RU2604197C1 - Method of producing protein hydrolysate from grain raw material - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: invention relates to food industry. Triticale grains are milled and mixed with water at water duty 1:4 to produce grain suspension. Enzyme preparations of proteolytic action are added to grain suspension. Hydrolysis is carried out in two stages at temperature of 50 °C and pH 5.0-6.0, duration of each hydrolysis stage is 2 hours. First state comprises adding enzyme preparations of bacterial origin Neutrase 1.5 MG and Alcalase 2.4 L FG in amount of 0.5-0.75 units PS/g of milled raw material, and at second stage - mushroom origin Protease GC-106 in amount of 0.75-1.0 units PS/g of milled raw material. Enzyme preparations are inactivated by heating at temperature of 80 °C for 7-10 minutes. Solid phase is removed and clarified hydrolysate is obtained.

EFFECT: invention increases output of product, simplifies process of production and reduces its duration, and enables to manufacture a product with defined functional properties, specifically gluten-free, hypoallergenic, easily digestible.

1 cl, 1 tbl

Description

The invention relates to the food industry, namely to obtain a protein hydrolyzate from grain raw materials, and can be used in the manufacture of products with reduced allergenicity.

Enzymatic modification of plant-derived proteins, including cereal proteins, represents an important stage in promising technologies for the deep processing of grain raw materials. As a result of the modification of the protein components of grain and flour with the use of enzymatic preparations of proteolytic action and compositions based on them, hydrolysis products with a specific peptide profile and a set of amino acids with specific properties can be obtained.

In this regard, methods are widely used to obtain and / or modify protein products. Enzymatic hydrolysis using various proteolytic enzyme preparations deeply affects the structure of the 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.

For example, a method is known for producing a protein hydrolyzate from rapeseed using endopeptidase (Alkalase 2.4 L FG), an enzyme produced by the Bacillus licheniformis strain and exopeptidase (Flovourzyme-1000 MG), which is a complex of exoproteases from Aspergillus orysae (Production and characterization of an extesive rapessed protein haydrolysate / Vioque Javier, Sanchez-Vioque Raul, Clemente Alfonso, Pedrockt Justto, Bautiste Juan, Milan Francisso // J. Amer. Oil Chem. Soc. - 1999. - V. 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.

For the production of protein concentrates and hydrolysates, various grain raw materials are widely used. One of the promising types of such raw materials is triticale grain. It is known that the triticale protein is characterized by a well-balanced amino acid composition. Like wheat and rye proteins, the first limiting amino acid of the triticale protein is lysine, the second is threonine. At the same time, the lysine content in triticale is higher compared to wheat, therefore, the protein of this culture is biologically more complete than wheat protein, as a result of which the use of triticale grains in food production can significantly increase their biological value (Mironenko N.N. Peculiarities of the amino acid composition of triticale grain protein / Non-traditional natural resources, innovative technologies and products. - 2002. - Issue 6. - P. 157-164).

Therefore, the search for new methods for producing protein hydrolysates from grain raw materials using various enzyme preparations is promising.

A known method of producing protein hydrolyzate from rice flour with sequential use of amylolytic, xylanase and proteolytic enzyme preparations, followed by separation from soluble substances (RU 2012147113 A, 05.20.2014).

The disadvantage of this method is the undesirable additional use of alkali and acid.

A known method of producing a high-protein base from wheat grain (RU2453126 C1, 06/20/2012) for preparing a food product, including grinding wheat grain to a flour grinding fraction, moistening it with water in a ratio of 1: 2, enzymatic hydrolysis of Protosubtilin G ZX at a concentration of 0.3% for 22 hours at 45-50 ° C, pH 6.5-7.5, separation of the wetted mass by centrifugation into sediment and centrate.

The disadvantage of this method is the length of the process.

The closest analogue of the claimed invention is a method for preparing a grain concentrate, which involves cleaning seeds of cereal grains from impurities, soaking, drying, grinding; as the seeds of cereal grains, triticale grain is used, which is soaked in a buffer solution consisting of citric acid and sodium citrate with a pH of 4.5, containing the enzyme preparation Celloviridin G20x or a complex enzyme preparation based on phytase F-4.2 V FD-UF in an amount of 0 , 05-0.1% of the mass of dry solids of the grain with a grain: buffer ratio of 2: 1 and a temperature of 40-50 ° C for 9-12 hours until the grain reaches a moisture content of 40-43%, preventing grain germination, grain grinding is carried out after drying to a moisture content of 11-14% and a particle size of 0.05-0.08 mm (RU 2344609 C1, 01/27/2009).

The disadvantage of this method is that with prolonged soaking of the grain, amylolytic and proteolytic enzymes are activated, which hydrolyse starch and gluten proteins, which leads to a decrease in the quality of the finished product (bread and bakery products) in which the obtained concentrate is used.

The technical result of the present invention is to increase the yield of protein from raw materials, carry out deeper hydrolysis, expand the assortment of enzyme preparations used in the technological process, shorten the duration of the technological process and simplify (exclude the long stage of soaking) and expand the scope of use of the finished product.

