain at least one protein fraction, optionally simultaneously with step (ii), by adjusting the pH of said ground peas to a pH of at least 6.0, for example between 6 , 0 and 9.0, preferably between 7.0 and 9.0; preferably, this pH adjustment is carried out on an aqueous composition comprising ground peas having a dry matter content of at most 45%, preferably at most 40%, preferably at most 35%, preferably not more than 30%, preferably not more than 25%; in one embodiment, the dry matter content of the ground peas is adjusted to the above-mentioned dry matter content by adding water accordingly; (iv) isolating the pea proteins from the at least one protein fraction.
In a more preferred embodiment, the present invention relates to a process for extracting pea proteins of the species Pisum sativum, comprising the steps of: (i) subjecting an aqueous composition comprising dry and shelled peas to fermentation in the presence of one or more lactic acid bacteria at a temperature between 35 ° C and 45 ° C until the pH of said peas is between 3.5 and 5.5, measured at room temperature over 1 g of said peas which were crushed and then suspended in 9 g of water, wherein said dry husked peas have a dry matter content of 80% to 95% based on the total weight of dry husked peas; preferably in the presence of one or more species of lactobacilli; preferably in which the pH of the peas before being fermented is between 6.0 and 7.0, measured at ambient temperature over 5 g of dry peas which have been ground with 95 g of water, measured on the aqueous composition comprising peas, after said composition has been milled; (ii) grinding said peas; (iii) fractionating said ground peas to obtain at least one protein fraction, optionally simultaneously with step (ii), by adjusting the pH of said ground peas to a pH of at least 6.0, for example between 6 , 0 and 9, preferably between 7 and 9; preferably, this pH adjustment is carried out on an aqueous composition comprising ground peas having a dry matter content of at most 45%, preferably at most 40%, preferably at most 35%, preferably not more than 30%, preferably not more than 25%; in one embodiment, the dry matter content of the ground peas is adjusted to the above-mentioned dry matter content by adding water accordingly; (iv) isolating the pea proteins from the at least one protein fraction.
In a most preferred embodiment, the method for extracting pea proteins of the species Pisum sativum comprises the steps of: (i) subjecting an aqueous composition comprising dry and shelled peas to a fermentation in the presence of one or more lactic acid bacteria at a temperature between 35 ° C and 45 ° C until the pH of said peas is between 3.5 and 5.5, measured at room temperature over 1 g of said peas that have been crushed and then suspended in 9 g of water, said dried hulled peas having a dry matter content of 80% to 95% based on the total weight of dried hulled peas; preferably in the presence of one or more species of lactobacilli; the pH of the peas before being fermented being preferably between 6.0 and 7.0, measured at ambient temperature over 5 g of dry peas which have been ground with 95 g of water; (ii) grinding said peas; (iii) fractionating said ground peas to obtain at least one protein fraction, optionally simultaneously with step (ii), by adjusting the pH of said ground peas to a pH of at least 6.0, for example between 6 , 0 and 9.0, preferably between 7.0 and 9.0; preferably, this pH adjustment is carried out on an aqueous composition comprising ground peas having a dry matter content of at most 45%, preferably at most 40%, preferably at most 35%, preferably not more than 30%, preferably not more than 25%; in one embodiment, the dry matter content of the ground peas is adjusted to the above-mentioned dry matter content by adding water accordingly; (iv) isolating the pea proteins from the at least one protein fraction; (v) obtaining said isolated pea proteins as an aqueous suspension; (vi) optionally subjecting said aqueous suspension to at least one heat treatment.
In a preferred embodiment, steps (iii) to (vi) of the above method comprise the following steps: (1) fractionating said ground peas by separating a fraction comprising proteins from an insoluble fraction, preferably by decantation ; (2) subjecting the protein fraction to heat treatment at a temperature of between 50 ° C and 80 ° C; (3) precipitating the proteins included in said protein fraction by isoelectric precipitation, preferably by adjusting the pH of said fraction to a value between 4.5 and 5.8; preferably, this pH adjustment is carried out on an aqueous protein fraction having a solids content of at least 1%, preferably a solids content of between 3% and 10%; (4) isolating the precipitated proteins, preferably by decantation; thereby obtaining said isolated pea proteins as an aqueous suspension; (5) adjusting the pH of the aqueous suspension to a value between 6.0 and 8.0, or alternatively adjusting the pH of the aqueous suspension to 4.5 to 5.8; preferably, this pH adjustment is carried out on an aqueous suspension having a dry matter content of at most 45%, preferably at most 40%, preferably at most 35%, preferably at most 30%, preferably at most 25%, and in one embodiment the solids content can be adjusted to this extent by dilution with water; (6) optionally subjecting the pH adjusted aqueous suspension to heat treatment at a temperature of at least 70 ° C, preferably at least 75 ° C, preferably at a temperature of between 75 ° C and 210 ° C ° C, preferably between 85 ° C and 160 ° C, for example between 90 ° C and 150 ° C, for example between 95 ° C and 140 ° C, preferably for a period between 0.01 s and 10 min ; and (7) drying the aqueous suspension.
In a preferred embodiment, steps (iii) to (vi) of the above method comprise the following steps: (1) fractionating said ground peas by separating a fraction comprising proteins from an insoluble fraction, preferably by decantation ; (2) subjecting the protein fraction to heat treatment at a temperature of between 50 ° C and 80 ° C; (3) precipitating the proteins included in said protein fraction by isoelectric precipitation, preferably by adjusting the pH of said fraction to a value between 4.5 and 5.0; preferably, this pH adjustment is carried out on an aqueous protein fraction having a solids content of at least 1%, preferably a solids content of between 3% and 10%; (4) isolating the precipitated proteins, preferably by decantation; thereby obtaining said isolated pea proteins as an aqueous suspension; (5) adjust the pH of the aqueous suspension to a value between 6.0 and 8.0, preferably this pH adjustment is carried out on an aqueous suspension having a solids content of not more than 45%, of preferably at most 40%, preferably at most 35%, preferably at most 30%, preferably at most 25%, and in one embodiment the dry matter content can be adjusted in this measurement by dilution with water; (6) optionally subjecting the pH adjusted aqueous slurry to heat treatment at a temperature of from 90 ° C to 150 ° C, preferably for a period of from 0.01 s to 10 min; (7) drying the aqueous suspension to thereby obtain said pea proteins.
In a preferred embodiment, steps (iii) to (vi) of the above method comprise the following steps: (1) fractionating said ground peas by separating a fraction comprising proteins from an insoluble fraction, preferably by decantation ; (2) subjecting the protein fraction to heat treatment at a temperature of between 50 ° C and 80 ° C; (3) precipitating the proteins included in said protein fraction by isoelectric precipitation, preferably by adjusting the pH of said fraction to a value between 4.5 and 5.8; preferably, this pH adjustment is carried out on an aqueous protein fraction comprising at least 1.0% dry matter on the basis of the total weight of the composition, preferably at least 2.0% of dry matter, more preferably at least 3.0% dry matter, such as for example at least 4.0% dry matter, such as for example at least 5.0% dry matter; preferably between 1.0% and 40% of dry matter, preferably between 2.0% and 30% of dry matter, more preferably between 3.0% and 20% of dry matter, more preferably between 3.0% and 15% dry matter, such as between 3.0% and 10%; (4) isolating the precipitated proteins, preferably by decantation; thereby obtaining said isolated pea proteins as an aqueous suspension; (5) adjusting the pH of the aqueous suspension to 4.5 to 5.8; preferably, this pH adjustment is carried out on an aqueous suspension having a dry matter content of at most 45%, preferably at most 40%, preferably at most 35%, preferably at most 30%, preferably at most 25%, and in one embodiment the solids content can be adjusted to this extent by dilution with water; (6) optionally subjecting the pH adjusted aqueous suspension to heat treatment at a temperature of from 90 ° C to 150 ° C, preferably for a period of 10.0 to 10 minutes; and (7) drying the aqueous suspension to thereby obtain said pea proteins.
