RU2800371C1 - Method of obtaining protein hydrolyzate from corn extract (concessed) - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: invention relates to the methods of obtaining protein hydrolysates used as a source of nitrogen in the composition of microbiological nutrient media. The method for obtaining a protein hydrolyzate involves pre-treatment of raw materials. Distilled water is poured into corn extract at the rate of 5 liters per 4 kg of raw material, the mass is boiled for 5 minutes, thoroughly mixed and the mixture is left for 12 hours at room temperature, then distilled water is added until the ratio of raw materials and water is 1:4.5, obtained the solution is heated to a temperature of 80–90°C and pH is set to 7.0–7.4, then the mixture is heated to boiling and at a temperature of 100°C, it is kept for 5 minutes, the mixture is cooled before hydrolysis. The hydrolysis process is carried out with the introduction of a hydrolyzing agent of the pancreas of cattle at a concentration of 25 g/l for 70–72 hours at a temperature of 47–49°C and concentration of hydrogen ions 7.8–8.0 pH units.

EFFECT: invention allows to reduce the time of the technological process of obtaining a hydrolyzate without worsening its physicochemical characteristics.

1 cl, 2 tbl, 7 ex

Description

Application area.

The invention relates to the field of biotechnology, in particular to methods for obtaining protein hydrolysates used as a source of nitrogen in the composition of microbiological nutrient media, and can be used in microbiology, as well as other areas in which nutrient media are designed.

The level of technology.

The attention of specialists in the field of biotechnology, in the development of nutrient bases and media, is increasingly drawn to hydrolysates from plant materials and waste products from their processing. The raw material we offer for obtaining protein hydrolyzate is a by-product of starch and syrup production - corn extract (condensed). The composition of the extract indicates its high nutritional value. It contains about 22.5% protein, 17.0-17.2% carbohydrates, 0.0028% pectin, up to 17% lactic acid. (Condensed corn extract. Irina Bondar. 03/02/2021. http://karotinka.dp.ua/stodentam/ekstrakt-kukuruznyj-sgushhennyj/)

In the prior art, methods are known for producing hydrolysates from corn extract (condensed) (CE).

A method is known for producing an enzymatic hydrolyzate of CE, in which by mixing the hydrolysis components - raw materials and water in a ratio of 1: 1.5, under the action of a hydrolyzing agent (pancreas of cattle (cattle) 30 g / l), the process is carried out at a temperature of 46 ° C, the pH of the mixture is 8.2 for 10 days. The obtained hydrolyzate contains 700 mg% of amino nitrogen. The amount of total nitrogen and the degree of protein digestion are not indicated. (Dissertation for the degree of candidate of biological sciences., Saratov 1994, Gyulushanyan K.S.)

The disadvantage of this method is the duration of the process, the low yield of the finished product (due to the ratio of raw materials and water 1:1.5), with the concentration of amine nitrogen - 700 mg%.

The closest to the declared in terms of the totality of features and the achieved effect is a method for obtaining an enzymatic hydrolyzate, which involves the hydrolysis of CE (the ratio of raw materials to water is 1:6), cattle pancreas 30 g/l is used as a hydrolyzing agent, the process is carried out at a temperature of 46 ° C, a mixture pH of 8.2 for 7-10 days. The result is a product with an amino nitrogen content of 1120 mg%. The amount of total nitrogen and the degree of protein digestion are not indicated. (Patent RU No. 262568. Published on July 28, 2017. Bull. No. 22.). Taken as a prototype.

The reasons preventing the achievement of the technical result indicated below when using the known method, taken as a prototype, include the fact that the raw materials used in it are immediately subjected to hydrolysis, without pre-treatment, thereby incomplete extraction of nutrients into the final product occurs. In addition, the component composition is not optimized, as a result of which the hydrolysis proceeds for a long time.

The essence of the invention

1. The objective of the invention is to reduce the duration of CE hydrolysis and ensure the production of a hydrolyzate with desired properties (the degree of protein cleavage is more than 50%, the content of amine nitrogen is not less than 1120 mg %).

2. Thus, the technical result obtained from the use of this invention is to increase the productivity of the method both by increasing the depth of processing of raw materials, and by reducing the hydrolysis time and ultimately obtaining a product with desired properties.

The specified technical result, in the implementation of the invention, is achieved due to the fact that in the proposed method, before hydrolysis, the raw materials are pre-treated, the process of enzymatic cleavage of the protein is carried out at a temperature of 47-49 ° C and a hydrogen ion concentration of 7.8-8.0 pH units in the presence of cattle pancreas in the amount of 25 g/l for 3 days (70-72 hours).

The ratio of raw materials and water is essential. So, when diluting raw materials with less water, it is impossible to achieve the required degree of protein cleavage due to lack of water. An excess amount of water is also not advisable, since the duration of the hydrolysis process increases. The degree of hydrolysis is also affected by the bioavailability of the substrate, which in this case is determined by the pretreatment of the raw material.

Depending on the processing of CE at the stated substrate/enzyme ratio, it is possible to obtain the target product that meets the requirements for the quality of similar preparations in terms of physicochemical characteristics while reducing the duration of the process.

The presence of a causal relationship between the set of essential features of the claimed object and the achieved technical result is shown in table 1.

The proposed set of features allows you to reduce the duration of the hydrolysis process to 3 days (70-72 hours) instead of 7-10 days according to the prototype method, the selected ratio of hydrolysis components allows you to get the final product with the desired properties (the degree of protein cleavage is more than 50%, the content of amine nitrogen is not less than 1120 mg%).

