Classifications

• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
  • A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
  • A23J1/14—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
  • A23J1/148—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds by treatment involving enzymes or microorganisms
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23K—FODDER
  • A23K20/00—Accessory food factors for animal feeding-stuffs
  • A23K20/10—Organic substances
  • A23K20/142—Amino acids; Derivatives thereof
  • A23K20/147—Polymeric derivatives, e.g. peptides or proteins
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
  • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
  • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
  • A23L33/17—Amino acids, peptides or proteins
  • A23L33/185—Vegetable proteins
• • A—HUMAN NECESSITIES
  • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
  • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
  • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

• the invention relates to the field of plant proteins, in particular protein isolates from legumes, even more particularly protein isolates from peas having a low lipid content.
• the human daily protein requirements are between 12 and 20% of the food ration. These proteins are supplied both by products of animal origin (meat, fish, eggs, dairy products) and by products of plant origin (cereals, legumes, seaweed).
• animal proteins have many disadvantages, both in terms of their allergenicity, in particular concerning proteins obtained from milk or eggs, and in terms of the environment in relation to the harmful effects of intensive breeding.
• a drawback of certain vegetable proteins, in particular legume proteins, even more particularly pea proteins, is that they are not tasteless. They can therefore bring unpleasant flavors (in English “off-flavors”) in the foods in which they are incorporated. These tastes are frequently described by consumers as tastes of legumes (in English "beany”), peas or a bitterness.
• a known solution to this problem is to mask these unpleasant flavors by introducing chemical compounds such as flavors during the manufacturing process.
• this solution is often not satisfactory because it does not mask the unpleasant flavor but only reduce it slightly.
• a second disadvantage is that the food manufacturing process is then more expensive due to the addition of additional ingredients. In addition, more and more consumers are turning away from products containing chemical compounds in favor of healthier food.
• a more advantageous solution is to directly use a vegetable protein isolate having little or no unpleasant flavor.
• Some examples of methods for obtaining such isolates have already been described.
• application WO201 5/071498 describes an extraction process by wet grinding, combined with lactic acid fermentation, to extract a purified pea protein isolate.
• Another example in application WO2017 / 120597 describes a process for precipitation in the form of salts, combined with a specific washing of the proteins with a large volume of an aqueous solution at neutral pH.
• these methods are not satisfactory because they result in protein isolates still exhibiting a pea-like taste and bitterness.
• Lipids being the substrates of lipoxygenase and oxidation reactions leading to the formation of volatile compounds responsible for "off-flavors" in the protein of legumes
• the extraction of lipids could constitute an efficient method for producing protein isolates lacking these “off-flavors” and / or whose flavor is more stable during storage, in particular due to the oxidation of the residual lipids.
• the main cause of the development of these “off-flavors” during harvesting, processing and storage is oxidation of unsaturated fatty acids, in particular linoleic and linolenic acids.
• cyclodextrins which are cyclic oligosaccharides composed of several units of glucopyranose (C6H10O5) linked by an a- (1, 4) bond.
• C6H10O5 cyclic oligosaccharides composed of several units of glucopyranose
• ⁇ -cyclodextrins, ⁇ -cyclodextrins and ⁇ -cyclodextrins consisting of 6, 7 and 8 glucopyranoses, respectively.
• b-cyclodextrin has been tested to remove residual lipids and phospholipids from soy protein isolates from delipidated flour.
• a legume protein in particular a legume protein isolate, even more particularly a pea protein isolate having a low lipid content.
• a legume protein isolate the legume being chosen in particular from peas and field beans, preferably pea, characterized in that it contains between 7 g and 9 g, preferably between 7.5 g and 8.5 g of total lipids per 100 g of proteins.
• aqueous solution comprising a mixture of phospholipases and b-cyclodextrins characterized in that the ratio between phospholipase A2 activity and the quantity of b-cyclodextrins, expressed in units of PLA2 activity per g of b-cyclodextrins , is between 10 and 100, preferably between 20 and 80, even more preferably between 25 and 50;
• a legume protein isolate the legume being chosen in particular from peas and field beans, preferably peas, characterized in that it contains between 7 g and 9 g, preferably between 7.5 g and 8.5 g of total fat per 100 g of protein.
• the protein isolate of legumes is a protein isolate of peas.