The pea proteins obtained using the methods according to the invention described here have different characteristics, such as different biochemical and / or organoleptic characteristics, as well as a difference in the values of the parameters related to the quality compared with the pea proteins known in the prior art.
Accordingly, the present invention also encompasses pea proteins, pea protein extracts, and pea protein compositions obtained or obtainable by the methods of the invention described herein. Those skilled in the art will appreciate that when reference is made to "pea proteins" in some embodiments, a composition is effectively described that includes primarily, but not exclusively, pea proteins. Residual impurities may be present in such compositions. Such residual impurities may include, for example, minerals, sugars, etc. As used herein, the term "pea protein" refers preferably to a pea protein extract or a composition comprising (on a dry matter basis) at least 70% by weight of proteins, preferably at least 80% by weight of proteins, more preferably at least 85% by weight of proteins. Preferably, the term "pea protein" refers to a composition comprising (on the basis of dry matter) from 70% by weight to 98% by weight of proteins, preferably from 80% by weight to 98% by weight of proteins, more preferably from 85% by weight to 98% by weight of proteins, more preferably from 88% by weight to 98% by weight of proteins.
In another aspect, the present invention relates to a composition comprising pea proteins obtained or obtainable using the methods of the invention described herein. In a preferred embodiment, such a composition is an edible composition. Preferably, said composition is a food intended for human or animal food, more preferably a dairy product, a confectionery product, a beverage, a meat product, a vegetarian product, a food supplement, a nutritional product intended for the control of athletes, a food for medical use, a food for the elderly or a bakery product. In a preferred embodiment, said food for human consumption is a biscuit, a bread, a cake, a waffle, or a soft caramel.
Accordingly, in another aspect, the present invention relates to the use of pea proteins described herein, in particular pea proteins obtained or obtainable by the methods described herein, in products intended for human or animal consumption. . In a preferred embodiment, the food products are selected from the group consisting of dairy products, confectionery products, beverages, meat products, vegetarian products, dietary supplements, nutritional products for weight control and sports foods, medical foods, senior citizens' foods and bakery products. In a preferred embodiment, the food products are bakery products or confectionery products for human consumption. The pea proteins described herein may, for example, partially or completely replace other proteins in products intended for human or animal consumption, for example proteins of animal origin, such as milk proteins. Particularly suitable applications of the pea proteins described herein may, for example, involve applications in which the Maillard reaction occurs, ie browning or icing reactions, such as those generally present in the preparation processes of bakery products or confectionery products.
Aspects and embodiments of the invention are further supported by the following non-limiting examples.
EXAMPLES
protocols
Unless otherwise indicated, in the examples below, all parameters are measured as defined in this part. The measurement of the parameters, as defined in this part, also represents in the preferred embodiments the method of measuring said parameters according to the invention as indicated in the respective aspects and embodiments of the detailed description above.
PH measurement on dry peas or an aqueous composition comprising ground peas or peas
The pH was measured using a WTW SERIES Inolab Termil 740 pH meter. The instrument was calibrated using pH 4.01 buffer solutions (WTW pH 4.01 technical buffer, STP4 model). , Order No. 108706) and pH 7 (WTW Technical Buffer pH 7.00, Model STP7, Order No. 108708).
When the pH was measured on the pea-free aqueous composition, a sample of aqueous solution was taken directly from the fermentation tank. The pH of the sample was measured once the value stabilized.
When the pH was measured on peas, the peas were taken from the fermentation tank. The peas were drained in a strainer and then placed on an absorbent paper for 2 minutes to remove excess juice. The peas were milled for 1 min using a mixer (Magic Bullet, Homeland Housewares). 1 g of ground peas was suspended in 9 g of deionized water (water conductivity <15 μS). The suspension was then crushed again using the mixer. Finally, the pH of the suspension (at room temperature) was measured once the value stabilized.
When the pH was measured on dry peas, the peas were milled dry for 1 min using a blender (Kenwood). 5 g of ground dry peas were suspended in 95 g of deionized water (water conductivity <15 μS). The suspension was then homogenized on a stir plate for 1 min. The pH of the suspension was measured once the value stabilized.
PH measurement on protein extract powder
The pH was measured using a pH meter WTW pH / Cond 340I / SET. The instrument was calibrated with buffer solutions at pH 4.01 (WTW pH 4.01 technical buffer, model STP4, order number 108706) and at pH 7 (WTW pH 7.00 technical buffer, model STP7 , Order No. 108708). 5.0 g of protein extract powder was introduced into a 100 ml beaker and supplemented to 50 g (Ohaus balance ARC120, sensitivity 0.01 g, capacity 3100 g) with deionized water at room temperature . The suspension was stirred for 5 min on a stirring plate (Stuart US151) set at speed 4. The pH of the suspension was measured (at room temperature) with stirring once the value stabilized.
PH measurement of food products
The pH meter (Knick Portavo 902 PH) was calibrated with buffer solutions at pH 4.01 (WTW Technical Buffer pH 4.01, Model STP4, Order No. 108706) and pH 7 (WTW pH Technical Buffer). 7.00, STP7 model, order No. 108708). The pH was measured by introducing the pH meter probe (Knick Portavo 902 PH) directly into the product (liquid food product, paste ...) at room temperature. In the case of a solid food product, a 50% dilution in demineralized water was performed and the solution was analyzed. After stabilization, the pH value was noted. Enumeration of lactic acid bacteria
Dilutions of the samples were carried out with EPT Dilucups 9 ml Led techno.
The medium used was MRS agar (according to DE MAN, ROGOSA and SHARPE) marketed by Merck Cat. No. 1.10661.0500.
The peas or the pea suspension were crushed using a grinder, Magic Bullet, Homeland Housewares.
When analyzing a sample of the pea-free aqueous composition, a sample was taken directly from the fermentation tank. 1 ml of sample was spread. When dilution was necessary, 1 ml of sample was added to the dilution cup and this step was repeated until the correct dilution was obtained, and then 1 ml of the diluted sample was spread. Petri dishes were incubated for 48 h at 45 ° C.
When analyzing a sample of peas, whole peas were taken from the fermentation tank. The peas were drained in a strainer and then placed on an absorbent paper for 2 minutes to remove excess juice. The peas were ground for 1 min. The ground peas were suspended (1 g pea in 9 g deionized water) in deionized water (conductivity <15 μS). The suspension was then milled using the mixer. 1 ml of suspension was spread. When dilution was necessary, 1 ml of suspension was added to the dilution cup and this step was repeated until the correct dilution was obtained, then 1 ml of the diluted sample was spread. Petri dishes were incubated for 48 h at 45 ° C. Determination of the dry matter content
