RU2277794C1 - Method for obtaining of structurally modified rape protein product - Google Patents

Abstract

FIELD: preparing of structurally modified protein products.

SUBSTANCE: method involves modifying protein complex by enzymic hydrolysis process, followed by interrupting said process; before carrying out enzymic hydrolysis process, washing and disinfecting seeds with 0.01-0.05-% solution of sorbic acid having water-seed ratio of (3-5):1 for inactivation of surface microflora; performing enzymic hydrolysis process in single stage by germinating seeds for activation of proteolytic enzymes and partly hydrolyzing therewith of protein complex at seed moisture content of 80-150% and at temperature of 10-25 C for 36-60 hours; stopping hydrolysis process by heating germinated seeds at temperature of 85±5 C for 10-30 min; after stopping of hydrolysis process, frying (extracting oil); grinding oil cake and removing seed enclosures. Method allows biologically modified food product to be obtained, said product having improved fat retaining, foaming, fat emulsifying and other functional properties.

EFFECT: increased biological value and improved functional properties of protein product.

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Description

The invention relates to the production of modified protein products and can be used in the food industry.

The expansion and improvement of the assortment of protein-containing products occurs due to the implementation of technologies for the production of skim flour, concentrates, isolates, etc.

For the fullest use of the potential possibilities of the reserve proteins of oilseeds, various biotechnological modifications of the protein complex are carried out, aimed at obtaining easily digestible forms of proteins.

In this regard, enzymatic hydrolysis methods are widely used to obtain and / or modify protein products. Enzymatic hydrolysis deeply affects the structure of a protein molecule, proceeds under mild temperature conditions, leads to a change in the fractional composition of the protein complex and to a mixture of amino acids and peptides.

A known method of producing protein hydrolysates based on a dispersion of lentil proteins by biotechnology. As a dispersion, a 3% dispersion of lentil protein, obtained by extracting the protein from lentil beans in an alkaline medium, was used. The following proteolytic preparations were used for enzymatic hydrolysis: megaterin G-10X, protosubtilin G-10X, and collagenase preparation obtained from hepatopancreas of king crab (E.E. Kurchaeva. Study of the conditions of enzymatic hydrolysis of lentil proteins // Conference Materials "Progressive Technologies and Equipment for Food industry ", Voronezh, 2004, p.110-114). The disadvantage of this method is the need to use an alkaline extractant, which increases the number of technological operations and leads to the complexity of the process.

A known method of producing protein hydrolyzate from rapeseed using endopeptidase (alkalase), an enzyme produced by a strain of Bacillus Licheniformis, and exopeptidase (Flovourzyme - 1000 MG), which is a complex of exoproteases from Aspergillus orysae (Production and characterization of an extesivedessed rapessed protein Javier, Sanchez-Vioque Raul, Clemente Alfonso, Pedrockt Justto, Bautiste Juan, Milan Francisso // J. Amer. Oil Chem. Soc. - 1999.- 76, No. 7, p. 819-823).

The disadvantage of this method is the need to use a series of enzyme preparations in fairly high concentrations to achieve high performance, which leads to a rise in price of the product.

Closest to the claimed method is a method for producing a food protein product using enzymes of microbiological synthesis and animal origin.

The method will include a phased hydrolysis in the presence of a preservative of isolate proteins, chloroform, mixed with water at a concentration of 0.7-0.9 wt.%.

The hydrolysis at the first stage is carried out by the microbiological synthesis enzyme protosubtilin at a concentration of 1.0-1.5% by weight of the isolate at an initial temperature of 70-75 ° C and a pH of 8.8-9.0, followed by a gradual decrease within 1-2 hours to temperature 45-50 ° C, pH 7.5-8.0.

At the second stage, an enzyme of animal origin is used - homogenate of the pancreas of pigs and / or cattle, at a concentration of 1-20% by weight of the isolate, hydrolysis is continued at 45-48 ° C, pH 7.5-8.0, for 1 , 5-2 hours to an amine nitrogen value of 450-500 mg%.

Precipitation of unhydrolyzed mass is carried out at 85-90 ° C, pH 7.3-7.6 for 20-25 minutes, and its separation is carried out with a gradual decrease in temperature to 65-70 ° C by arbitrary sedimentation of solid particles of the hydrolyzate.

Drying of the product clarified by filtration using depth filters is carried out by spraying in a dryer at a temperature of 130-135 ° С at the inlet of the device and 80-85 ° С at its exit (Application for an image. No. 2001134970 RU, IPC ⁷ A 23 J 1/16 "Food protein product and method of its preparation").

The specified method is not suitable for the production of rapeseed protein product, since it does not exclude the possibility of the formation of toxic compounds during the hydrolysis of glycosinolates contained even in the low glucosinolate varieties of Brassica napus of the Kapustny family.

The disadvantages of the prototype include the multi-stage and technological complexity of the process, the use of a series of enzyme preparations, the poor efficiency of animal products for the hydrolysis of plant proteins.

The objective of the invention is to develop a method with the necessary and sufficient number of technological operations to obtain a food enzyme-modified protein rape product of high biological value with improved functional properties.

The technical result of the invention is to obtain a biomodified protein product of high biological value with improved functional properties: fat-retaining, foaming and fat-emulsifying abilities from meal of partially hydrolyzed rapeseed.