• protein isolate should be understood in the present application as a composition having a protein content greater than 70%, preferably greater than 80%, even more preferably greater than 85%, this percentage being understood as dry matter of said composition.
• the protein content is calculated using any methodology well known to those skilled in the art. In particular, a determination of the total nitrogen is carried out by Kjeldahl which is multiplied by the coefficient 6.25.
• Said composition therefore comprises proteins, macromolecules formed from one or more polypeptide chains consisting of the chain of amino acid residues linked together by peptide bonds.
• the present invention relates more particularly to globulins (approximately 50-60% of pea proteins). Pea globulins are mainly subdivided into three sub-families: legumines, vicilins and convicilins.
• legume in the present application will be understood the family of dicotyledonous plants of the order Fabales. It is one of the most important families of flowering plants, the third after Orchidaceae and Asteraceae by number of species. It has approximately 765 genera comprising more than 19,500 species.
• Several legumes are important cultivated plants including soybeans, beans, peas, chickpeas, field beans, peanuts, cultivated lentils, cultivated alfalfa, various clovers, broad beans, carob, licorice, lupine.
• pea in the present application includes the varieties of peas belonging to the genus Pisum and more particularly to the species sativum and aestivum. Said mutant varieties are in particular those called “r mutants”, “rb mutants”, “rug 3 mutants”, “rug 4 mutants”, “rug 5 mutants” and “lam mutants” as described in the article by CL HEYDLEY et al. al. entitled “Developing novel pea starches” Proceedings of the Symposium of the Industrial Biochemistry and Biotechnology Group of the Biochemical Society, 1996, pp. 77-87.
• total lipids in the present application is defined as all the lipid molecules without distinction. They thus include triglycerides, phospholipids, free fatty acids.
• the lipids are assayed by acid hydrolysis, followed by extraction with hexane and a specific assay lipids thus extracted, preferably by gas phase chromatography. The preferred method is described below.
• the legume of the protein isolate is pea.
• the protein isolate according to the invention is characterized in that its linoleic acid content is reduced from 20% to 30%, preferably 25%, relative to the content present in the legume seed.
• the protein isolate may have the advantage of not containing traces of organic solvent, that is to say containing less than 100 ppm of solvent relative to the dry mass of isolate.
• the isolate preferably contains less than 10 ppm relative to the dry mass of the isolate, while preferentially does not include any at all.
• organic solvent is meant a solvent made from molecules comprising at least one carbon atom. Rather, the isolate can include inorganic solvents, typically water. This is an advantage over isolates made by a process comprising a step of extracting lipids with an organic solvent such as hexane.
• the protein isolate of the invention can exhibit good functional properties, by good oil or water retention.
• aqueous solution comprising a mixture of phospholipases and b-cyclodextrins characterized in that the ratio between phospholipase A2 activity and the quantity of b-cyclodextrins, expressed in units of PLA2 activity per g of b-cyclodextrins , is between 10 and 100, preferably between 20 and 80, even more preferably between 25 and 50;
• step 1 is carried out by suspending legume proteins characterized in that they are composed of more than 50% of globulins, preferably more than 70%, even more preferably more than 80% .
• globulins can be easily obtained by grinding the seed into flour, then suspending the seed in water and separating the fibers & starch using hydrocyclones and centrifuges. The supernatant solution enriched in proteins will then be rectified to isoelectric pH (approximately 4.5) and undergo controlled heating, in order to separate the globulins in a coagulate which flocculates.
• isoelectric pH approximately 4.5
• water any type of water suitable for the extraction of protein for food consumption.
• potable, carbon-free or demineralized water will be used.
• the ratio between phospholipase A2 activity and the amount of b-cyclodextrins, expressed in units of PLA2 activity per g of b-cyclodextrins is between 10 and 100, preferably between 20 and 80, even more preferably between 25 and 50.
• the amount of ⁇ -cyclodextrins is calculated relative to the amount of total lipids in the protein isolate. This quantity varies between 0.04 and 0.8 g / g of lipids.
• the amount of phospholipase is calculated in order to be in accordance with the ratio described above.
• Phospholipases are enzymes which have the action of hydrolyzing phospholipids.
• a phospholipase which can be used in the context of the process according to the invention is a type A2 phospholipase, in particular the PLA2 Nagase 10P / R produced by Nagase ChemteX Corporation, and which is derived from Streptomyces violaceoruber N B RC 15146.