The total solids content was determined gravimetrically as the residue content remaining after drying. Moisture was evaporated from the sample by drying in an oven. 5 g of sample were weighed in a previously weighed dry aluminum cup (Ohaus precision balance, capacity 410 g, sensitivity 0.001 g). The sample was placed in an oven at 103 ° C until the residual weight remained constant (at least 24 h). The sample was cooled in a desiccator for 1 h and 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 dry matter content of food products
The dry matter content of the food products was determined in duplicate after desiccation of 5 g of sample at 104 ° C overnight. Determination of the protein content by the Dumas method The apparatus (Leco FP2000) was calibrated with EDTA sold by Leco under the reference 502092. The quantities of EDTA weighed for carrying out the calibration were between 0.08 g and 0.50 g (0.08 g, 0.15 g, 0.25 g, 0.35 g, 0.40 g, 0.50 g). 0.3 g to 1 g of sample was weighed using a precision balance (Sartorius BP61 S, capacity 61 g, sensitivity 0.1 mg) and placed in a ceramic crucible. The ceramic crucible was automatically placed in an oven at 1200 ° C in which the sample was calcined in a combustion tube by pyrolysis under controlled oxygen flow. Nitrogen compounds were converted to N2 and NOx while other volatile decomposition compounds were retained by passage through adsorbent filters and a series of purification reagents. All nitrogen compounds were reduced to molecular nitrogen, which was quantitatively determined using a thermal conductivity detector. The nitrogen content was then calculated using a microprocessor.
The results are expressed as percentage of proteins (% N x 6.25):% nitrogen = g of nitrogen per 100 g of sample% protein =% nitrogen x 6.25 Determination of the nitrogen content in the ISA samples by Dumas method The apparatus (Leco FP2000) was calibrated with a glycine solution at 15 mg / ml (glycine powder marketed by Merck under the reference 1.04201.1000). The amounts of glycine solution at 15 mg / ml weighed for performing the calibration were between 0.1 g and 1.8 g (0.1 g, 0.4 g, 0.7 g, 1.1 g g, 1.4 g, 1.8 g). 1 g to 1.8 g of sample was weighed using a precision balance (Sartorius BP61S, capacity 61 g, sensitivity 0.1 mg) and placed in a ceramic crucible covered with a silicone insert. nickel. The ceramic crucible was automatically placed in an oven at 1200 ° C in which the sample was calcined in a combustion tube by pyrolysis under controlled oxygen flow. Nitrogen compounds were converted to N2 and NOx while other volatile decomposition compounds were retained by passage through adsorbent filters and a series of purification reagents. All nitrogen compounds were reduced to molecular nitrogen, which was quantitatively determined using a thermal conductivity detector. The nitrogen content was then calculated using a microprocessor.
The results are expressed as a percentage of nitrogen:% nitrogen = g of nitrogen per 100 g of sample. Determination of the solubility index of nitrogen (ISA)
After dispersing the proteins in deionized water, the solubility index of nitrogen was determined by measuring the ratio of the percentage of nitrogen in the supernatant after centrifugation to the percentage of nitrogen in the starting slurry. The method was used on a protein extract powder having a solids content of 90% to 99% (by weight) and was carried out in the month following the drying of the protein extract. The measurement was performed at room temperature. 9.0 g of sample were introduced into a 400 ml beaker and supplemented to 300 g (Ohaus ARC120 balance, 0.01 g sensitivity, 3100 g capacity) with deionized water at room temperature. The suspension was homogenized with a spoon and then stirred for 5 min on a stirring plate (Stuart US151) set at speed 4. 10 ml of the starting suspension was collected and the nitrogen content was adjusted. was analyzed using a Leco FP 2000 protein analyzer. The suspension was divided into two 150 ml beakers, the pH was increased in one and decreased in the other. The pH of the suspension was adjusted to pH 3.5, 4.5, 5.5, 6.5, 7 and 8 with 1 N HCl or 1 N NaOH (pH meter WTW pH / Cond 340I). /SET). For each pH adjustment, the pH value was recorded once stabilized and 10 ml of the suspension was collected in a 10 ml centrifuge tube. Aliquots of the suspension at different pH were centrifuged for 15 min at 6000 rpm (ALC 4239 R centrifuge). The various supernatants were collected and the nitrogen content was analyzed using a Leco FP 2000 protein analyzer. For each pH tested, the solubility index of nitrogen was calculated according to the following formula: % solubility index of nitrogen =% nitrogen in the supernatant /% nitrogen in the starting solution x 100 Determination of the isoelectric pH of the protein fraction 300 g of protein fraction having a protein content of 1% by weight based on of the total weight of the protein fraction were introduced into a 400 ml beaker at room temperature. The suspension was stirred for 5 min on a stirring plate (Stuart US151) set at speed 4. 10 ml of the starting suspension was collected and the nitrogen content was analyzed using an analyzer. Leco FP 2000 protein. The suspension was divided into two beakers of 150 ml, the pH was increased in one and decreased in the other. The pH of the suspension was adjusted to pH 3.5, 3.75, 4.0, 4.25, 4.5, 4.75, 5.0, 5.25, 5.5, 5.75, 6.0, 6.25, 6.5, 6.75, and 7.0 with 1N HCl or 1 N NaOH (pH meter WTW pH / Cond 340I / SET). For each pH adjustment, the pH value was recorded once stabilized and 10 ml of the suspension was collected in a 10 ml centrifuge tube. Aliquots of the suspension at different pH were centrifuged for 15 min at 6000 rpm (ALC 4239 R centrifuge). The various supernatants were collected and the nitrogen content was analyzed using a Leco FP 2000 protein analyzer. For each pH tested, the solubility index of nitrogen was calculated according to the following formula: % solubility index of nitrogen =% nitrogen in the supernatant /% nitrogen in the starting solution x 100
The isoelectric pH was determined as the pH at which the solubility index of nitrogen was the lowest. Determination of sugar content The sample was prepared with an Eppendorf 5417R centrifuge and with NANOSEP 100k OMEGA centrifugal devices.
The peas or pea suspension were ground using a blender, Magic Bullet, Homeland Housewares.
When analyzing a sample of the pea-free aqueous composition, a sample was taken directly from the fermentation tank. The sample was diluted 20-fold (1 g pea juice in 19 g deionized water) with deionized water (conductivity <15 μS). 0.5 ml of this dilution was placed in an Eppendorf filter and centrifuged at 14000 rpm for 10 min. The filtrate was then used to analyze the sugar content.
When preparing a pea sample, whole peas were taken from the fermentation tank. The peas were drained in a strainer and then placed on an absorbent paper for 2 minutes to remove excess juice. The peas were ground for 1 min. The ground peas were suspended (1 g pea in 9 g deionized water) in deionized water (conductivity <15 μS). The suspension was then milled using the mixer.
The suspension was diluted 8 times (1 g pea suspension in 8 g deionized water) with deionized water (conductivity <15 μS). 0.5 ml of this dilution was placed in an Eppendorf filter and centrifuged at 14000 rpm for 10 min. The filtrate was then used to analyze the sugar content.
A Thermo scientific - Dionex ICS 5000 chromatographic system with Chroneleon 6.80 SR11 Build 3161 software was used for the analysis of sugars. The separation was carried out using a Carbopac PA100 column 4 mm x 250 mm (+ protection) at 40 ° C. Elution was carried out with 40 mM NaOH at a flow rate of 1 ml / min. The injection volume was 10 μl. Four-pulse detection mode was used for pulse amperometric detection (PAD). Calibration was performed using appropriate standard solutions for each of the following sugars:
The concentrations of the standard solutions (Et, 1, 2, 3 and 4) (mg / l) are shown in the table below.
Acidity measurement Acidity was measured using a WTW SERIES Inolab Termil 740 pH meter. The instrument was calibrated using pH 4.01 buffer solutions (WTW pH Technical Buffer). 4.01, model STP4, order No. 108706) and pH 7 (WTW technical buffer pH 7.00, model STP7, order No. 108708).
The peas or pea suspension were ground using a blender, Magic Bullet, Homeland Housewares.
