SU1734646A1 - Method for production of protein additive - Google Patents

Abstract

Use: food industry and feed production, namely the development of microbiological tests to determine the nutritional value of plant materials. SUBSTANCE: method involves partial hydrolysis of vegetable raw materials, which is carried out in a 0.3-0.5 n alcoholic alkali solution, and subsequent purification by extraction with solvents, in particular, hexane and chloroform, which allows selectively extracting more balanced amino acid composition peptides. 4 ex. 2 tab.

Description

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The invention relates to the food industry and feed production, in particular to methods for producing nutritional protein supplements.

Purified and / or modified proteins, obtained from crushed plant raw materials, in particular from plant seeds, have improved properties and are suitable for use in the food industry as special additives (1-4).

Methods are known for modifying a protein to increase its nutritional value, which are as follows: acetylate and succinate the protein and use purified bifunctional protein derivatives (1); protein is treated with succinic anhydride and acetic acid and purified by gel chromatography (2); by hydrolysis in the medium Ca (OH) 2 + NaOH for 2 days at 65-95 ° С in water vapor in the pipeline with

subsequent drying (3). The significant drawbacks of the known methods include multistage, duration, use of toxic reagents. In method (3), there is no purification from lipid, carbohydrate impurities, which form, during such a long hydrolysis, resinous substances that reduce the nutritional value of the resulting mixture of amino acids.

The closest in technical essence and the achieved effect is a method of obtaining a feed additive from corn extract - waste starch and glucose production, including the hydrolysis of a partially evaporated protein extract by enzymes or acids (6-7 h) before splitting 60-70% of the contained in it protein on amino acids with the subsequent neutralization and drying (4)

To perform the work provided for in this method, proteoli are necessary.

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tic enzymes or 6-7 hour hydrolysis of 5-5.5% H2S04. In addition, spraying at a coolant temperature of 280-310 ° C to a solids content of 92-94% is used to dry the hydrolysates. In this method, there is also no purification step of lipid and carbohydrate components. As mentioned above, the absence of this operation reduces the nutritional value of the supplement. In addition, the hydrolyzate is contaminated with salts introduced during hydrolysis and neutralization, which are harmful components in the diet.

The aim of the invention is to simplify, speed up the method of modification and increase the nutritional value of the protein supplement.

This goal is achieved by a method involving protein hydrolysis in a 0.3 N solution of alcoholic alkali for 1 hour, followed by extraction with hexane, 2-fold dilution with water, acidification, and extraction with chloroform. The mild hydrolysis conditions leading to a slight disruption of peptide bonds allow the purification method to be applied with hexane and chloroform. In addition, unwashed substances are transferred to the hexane fraction, and fatty acids to the chloroform fraction. At the final stage of chloroform extraction, the upper aqueous-alcoholic layer containing inorganic salts (Na2SO4) is also removed. The protein mixture, which is at the interphase of chloroform - aqueous ethanol, is removed from the separatory funnel and dried in air;

The resulting preparation has a higher (1.3-1.6 times) nutritional value, determined by a microbiological method, compared to unmodified zein. The increased nutritional value of the drug is due to partial hydrolysis of the protein and, probably, the fact that selective extraction of peptides with more amino acid composition occurs. It is the use of partial hydrolysis followed by solvent extraction that provides the proposed method for the modification and purification of zein.

Example 1.1 g of zein extracted from corn grain flour using conventional methods is poured with 25 ml of 0.3 N solution of NaOH in alcohol, boiled for 1 hour under reflux, cooled in a separatory funnel, diluted twice with water, shaken three times with 20 ml of hexane. The hexane fraction is discarded. 6N H2504 is acidified to pH 2-3, shaken twice with chloroform, the lower layer is removed. A layer of solvents

containing protein in suspension, placed in a beaker and air dried.

Get 0.45-0.50 g of the drug modified zein with high nutritional value, determined by microbiological method for the increase in biomass you. subtilis 72 (Table 1).

From the example it can be seen that partial hydrolysis and purification by solvent extraction results in a protein with an increased nutritional value.

Example 2. 1g of Zein is poured with 25 ml of 0.1; 0.2; 0.3; 0.4; 0.5; 0.6; 0.7 N NaOH solution in alcohol, boil for 1 h and proceed further in the same way as in Example 1. Increased alkali concentration of more than 0.5 n leads to under-treatment (yellow color of the preparation), reducing the nutritional value of the product (Table 1). 2) Thus, the optimum concentration of alcoholic alkali NaOH to obtain a good yield of purified nutritional protein is in the range of 0.3-0.5 n.

Example 1 g of zein is poured over 25 ml of 0.3 n alcoholic NaOH and boiled for 50 minutes, 1 hour, 1 hour, 10 minutes, 1 hour, 30 minutes, 1 hour and 50 minutes, 2 hours. Proceed as follows, as in Example 1, the yield is determined product and its nutritional value (Table 2). An increase in the hydrolyzate time of more than 1.5 hours leads to a loss of product yield, and a decrease in the hydrolysis time leads to a decrease in the nutritional value of the drug (less disruption of polypeptide bonds, worse separation of fatty acids, under treatment) (Table 2).

Example 4. 1g of grains is poured with 25 ml of 0.3 N alcoholic NaOH and boiled for 1 hour, 1 hour and 30 minutes, cooled and dried. Get the sediment weighing 1.2 g, which has negligible nutritional value. Thus, the presence of lipid, carbohydrate components and salts, due to the absence of a purification stage, reduces the nutritional value of the drug (No. 13, 14, Table 2).

Thus, the introduction of an additional operation for the purification of zein with solvents made it possible to increase the nutritional value of zein and eliminate the effect of lipid and carbohydrate components and salts.

Examples 1-4 and data table. 2 show that the optimal conditions for the modification of zein are high yield and high nutritional value: the concentration of alcohol alkali is 0.3-0.5 n and the hydrolysis time is 1-1.5 h. Further, it is impractical to increase the hydrolysis time and alkali concentration from - due to a decrease in the product yield, and a decrease in these parameters below 0.3 n and 1 h leads to a deterioration in the degree of purification and, consequently, a decrease in

nutritional value of protein. Thus, the claimed limits (hydrolysis time of 1 h - 1.5 h and alkali concentration 0.3-0.5 n) lead to the maximum positive result.

The use of the present invention in comparison with the prototype will speed up and simplify the process and obtain a protein with enhanced nutritional value.

Claims (1)

Invention Formula The accumulation of biomass you. subtilis 72 in media with normal and modified zein The yield and nutritional value of the modified zein under various modification conditions A method of producing a protein supplement, including hydrolysis of vegetable raw materials, characterized in that, in order to increase the nutritional value of the finished product, protein hydrolysis is carried out in a 0.3-0.5 n alcoholic alkali solution, and the resulting hydrolyzate is treated with hexane and chloroform. Table 1 table 2