• the PLA2 activity is measured with soya lecithin as a substrate. This is placed at 37 ° C and pH 8.0, and the activity is measured using, for example, a Wako NEFA-C Test enzyme kit (Wako Pure Chemical Industries, Ltd.). One unit of enzyme activity corresponds to the hydrolysis of 1 pmol of fatty acid in one minute.
• the PLA2 Nagase 10P / R enzyme thus titrated 100,000 U / g.
• Cyclodextrins are cyclic oligosaccharides composed of several units of glucopyranose (CeH-ioOs) linked by an ⁇ - (1, 4) bond. The most common are a-cyclodextrins, b-cyclodextrins and g-cyclodextrins consisting of 6, 7 and 8 glucopyranoses, respectively.
• One of the main advantages of cyclodextrins is their ability to form inclusion complexes with various compounds due to their "conical cylinder" structure.
• the drying of the composition can be done by any method known per se, preferably by lyophilization, passage through a dryer drum or atomization (“spray-drying"), in particular by freeze-drying.
• An advantage of the method is that it may not include an ultrasound treatment step.
• An ultrasound treatment step known per se, consists of a step of applying ultrasound to material (flour, isolate, etc.) during the process.
• the BCD (betacyclodextrin) used is Kleptose® from the company Roquette.
• the phospholipase used is Nagase PLA2 10P / R, diluted to a concentration of 1% w / w with demineralized water.
• the solution also contains 0.5% NaCl and 0.1% CaCl2.
• Example 1 a Production of a protein isolate of lequmineuse according to the invention from globulins previously extracted
• the globulins are extracted according to a conventional extraction. Yellow pea seeds are used in the present example. After shelling the outer fibers on a hammer mill, pea seeds are ground to obtain a flour. This is then soaked in water at the final concentration of 16.5% by weight of dry matter relative to the weight of said suspension, at a pH of 6.5, for 30 minutes at room temperature. The flour suspension at 25% by weight of dry matter is then introduced into a battery of hydrocyclones, separating a light phase consisting of the mixture of proteins, internal fibers (pulps) and soluble and a heavy phase, containing the starch. The light phase at the outlet of the hydrocyclones is then brought to a dry matter content of 11.2% relative to the weight of said suspension.
• the internal fibers are separated by passing them through centrifugal decanters of the WESPHALIA type.
• the light phase at the outlet of the decanter centrifuge contains a mixture of proteins and solubles, while the heavy phase contains the pea fibers.
• the proteins are coagulated at their isoelectric point by adjusting the light phase at the outlet of the decanter centrifuge to a pH of 4.6 and heating this solution at 70 ° C. for 4 min. After coagulation of the proteins, a protein floc is recovered which is mainly composed of globulins.
• the protein floc is resuspended in demineralized water and is then introduced into the reactor where the reagents, bq ⁇ and PLA2 are added, under specific conditions of temperature (45 ° C) and pH (8.5 ).
• the amounts of bq ⁇ and PLA2 are calculated relative to the residual amount of lipids, in a dosage equal to 0.04 g of bq ⁇ per g of lipids and 0.002 g of a 1% solution of phospholipase Nagase PLA2 10P / R per g of lipids. After 180 min of reaction, the solution is heated at 60 ° C for 10 min in order to inhibit PLA2.
• the treated solution is flocculated for 10 min at 60 ° C at pH 4.5 before being centrifuged twice at 8000g for 10 min to remove the bq ⁇ complexes. Finally, the globulins are resuspended in demineralized water and the pH is raised to 7 before lyophilization.
• Example 1 b Production of a protein isolate of lequmineuse from flour
• This example varies from Example 1a in that the injection point of bq ⁇ and phospholipase is located upstream, during the pea flour suspension step.
• pea seeds are ground to obtain a flour. This is then soaked in a reactor, with water at the final concentration of 16.5% by weight of dry matter relative to the weight of said suspension.
• the reactants, bq ⁇ and PLA2 are introduced into the reactor, and the solution thus obtained is placed under specific conditions of temperature (45 ° C.) and pH (8.5).
• the amounts of bq ⁇ and PLA2 are calculated relative to the residual amount of lipids, in a dosage equal to 0.04 g of bq ⁇ per g of lipids and 0.002 g of a 1% solution of phospholipase Nagase PLA2 10P / R per g of lipids. After 180 min of reaction, the solution is heated at 60 ° C for 10 min in order to inhibit PLA2.