When measuring the acidity of "pea juice", a sample (A) was taken directly from the fermentation tank. Sample (A) was weighed. A solution of sodium hydroxide at 1 mol / l (C) (No. 1.09137.1000 TitriPURR, density = d = 1.04 kg / l) was added slowly until the pH of the sample was stabilized at pH 7 for at least 2 min. The mass of sodium hydroxide (B) was then calculated. acidity (mEq / kg) = (B x (C / d) / A) x 1000
When measuring the acidity of the peas, whole peas were taken from the fermentation tank. The peas were drained in a strainer and then placed on an absorbent paper for 2 minutes to remove excess juice. The peas were ground for 1 min. The ground peas were suspended (1 g pea in 9 g deionized water) in deionized water (conductivity <15 μS). The suspension was then milled using the mixer. A pea suspension was obtained.
The exact amount of the pea suspension (A ') was weighed. A solution of 1 mol / l sodium hydroxide (C ') (No. 1.09137.1000 TitriPURR, density = d = 1.04 kg / l) was added slowly until the pH of the suspension was stabilized at pH 7 for at least 2 min. The mass of sodium hydroxide (B ') was then calculated. acidity (mEq / kg) = (B 'x (C / d) / (A7 10)) x 1000 Determination of ash content
The ash content was determined gravimetrically as the residue content remaining after heating in a high temperature muffle furnace. Moisture was evaporated from the sample by drying in an oven. 2 g of sample were weighed in a previously weighed dry porcelain crucible (Ohaus precision balance, capacity 410 g, sensitivity 0.001 g). The crucible was placed in a muffle furnace at 550 ° C for 24 hours. The crucible was placed for 1 hour in an oven at 103 ° C. and then in a desiccator for 1 hour. After cooling, the crucible was weighed. The results are expressed in% (g of ash per 100 g of sample). ash (%) = (m3 - m1) / (m2-m1) x 100 m1 = weight of the crucible (in g) m2 = weight of the crucible with the sample (in g) m3 = weight of the crucible with the ashes (in boy Wut)
Determination of potassium content by ICP-AES
The determination of the potassium content was carried out by ionization of the sample in an inert gas plasma. The ICP-AES (Inductively Coupled Plasma Atomic Emission Spectroscopy) apparatus was calibrated using Merck Chloride marketed under the reference 104938. The weight of potassium chloride used for calibration was adapted according to the potassium content of the sample. 2 g of ash were prepared from the sample according to the method of determining the ash content. The ashes were diluted in deionized water so as to be in the reading range of the apparatus. The solution was filtered on Whatman 595 1/2 185 mm paper. The filtered sample was ionized by injection into the ICP-AES. The results are expressed in mg / kg or ppm (mg of potassium per kg of sample).
Determination of magnesium content by ICP-AES
The determination of the magnesium content was carried out by ionizing the sample in an inert gas plasma. The ICP-AES (inductively coupled plasma atomic emission spectrometry) apparatus was calibrated with ICP standard magnesium sold by Merck as 170331 (1000 mg / l) or 170379 (10000 mg / l) . The magnesium standard ICP weight used for calibration was adapted according to the magnesium content of the sample. 2 g of ash were prepared from the sample according to the method of determining the ash content. The ashes were diluted in deionized water so as to be in the reading range of the apparatus. The solution was filtered on Whatman 595 1/2 185 mm paper. The filtered sample was ionized by injection into the ICP-AES. The results are expressed in mg / kg or ppm (mg of magnesium per kg of sample).
Determination of viscosity using a Brookfield DVII viscometer
The determination of the viscosity of a protein suspension using a Brookfield DVII viscometer consists in measuring its resistance to flow imposed by the rotation of a cylindrical probe. This resistance causes the torsion of a spring attached to the detector of a drive system. The value of the viscosity, expressed in centipoise (cP), is proportional to the percentage of torsion indicated by the viscometer and to a multiplying factor depending on the probe used and its rotational speed. The method was used on a protein extract powder having a solids content of 90% to 99% (by weight) and was carried out in the month following the drying of the protein extract. The measurement was performed at room temperature.
A suspension containing 13.5% protein (by weight) was prepared. 75 g of sample were weighed (Ohaus balance ARC120, sensitivity 0.01 g, capacity 3100 g) in a beaker of 250 ml and the necessary quantity of demineralized water was weighed in a plastic beaker of 1 I, the two weighings having been carried out at ambient temperature. The powder was suspended in water with mechanical stirring (IKA, EURO-ST.P CV) at 700 rpm for 5 minutes using a dissolver 80 cm in diameter (marketed by Roth under A322.1). The pH of the suspension was measured with stirring (pH meter WTW pH / Cond 340I / SET). Stirring was stopped for 3 min and the viscosity of the slurry was measured at three different locations using a Brookfield DVII + Pro viscometer at a speed of 50 rpm. The probe used for the measurement was chosen between SOI and SO7 so that the percentage of torsion was between 20% and 80%. The value of the viscosity was recorded after 4 s of rotation of the probe. The suspension was again mechanically stirred for 5 min at 700 rpm during which the pH was adjusted to 6.4 with 3N HCl. Stirring was stopped for 3 min and the viscosity of the suspension was measured in the same manner as before. Similarly, the viscosity of the suspension was measured at pH 6.2, 6.0 and 5.8 after stirring for 5 minutes at 700 rpm and 3 min of rest.
Once the initial pH of the suspension containing 13.5% protein reached 5.8 or lower, the pH was raised to pH 7.5 with 3N NaOH, instead of being decreased with 3N HCI
Color measurement
The L * a * b * coordinates were measured at 20 ° C using a CR5 chromameter (Konica Minolta TA Sensing, Europe). L * is the brightness on a scale of 0 to 100 from black to white; a *, (+) red or (-) green; and b *, (+) yellow or (-) blue.
Device: - CR5 chromameter (Konica Minolta TA Sensing Europe). - Petri dish CR-A502 Method: Sample preparation - The Petri dish was filled with the sample so that the analysis was done on a uniform surface.
Process: - the petri dish was placed on the apparatus at the specifically reserved place and the analysis was started Results: - the L * a * b * values are indicated by the chromameter (average of 3 measurements). Sensory analysis of proteins in solution The sensory evaluation was carried out by a group of 5 members trained for this purpose. The formation of the members of the group was based on the recognition of six characteristics (sweetness, bitterness, metallic taste, salty taste, acidity, umami taste and astringency). Descriptive analysis was performed on 4% dispersions. After discussion to reach agreement, the descriptive terms that were most important to characterize the appearance, texture and taste of the solutions were selected.
Water Activity Water activity is a measure of the energy status of water in a system. It is defined as the vapor pressure of water in a substance divided by that of pure water at the same temperature. The pure distilled water therefore has an activity of exactly 1. The determination of the water activity (Ae) was carried out using a Rotronic Hygroskop DT apparatus, Krautli.
A cell was filled with the sample to be characterized and placed in the measuring chamber (Rotronic Hygroskop DT, Krautli). After stabilization, the value of the water activity was recorded.
Sensory analysis of cooked products The sensory evaluation was carried out by a group of 5 members trained for this purpose. The formation of the members of the group was based on the recognition of six characteristics (sweetness, bitterness, metallic taste, salty taste, acidity, umami taste and astringency). A descriptive analysis was performed on the finished products. After discussion to reach agreement, the descriptive terms that were most important to characterize the appearance, texture and taste of the products were selected.
Hardness of biscuits
The hardness of biscuits is defined as the force required to break a biscuit with a knife. The hardness of the biscuits was evaluated using a Ta-XT2i texture analyzer.