In the method involving the modification of the protein complex by enzymatic hydrolysis with its subsequent stop, before carrying out the enzymatic hydrolysis, washing is carried out at a water-seed ratio (3 ÷ 5): 1 and seed disinfection with a solution of 0.01-0.05% sorbenic acid to inactivate surface microflora, enzymatic hydrolysis is carried out in one step by seed germination to activate proteases and partial hydrolysis of the protein complex at a seed moisture of 80-150%, a temperature of 10-25 ° C for 36-60 hours, stops hydrolysis is carried out by heating the germinated seeds at a temperature of 85 ± 5 ° C for 10-30 minutes after stopping the hydrolysis is carried out degreasing (extraction oil), meal grinding and removal of the seed coat.

The process of seed germination is characterized by the activation of hydrolytic enzymes. The use of preliminary germination makes it possible to obtain rapeseed meal with a modified protein complex, which eliminates operations from the technological process to further modify the functional properties of the protein product, increase efficiency and simplify the technological scheme of the process of obtaining a protein product, which reduces the cost of the obtained protein product without reducing its quality.

Considering the strict sequence of actions of individual proteases and the resistance of storage proteins to their action in the initial period of germination, temperature conditions have been developed that promote the activation of specific proteinases 36-48 hours after seed soaking. The hydrolysis of protease A storage proteins begins, which cleave one or two short peptides, after which the whole molecule, due to conformational changes, becomes accessible to the action of other proteases (proteases B and C), which break down the storage proteins into large peptides.

The proposed conditions for the rape seed germination provide hydrolysis of glucosinolates to the formation of isothiocyanates and nitriles, which are involved in the process of protein synthesis during seedling formation until the possible spontaneous formation of 5-vinyl-2-thiooxazolidone in rapeseed meal.

The studied proteolysis conditions, based on the use of proprietary endoproteases of germinating rapeseed seeds, contribute to the improvement of the fat-holding ability of the protein product, its foaming and emulsifying properties.

The time of germination of rape seeds correlates with the degree of hydrolysis of the storage proteins and affects the formation of certain functional properties in the protein product.

The use of proprietary rapeseed enzymes for preliminary limited hydrolysis of seed proteins made it possible to obtain a protein product that does not require further modification of functional properties.

The combination of operations for the isolation and modification of the protein product made it possible to simplify the process.

The separation of the protein portion of the meal from the seed coat on a sieve increased the efficiency of the process and reduced the cost of the resulting rapeseed protein product without loss of quality.

Thus, by introducing a new set of essential features, it is possible to solve the problem posed by the current state of the art.

The inventive method is illustrated by examples.

Example 1. As a raw material selected seeds of winter colza varieties "Zlata". After washing and disinfecting with a 0.05% sorbic acid solution to inactivate the surface microflora (solution to seed ratio 5: 1), the seeds were germinated for 36 hours at a temperature of 25 ° C, at a humidity of 80%, to activate proteases and partially hydrolyze them of the protein complex hydrolysis was stopped by inactivation of enzymes at 85 ± 5 ° С for 10 minutes, then oil was extracted, fat-free meal was crushed, and modified protein flour was enriched by removing seed coatings on a sieve with a mesh diameter of 0.5 mm.

Example 2. The feedstock and the sequence of technological operations, as in example 1.

Preliminary disinfection of seeds was carried out with a 0.03% sorbic acid solution at a ratio of solution - seeds of 4: 1. Germination of prepared seeds was carried out for 48 hours at a temperature of 20 ° C at a humidity of 120%, hydrolysis was stopped at 80 ± 5 ° C for 20 minutes

Example 3. As a raw material selected seeds of winter rape varieties "Otradnensky". The sequence of technological operations, as in example 1.

Preliminary disinfection of seeds was carried out with a 0.01% solution of sorbic acid at a ratio of solution - seeds of 3: 1. Germination of prepared seeds was carried out for 60 hours at a temperature of 10 ° C at a humidity of 150%, hydrolysis was stopped at 80 ± 5 ° C for 30 minutes.

Biochemical parameters of the products obtained by the proposed method are presented in the table.

The determination of biological value was carried out on the test object Tetrachymena pyryphormis relative to the standard protein (casein).

Table Protein product obtained Functional properties,% The relative biological value,% Fat retention capacity Foaming ability Fat emulsifying ability According to the prototype method 121 11.6 76 52 Declared
way Example 1 221 15.7 169 77 Example 2 240 17.1 170 76 Example 3 117 20,0 240 71

As tests have shown, limited hydrolysis by endoproteases by seed germination allows to obtain the target product with high biological value and improved functional properties, without the use of acid and alkaline reagents, technical and animal enzyme preparations.

Claims (1)

A method involving the modification of a protein complex by enzymatic hydrolysis with its subsequent stop, characterized in that before carrying out enzymatic hydrolysis, washing is performed at a ratio of water: seeds (3-5): 1 and seed disinfection with a solution of 0.01-0.05% sorbic acid, enzymatic hydrolysis is carried out in one step by seed germination at a humidity of 80-150%, a temperature of 10-25 ° C for 36-60 hours, the hydrolysis is stopped by heating the germinated seeds at a temperature of (85 ± 5) ° C for 10 -30 min, after about SETTING hydrolysis is carried out degreasing oil extraction, grinding the meal and removing the seed coats.