• the flour suspension is then introduced into a battery of hydrocyclones, separating a light phase consisting of the mixture of proteins, internal fibers (pulps) and soluble materials and a heavy phase, containing the starch.
• the light phase at the outlet of the hydrocyclones is then brought to a dry matter content of 11.2% relative to the weight of said suspension.
• the internal fibers are separated by passing them through centrifugal decanters of the WESPHALIA type.
• the light phase at the outlet of the decanter centrifuge contains a mixture of proteins and solubles, while the heavy phase contains the pea fibers.
• the proteins are coagulated at their isoelectric point by adjusting the light phase at the outlet of the decanter centrifuge to a pH of 4.6 and heating this solution at 70 ° C. for 4 min. After coagulation of the proteins, a protein floc is recovered which is mainly composed of globulins.
• Example 1 c Production of a protein isolate of lequmineuse according to the invention from globulins previously extracted, with a BCD / Lipase ratio outside the invention
• the aim of this example is to demonstrate the importance of the CD / Iipase ratio
• yellow pea seeds are used. After shelling the outer fibers on a hammer mill, pea seeds are ground to obtain a flour. This is then soaked in water at the final concentration of 16.5% by weight of dry matter relative to the weight of said suspension, at a pH of 6.5, for 30 minutes at room temperature. The flour suspension at 25% by weight of dry matter is then introduced into a battery of hydrocyclones, separating a light phase consisting of the mixture of proteins, internal fibers (pulps) and soluble and a heavy phase, containing the starch. The light phase at the outlet of the hydrocyclones is then brought to a dry matter content of 11.2% relative to the weight of said suspension.
• the internal fibers are separated by passing them through centrifugal decanters of the WESPHALIA type.
• the light phase at the outlet of the decanter centrifuge contains a mixture of proteins and solubles, while the heavy phase contains the pea fibers.
• the proteins are coagulated at their isoelectric point by adjusting the light phase at the outlet of the decanter centrifuge to a pH of 4.6 and heating this solution at 70 ° C. for 4 min. After coagulation of the proteins, a protein floc is recovered which is mainly composed of globulins.
• the protein floc is resuspended in demineralized water and is then introduced into the reactor where the reagents, bq ⁇ and PLA2 are added, under specific conditions of temperature (45 ° C) and pH (8.5 ).
• the amounts of bq ⁇ and PLA2 are calculated relative to the residual amount of lipids, in a dosage respectively equal to 0.71 g of bq ⁇ per g of lipids and 0.002 g of a 1% solution of phospholipase Nagase PLA2 10P / R per g of lipids. After 180 min of reaction, the solution is heated at 60 ° C for 10 min in order to inhibit PLA2.
• the treated solution is flocculated for 10 min at 60 ° C at pH 4.5 before being centrifuged twice at 8000g for 10 min to remove the bq ⁇ complexes. Finally, the globulins are resuspended in demineralized water and the pH is raised to 7 before lyophilization.
• Example 2 Measurement of total lipids in different isolates according to the invention The total lipids are analyzed as well as the content of various fatty acids present.
• the protocol for analyzing total lipids is as follows:
• the protocol for analyzing the linoleic acid content is as follows:
• the protein content is obtained by measuring the nitrogen content of the sample, which will be multiplied by the coefficient 6.25.
• Example 1a makes it possible to obtain a protein isolate having:

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Citations (6)

• 2019
  • 2019-12-23 FR FR1915459A patent/FR3104906B1/fr active Active
• 2020
  • 2020-12-22 CN CN202080086615.4A patent/CN114828641A/zh active Pending
  • 2020-12-22 WO PCT/FR2020/052598 patent/WO2021130446A1/fr not_active Ceased
  • 2020-12-22 JP JP2022538061A patent/JP7787073B2/ja active Active
  • 2020-12-22 AU AU2020415407A patent/AU2020415407A1/en active Pending
  • 2020-12-22 BR BR112022012525A patent/BR112022012525A2/pt unknown
  • 2020-12-22 US US17/757,806 patent/US12520858B2/en active Active
  • 2020-12-22 MX MX2022007838A patent/MX2022007838A/es unknown
  • 2020-12-22 EP EP20851290.5A patent/EP4081044A1/fr active Pending
• 2025
  • 2025-10-21 JP JP2025177236A patent/JP2026010148A/ja active Pending

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