Device: - TA-XT2I Texture Analyzer (Stable Micro Systems, Ltd) - Compression Sensor, 25 kg - Knife Set with Knife (HDP / BSK) Method: - Position the limit of the top rail so that the Warner blade Bratzler is 1 mm above the surface of the sample - set the device TA-XT2Ï: o measure the force in Compression - go back to the beginning o carry out a pre-test of the speed: 3 mm / so test the speed: 2 mm / so proceed to post-test speed: 10 mm / so penetration distance: 5 mm o trigger type: auto - 3 g - start of the penetration test. The results were recorded using a texture analyzer and plotted on a graph. Results
The hardness of the biscuits was the maximum force recorded during the test (expressed as "max force"). The test results were obtained from 20 samples and the average value was calculated.
EXAMPLE 1 Method for Extraction of Pea Proteins According to an Embodiment of the Present Invention
This example was carried out following the protocol shown schematically in FIG.
Peas harvested dry, referred to herein as "dry peas" (having a dry matter content (based on the total weight of dry peas) of about 87%) were sieved and spiked by passage through a cleanser. Then, the peas were shelled in a dehuller.
Then the peas were fermented with lactic acid bacteria (Lactobacillus fermentum). To this end, the peas have been soaked in drinking water in a discontinuous manner. In subsequent batches, a portion of the fermentation medium (pea-free aqueous phase) of a previous batch was used as an inoculum for subsequent fermentation. The peas were fermented in the presence of about 10 8 cfu of lactic acid bacteria per ml of aqueous composition comprising peas. 400 kg of peas per m3 of the total volume of aqueous composition comprising peas were placed in a tank. The fermentation was carried out under anaerobic conditions in a closed vessel without degassing at a temperature of about 40 ° C until a pea pH of 4.4 was obtained. During the fermentation, the aqueous phase in the fermentation tank was recycled to about 20 m3 / h. The peas were fermented for 480 minutes. At the end of the fermentation, the peas absorbed a quantity of water equal to about their initial mass before being fermented and had a solids content of about 43% (by weight).
After fermentation, the peas were removed from the fermentation medium. The peas were then placed in a perforated rotating drum and washed to remove the remaining fermentation medium. After cleaning, the peas were subjected to wet milling. During grinding, additional drinking water was added so that the final composition had a solids content of about 25% (by weight). During the milling step, the pH was adjusted to about 8 by adding sodium hydroxide.
After milling and pH adjustment, the ground pea paste was decanted by centrifugation. The protein-containing supernatant and soluble impurities had a solids content of about 4% (by weight).
The aqueous protein fraction was then subjected to heat treatment at 75 ° C for 15 seconds in a plate heat exchanger.
Then, the pea proteins were isolated by isoelectric precipitation. For this purpose, the pH of the fraction containing the pea proteins was adjusted to 4.7 with sulfuric acid. The separation of the precipitated / aggregated proteins was carried out by decantation by centrifugation. The fraction containing the resulting pea protein (aqueous suspension) had a solids content of about 25% (by weight). Drinking water was added until a solids content of 14% (by weight) was obtained.
Then, the pH of the aqueous suspension was adjusted to 7.6 with sodium hydroxide. The aqueous suspension was then heat treated by heating to about 90 ° C using a plate heat exchanger, and maintaining the slurry at a temperature of about 90 ° C for 7 min.
Finally, the suspension was spray dried. The inlet temperature of the spray dryer was about 150 ° C and the exit temperature was about 70 ° C.
Example 2 Evolution of the sugar content and the pH / acidity of the peas during the fermentation step of a method according to one embodiment of the invention
This example was carried out following the protocol described below. The experiment was repeated about 65 times with different fermentation times.
Peas harvested dry, referred to herein as "dry peas" (having a dry matter content (on the basis of the total weight of dry peas) of about 87%) were sieved and peeled by passage through a cleanser. Then, the peas were shelled in a dehuller.
Then the peas were fermented with lactic acid bacteria (Lactobacillus fermentum). To this end, the peas have been soaked in drinking water in a discontinuous manner. In subsequent batches, a portion of the fermentation medium (pea-free aqueous phase) of a previous batch was used as an inoculum for subsequent fermentation. The peas were fermented in the presence of 108 ml of lactic acid bacteria per ml of aqueous composition comprising peas. 400 kg of peas per m3 of the total volume of aqueous composition comprising peas were placed in a tank. The fermentation was carried out under anaerobic conditions in a closed vessel without degassing at a temperature of about 40 ° C. During the fermentation, the aqueous phase in the fermentation tank was recycled to about 20 m3 / h. The peas were fermented for different durations as summarized in Figure 2. After fermentation, the peas were removed from the fermentation medium and subjected to various analyzes.
The sugar content of dried peas that were shelled before being fermented was calculated on 20 pea samples and averaged 8% by weight (based on dry matter), a minimum of 6.4% by weight, and at most 9% by weight. The sugar content was based on the total concentration of glucose, fructose, sucrose, verbascose, raffinose, stachyose, and galactose. The evolution of sugar content in peas during fermentation is shown in Figure 2.
The pH and acidity of the fermentation medium, as well as the pH and acidity of the peas, were evaluated during the fermentation. Figures 3 and 4 illustrate the evolution, respectively, pH and acidity inside the peas and in the fermentation medium (also referred to as the "juice").
Example 3 Evolution of Bacterial Development During the Spontaneous Fermentation Stage of a Method According to an Embodiment of the Invention
This example was carried out following the protocol described below. The experiment was repeated 7 times.
Peas harvested dry, referred to herein as "dry peas" (having a dry matter content (based on the total weight of dry peas) of about 87%) were sieved and spiked by passage through a cleanser. Then, the peas were shelled in a dehuller.
Then the peas were subjected to spontaneous fermentation with lactic acid bacteria. To this end, the peas have been soaked in drinking water in a discontinuous manner. No inoculum was used to carry out the fermentation. 400 kg of peas per m3 of the total volume of aqueous composition comprising peas were placed in a tank. Fermentation was performed under anaerobic conditions in a closed vessel without degassing at a temperature of about 40 ° C during Experiments 1 to 6 and at a temperature of about 45 ° C during Experiment 7 (lab kinetic ). During fermentation, the aqueous phase in the fermentation tank was recycled. The peas were fermented for between 100 min and 900 min as shown in Figure 5.
Figure 5 shows a graph plotting the concentration of lactic acid bacteria in the aqueous pea-containing composition contained in the first fermentation tanks of a series of experiments as a function of the fermentation time.
Example 4: Measurement of the pH of different waters, with or without the addition of husked ground peas
All pH values were measured at room temperature using a pH meter calibrated the day before measurements.
Table 1 illustrates the pH of the different types of water used: tap water, deionized water, and treated well water. Well water has been treated in a manner that is safe for human consumption in accordance with Directive 98/83 / EC (also referred to herein as "drinking water").
Table 1
Table 2 illustrates the pH of the shredded ground pea suspensions containing 25% by weight of dry matter. The pH was determined after suspending the ground peas in different types of water for 1 min with magnetic stirring (200 rpm).
Table 2
Table 3 illustrates the pH of the shredded ground pea suspensions in treated well water containing increasing concentrations of dry matter. The pH was determined after suspending the ground peas in water for 1 min with magnetic stirring (200 rpm).
Table 3