The claimed technical result is achieved by the fact that the method of producing a protein hydrolyzate from grain raw materials involves grinding grain raw materials, mixing the crushed grain raw materials with water with a 1: 4 hydraulic module to obtain a grain suspension, introducing proteolytic enzyme preparations into the grain suspension, carrying out hydrolysis in two stages, moreover, at the first stage, enzyme preparations of bacterial origin are introduced, and at the second stage, fungal origin, inactivation of enzyme preparations by heating At a temperature of 80 ° C for 7-10 minutes, separation into a hydrolyzate and a solid residue, while triticale grain is used as a raw material of grain, Neutrazu 1.5 MG and Alcalase 2.4 L FG in the amount of enzyme preparations of bacterial origin 0.5-0.75 units each PS / g of crushed raw materials of mushroom origin - Protease GC-106 in the amount of 0.75-1.0 units. PS / g of crushed raw materials, hydrolysis is carried out at a temperature of 50 ° C and a pH of 5.0-6.0, the duration of each stage is 2 hours.

The choice of quantities of these enzyme preparations is due to the need to conduct an effective hydrolysis process. It was found that in this limit of the concentration of enzyme preparations a high reaction rate of hydrolytic cleavage of the substrate is observed, and the quality of the finished product does not deteriorate. With an increase in the dosage of proteolytic enzyme preparations above the declared, there is an unreasonable rise in the price of the obtained product, while a decrease below the declared, there is no deep cleavage of protein substances of triticale grain.

The hydromodule of enzymatic hydrolysis is selected based on the characteristics of the feedstock. At a lower value of the hydromodule, thick viscous media are obtained, as a result of which the process of sorption of enzymes and removal of reaction products is hindered. With a hydraulic module above 1: 4, there is no increase in the yield of protein hydrolyzate.

The method is as follows.

The crushed triticale grain, characterized by a 100% pass through a sieve d = 1 mm, is mixed with water in a ratio of 1: 4. A buffer (phosphate-citrate buffer pH 5.5) is added to regulate a given pH level. Next, proteolytic enzyme preparations of bacterial origin, Neutraz 1.5 MG and Alcalase 2.4 L FG, in an amount of 0.5-0.75 units each, are added to the obtained grain suspension. PS / g of crushed raw materials, hydrolysis is carried out at a temperature of 50 ° C for 2 hours, then a proteolytic enzyme preparation of fungal origin, Protease GC-106, is added in an amount of 0.75-1.0 units. PS / g of crushed raw materials and carry out hydrolysis at a temperature of 50 ° C for 2 hours. Next, inactivation of enzyme preparations is carried out by heating at a temperature of 80 ° C for 7-10 minutes. The resulting suspension is separated in a centrifuge at 5000-6000 rpm for 15 minutes to obtain a clarified hydrolyzate and a solid residue. The obtained hydrolyzate can additionally be dried by any method known in the art (spray drying, freeze drying, etc.). The protein yield is 80-85% of its initial content in the crushed raw materials, depending on the amount of enzyme preparations introduced.

In addition, a chromatographic analysis of the obtained hydrolyzate was carried out using the above enzyme preparations in an amount of: Neutraz 1.5 MG - 0.5 units. PS / g of raw materials, Alcalase 2.4 L FG - 0.5 units. PS / g of raw materials, Protease GC-106 - 0.75 units. PS / g of raw materials. For this, 5 ml of the supernatant was applied to a column filled with Sephadex G-75 gel. Elution was performed with distilled water. The volume of collected fractions is 4 ml. The optical density of the eluate in the fractions was recorded at a wavelength of 280 nm. As a control, an aqueous extract of triticale flour was used, obtained at a ratio of crushed raw materials: water 1:20. Table 1 presents data on the fractional composition of the obtained hydrolyzate.

The analysis of the data presented indicates that the use of MEK, which includes proteolytic enzymes with different specificity of action, fundamentally changed the picture of protein elution and allowed almost completely hydrolyzing the proteins, including, possibly, high molecular weight products of gluten protein proteolysis, which partially turn into solution. So, the high molecular weight fraction with a molecular weight of more than 70,000 and the fraction with a molecular weight of 40,000 ÷ 30,000 Yes are practically absent, while the amount of low molecular weight peptides and amino acids in the hydrolyzate increased by about 2.5-3.0 times compared to the control. This fact is probably due to the fact that the effect of the enzyme preparations used is directed exclusively to soluble proteins, and they do not contribute to the transition of proteins from an insoluble form to a soluble one.

In this connection, the obtained data allow us to position the hydrolyzate obtained using the composition based on enzyme preparations of proteases as a possible component of products with functional properties, namely, as a component of preventive products with reduced allergenicity, since amino acids and low molecular weight peptides, unlike proteins and high molecular weight ones peptides, do not have species specificity and do not cause the production of antigens (immune response); as well as a possible component of gluten-free flour and other foods for the elderly and people with gluten intolerance (celiac disease).

Claims (2)

1. A method of producing a protein hydrolyzate from grain raw materials, including grinding grain raw materials, mixing the crushed grain raw materials with water with a 1: 4 hydraulic module to obtain a grain suspension, introducing proteolytic enzyme preparations into the grain suspension, carrying out hydrolysis in two stages, and in the first stage enzymatic preparations of bacterial origin are introduced, and on the second - fungal origin, inactivation of enzyme preparations by heating at a temperature of 80 ° C for 7-10 minutes, removing solid phase and obtaining a clarified hydrolyzate, while triticale grain is used as a raw material of grain, Neutraz 1.5 MG and Alcalase 2.4 L FG in quantities of 0.5-0.75 units are used as bacterial enzyme preparations. PS / g of crushed raw materials of mushroom origin - Protease GC-106 in the amount of 0.75-1.0 units. PS / g of crushed raw materials, hydrolysis is carried out at a temperature of 50 ° C and pH 5.0-6.0, the duration of each stage of hydrolysis is 2 hours 2. The method according to claim 1, further comprising drying the hydrolyzate to obtain a dry modified protein product.