Table 4 illustrates the pH of the aqueous (pea-free) phases of dry shelled whole pea suspensions (270 g pea + 520 g water) in the indicated water types, which were then homogenized for 5 seconds. The pH was measured immediately after homogenization of peas and water.
Table 4
EXAMPLE 5 Comparison of Protein Extraction Processes with Fermentation (According to an Embodiment of the Invention) and Without Fermentation (Comparative Example) The Protein Extract 3 (Product 3) Was Prepared (According to the Invention) as described in Example 1 from dry husked peas using a process comprising a fermentation step (fermentation for 8 h at 40 ° C in the presence of Lactobacillus fermentum). The experiment was repeated using identical conditions and product 4 was obtained. The protein extract 1 (product 1) was prepared (not according to the invention) from dry husked peas that were not subjected to a fermentation step but only to a hydration step (hydration during 40 minutes). min at 15 ° C). The experiment was repeated using identical conditions and product 2 was obtained.
In all cases, the peas were hydrated, but to a lesser extent than in the non-fermentation method with a pea dry matter content of 67% (by weight). After hydration and / or fermentation, the peas were removed in all cases from the aqueous phase and subjected to wet grinding in the presence of additional drinking water so that the final composition had a solids content of about 24%. In the non-fermentation process, pea pH did not drop and was about 6.5; while in the fermentation process, pea pH was considerably reduced and reached a value of 4.4. After grinding, the extraction process was similar in both cases up to the milling stage and as described in Example 1. It was observed that the purity of the proteins after grinding was lower in the process without fermentation. On the other hand, the tendency of the heat exchangers to foul during the subsequent heat treatment was higher in the non-fermentation process. After milling, protein precipitation and the resulting solids content achieved similar efficiency in both the fermentation and the non-fermentation processes. In the final pea protein extracts, the purity of the proteins was lower in the process without fermentation. In addition, the total amount of sugars in the final pea protein extract was higher in the process without fermentation.
The differences in the physicochemical and functional properties of the proteins extracted with or without fermentation can be summarized as follows: the general composition was similar, although the protein extract obtained without inclusion of the fermentation step had a content of K + superior (3 times) and a higher mg2 + content (1.6 times) compared with the protein extract obtained with inclusion of the fermentation step
Table 5 summarizes the levels of potassium and magnesium (based on dry matter) given in ppm / dry matter (ppm / MS) of products 2 and 4.
Table 5
the purity of the proteins was decreased by 1.5% in the protein extracts prepared without the fermentation step (86.0% of proteins on the basis of the dry matter) in comparison with the protein extracts prepared with the fermentation step (87.3% protein based on dry matter) - sugar content was increased about 3-fold in prepared protein extracts without inclusion of the fermentation step (1.40 % of sugars on the basis of dry matter) compared with the protein extracts prepared with inclusion of the fermentation step (0.45% of sugars on the basis of the dry matter). The sugar content was based on the total concentration of glucose, fructose, sucrose, verbascose, raffinose, stachyose, and galactose. the viscosity was decreased by about 3.5 times and 2.5 times respectively at pH 7.8 and pH 6.4 in the protein extracts prepared with the fermentation step compared with the protein extracts prepared without the fermentation step (see also Figure 6).
The viscosity measured for each extract at different pH's is shown in Table 6 and the viscosity profile is shown in Figure 6.
Table 6
the color of the dry protein extract was slightly more pink / orange in the extracts prepared without the fermentation step compared with the extracts prepared with the fermentation step, on the basis of a visual observation; also the color of the dispersed protein extracts (solution in 4% by weight water) was slightly more orange in the extracts prepared without the fermentation step compared to the extracts prepared with the fermentation step. the taste of the dispersed protein extract (solution in 4% by weight tap water) was determined to be more bitter and astringent in the extracts prepared without the fermentation step compared with the prepared extracts with the fermentation step.
Example 6 Food Products Comprising Pea Proteins According to the Invention The inclusion of pea protein in various food products was evaluated. 1. Biscuits
Cookie dough has been prepared. A pea protein was prepared as described below.
Dry harvested peas, referred to herein as "dry peas" (having a dry matter content (by weight) of about 87%) were sieved and spiked by passage through a cleanser. Then, the peas were shelled in a dehuller.
Then the peas were fermented with lactic acid bacteria (Lactobacillus fermentum). To this end, the peas have been soaked in drinking water in a discontinuous manner. In subsequent batches, a portion of the fermentation medium (pea-free aqueous phase) of a previous batch was used as an inoculum for subsequent fermentation. The peas were fermented in the presence of 108 ml of lactic acid bacteria per ml of aqueous composition comprising peas. 400 kg of peas per m3 of the total volume of aqueous composition comprising peas were placed in a tank. Fermentation was performed under anaerobic conditions in a closed vessel without degassing at a temperature of about 40 ° C until a pea pH of 4.7 was obtained. During the fermentation, the aqueous phase in the fermentation tank was recycled to about 20 m3 / h. The peas were fermented for about 430 minutes. At the end of the fermentation, the peas absorbed a quantity of water equal to about their initial mass before being fermented and had a solids content of about 47% (by weight).
After fermentation, the peas were removed from the fermentation medium. The peas were then placed in a perforated rotating drum and washed to remove the remaining fermentation medium. After cleaning, the peas were subjected to wet milling. During grinding, additional drinking water was added so that the final composition had a solids content of about 25% (by weight). During the milling step, the pH was adjusted to about 8 by adding sodium hydroxide.
After milling, the ground pea paste was decanted by centrifugation. The protein-containing supernatant and soluble impurities (also referred to herein as "an aqueous composition comprising pea proteins") had a solids content of about 4% (by weight).
The aqueous composition comprising pea proteins was then heat-treated at 75 ° C for 15 seconds in a plate heat exchanger.
Then, the pea proteins were concentrated by isoelectric precipitation. For this purpose, the pH of the aqueous composition comprising pea proteins was adjusted to 4.8 with sulfuric acid. The separation of the precipitated / aggregated proteins was carried out by decantation by centrifugation. The resulting pea protein concentrate was obtained as an aqueous suspension having a solids content of about 25% (by weight).
The dry matter content of the aqueous suspension was adjusted to about 16% (by weight) after addition of water; then the pH of the suspension was adjusted with sodium hydroxide to a pH of about 7.4. The slurry was then subjected to heat treatment at a temperature of about 90 ° C for about 7 minutes; then spray dried to obtain a powder (pea protein A) having a solids content of about 95% (by weight).
The dough was prepared as shown in Table 7.
Table 7
An analysis of the dough is given in Table 8.
Table 8
2. Soft toffee bars
Soft caramel bar recipes are shown in Table 9. Table 9
The method of preparing the bars was as follows:
- melt the fat at 45 ° C in a water bath - mix the syrups and add the fat - mix the powders in a Hobart mixer - add the syrups and shake for a few minutes until a paste homogeneous - the dough was placed in a plastic bag and dispersed, then left to rest for a whole night - cut the bars and coat them with chocolate
PH, water activity (Ae), and hardness of soft caramel bars were measured over time (months) and the results are shown respectively in Tables 10, 11, and 12.
Table 10
Table 11
Table 12

权利要求:
Claims (18)
[1]
A method of extracting pea proteins, comprising the steps of: (a) fermenting an aqueous composition comprising peas, preferably in the presence of lactic acid bacteria; (b) grinding said peas; (c) fractionating said ground peas to obtain at least one protein fraction; and (d) isolating the pea proteins from the at least one protein fraction.
[2]
The method of claim 1, wherein said peas of step (a) are fermented until the pH of said peas is at most 5.5, preferably at most 5, 0, more preferably between 3.5 and 5, preferably at room temperature over 1 g of said peas which have been ground and suspended in 9 g of water.
[3]
The process according to claim 1 or 2, wherein said peas of step (a) are fermented until the pH of said peas is reduced by at least 1 pH unit, preferably at least 1.5 pH units, preferably at room temperature over 1 g of said peas which have been ground and then suspended in 9 g of water.
[4]
A process according to any one of claims 1 to 3, wherein step (a) comprises adding to dry peas and / or pea peels an aqueous solution, preferably dry peas having a content content between 80% and 95% based on the total weight of the dry peas.
[5]
The process according to any one of claims 1 to 4, wherein step (a) comprises fermenting said peas until they have a dry matter content of between 35% and 60% based on total weight of the peas.
[6]
The process according to any one of claims 1 to 5, wherein said peas after step (a) and before step (b) have a dry matter content of between 35% and 60% based on total weight of the peas.
[7]
The process according to any one of claims 1 to 6, wherein said peas of step (a) are fermented for at least 3 hours, preferably for at least 3 hours and not more than 24 hours.
[8]
Process according to any one of claims 1 to 7, wherein said peas of step (a) are subjected to fermentation at a temperature of between 30 ° C and 50 ° C, preferably between 35 ° C and 45 ° C.
[9]
The process according to any one of claims 1 to 8, wherein step (a) comprises fermenting said peas with one or more species of lactobacilli.
[10]
The method according to any one of claims 1 to 9, wherein said peas of step (a) are fermented in the presence of at least 102 cfu to 1010 cfu of lactic acid bacteria per ml of said aqueous composition. including peas.
[11]
The method according to any one of claims 1 to 10, wherein the fractionation of said ground peas in step (c) comprises separating at least a portion of the proteins included in the pea, preferably a fraction comprising less than 50% by weight of protein on the basis of the total dry matter content of said fraction.
[12]
The process according to any one of claims 1 to 11, wherein the fractionation of said ground peas in step (c) comprises adjusting the pH of the ground peas to a pH of at least 6, preferably at least 7, most preferably at a pH of at least 8 and at most 9.
[13]
The process according to any one of claims 1 to 12, wherein the fractionation of said ground peas in step (c) comprises: subjecting said ground peas to one or more decantation steps, preferably one or more decantation steps by centrifugation.
[14]
The method of any one of claims 1 to 13, wherein isolating the pea protein from said protein fraction of step (d) comprises concentrating said pea protein.
[15]
The method according to any one of claims 1 to 14, wherein isolating the pea proteins from said protein fraction of step (d), comprises at least one step of precipitation, flocculation, filtration, and / or chromatography.
[16]
Pea proteins obtainable by the process of any one of claims 1 to 15.
[17]
An edible composition, preferably a food or feed product, comprising the pea protein of claim 16.
[18]
18. Use of the pea protein according to claim 16 in products intended for human or animal food, preferably in dairy products, confectionery products, beverages, meat products, vegetarian products, food supplements. , nutritional products for weight control and sports, medical foods, foods for the elderly and bakery products.

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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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FR2889416A1|2005-08-05|2007-02-09|Roquette Freres|COMPOSITION OF PEAS PROTEINS|

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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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EP131933830|2013-11-18|

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EP13193383|2013-11-18|CN201480062906.4A| CN105828633B|2013-11-18|2014-11-18|Method for extracting pea protein|

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SI201431545T| SI3071045T1|2013-11-18|2014-11-18|Method for extracting pea proteins|

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US15/035,298| US11019835B2|2013-11-18|2014-11-18|Method for extracting pea proteins|

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PCT/EP2014/074940| WO2015071499A1|2013-11-18|2014-11-18|Method for extracting pea proteins|

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DK14809775.1T| DK3071045T3|2013-11-18|2014-11-18|Method for extraction of pea proteins|

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CA2929050A| CA2929050C|2013-11-18|2014-11-18|Method for extracting pea proteins|

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PCT/EP2014/074939| WO2015071498A1|2013-11-18|2014-11-18|Method for extracting pea proteins|

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CA2929054A| CA2929054A1|2013-11-18|2014-11-18|Method for extracting pea proteins|

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EP14815249.9A| EP3071046A1|2013-11-18|2014-11-18|Method for extracting pea proteins|

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EP14809775.1A| EP3071045B1|2013-11-18|2014-11-18|Method for extracting pea proteins|

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CN201480062690.1A| CN105764347B|2013-11-18|2014-11-18|Method for extracting pea protein|

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US15/035,261| US10390548B2|2013-11-18|2014-11-18|Method for extracting pea proteins|

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ES14809775T| ES2800475T3|2013-11-18|2014-11-18|Method for extracting pea proteins|