The analysis of the prior art made it possible to establish that no source was found that is characterized by features identical to all the essential features of the claimed invention. Therefore, the claimed invention meets the condition of "novelty".

An additional search for known solutions has shown that the claimed invention does not follow explicitly from the prior art for a specialist, therefore, the claimed invention meets the "inventive step" condition.

Information confirming the possibility of carrying out the invention to obtain the above technical result.

The method of preparation of the protein hydrolyzate is as follows.

CE processing.

Distilled water is poured into a stainless steel container at the rate of 5 liters per 4 kg of corn extract. Then the whole mass is boiled for 5 minutes for better extraction of nutrients. After boiling, the resulting CE solution was thoroughly mixed and the mixture was left for 12 h at room temperature. Then, 13 liters of distilled water are added to the resulting CE solution. The resulting solution is heated to a temperature of 80-90°C and the concentration of hydrogen ions is set with a 20% sodium hydroxide solution of 7.0-7.4 pH units.

Characteristics of the solution treated with CE, in terms of physicochemical parameters, are presented in Table 2.

Hydrolysis of treated CE.

Hydrolysis is carried out as follows. The treated CE solution is poured into the reactor through the loading hatch. Next, the mixture is heated to boiling and at a temperature of 100°C incubated for 5 minutes. Then the reaction mixture is cooled to a temperature of 47-49°C and the pH of the mixture is adjusted to 7.8-8.0 with a 40% NaOH solution. In the resulting reaction mixture with a working stirrer, a previously prepared sample of minced meat of the pancreas of cattle is introduced at the rate of 25 g/l. Hydrolysis is carried out with periodic stirring: the first day for 15 minutes every 2 hours, then twice a day for 20 minutes. During the hydrolysis, samples are taken to determine the content of amine nitrogen and the pH of the reaction mixture. The concentration of hydrogen ions is maintained in the range from 7.8 to 8.0 pH units. Hydrolysis is carried out for 3 days (70-72 hours), the end of hydrolysis is determined by the cessation of the increase in the content of amine nitrogen. At the end of hydrolysis, the mixture is boiled for better sedimentation of ballast substances. The resulting hydrolyzate is purified by filtration through belting and evaluated by physico-chemical parameters.

The invention is illustrated by the following examples, but is not limited to them.

Example 1. Hydrolysis of corn extract (condensed) with the introduction of a hydrolyzing agent (bovine pancreas) in an amount of 25 g/l was carried out at a ratio of raw materials and water of 1:3, respectively, the process was carried out at a temperature of 47-49 ° C and a concentration of hydrogen ions of 7.8-8.0 pH units. With this ratio of raw materials and the conditions of the hydrolysis process, the degree of protein cleavage was 58%, the concentration of amine nitrogen was 791.8 mg%, and the duration of the process was 3 days.

Example 2. Hydrolysis of corn extract (condensed) with the introduction of a hydrolyzing agent (bovine pancreas) in an amount of 25 g/l was carried out at a ratio of raw materials and water of 1:4.5, respectively, the process was carried out at a temperature of 47-49 ° C and a concentration of hydrogen ions of 7.8-8.0 pH units. With this ratio of raw materials and the conditions of the hydrolysis process, the degree of protein cleavage was 67%, the concentration of amine nitrogen was 1196 mg%, the duration of the process was 3 days (70-72 hours).

Example 3. Hydrolysis of corn extract (condensed) with the introduction of a hydrolyzing agent (bovine pancreas) in an amount of 25 g/l was carried out at a ratio of raw materials and water of 1:5, respectively, the process was carried out at a temperature of 47-49 ° C and a concentration of hydrogen ions of 7.8-8.0 pH units. With this ratio of raw materials and conditions of the hydrolysis process, the degree of protein cleavage was 52%, the concentration of amine nitrogen was 646 mg%, and the duration of the process was 3 days.

Example 4. Hydrolysis of corn extract (condensed) with the introduction of a hydrolyzing agent (bovine pancreas) in an amount of 25 g/l was carried out at a ratio of raw materials and water of 1:4.5, respectively, the process was carried out at a temperature of 40-46 ° C and a concentration of hydrogen ions of 7.4-7.7 pH units. With this ratio of raw materials and the conditions of the hydrolysis process, the degree of protein cleavage was 58%, the concentration of amine nitrogen was 810 mg%, and the duration of the process was 6 days.

Example 5. Hydrolysis of corn extract (condensed) with the introduction of a hydrolyzing agent (bovine pancreas) in an amount of 25 g/l was carried out at a ratio of raw materials and water of 1:4.5, respectively, the process was carried out at a temperature of 49-51 ° C and a concentration of hydrogen ions of 7.4-7.7 pH units. With this ratio of raw materials and the conditions of the hydrolysis process, the degree of protein cleavage was 55%, the concentration of amine nitrogen was 788.7 mg%, and the duration of the process was 4 days.

Example 6. Hydrolysis of corn extract (condensed) with the introduction of a hydrolyzing agent (bovine pancreas) in an amount of 25 g/l was carried out at a ratio of raw materials and water of 1:4.5, respectively, the process was carried out at a temperature of 49-51 ° C, a concentration of hydrogen ions of 8.1-8.2 pH units. With this ratio of raw materials and the conditions of the hydrolysis process, the degree of protein cleavage was 52%, the concentration of amine nitrogen was 890.3 mg%, the duration of the process was 3 days.

Example 7. Hydrolysis of corn extract (condensed) with the introduction of a hydrolyzing agent (bovine pancreas) in an amount of 25 g/l was carried out at a ratio of raw materials and water of 1:4.5, respectively, the process was carried out at a temperature of 40-46°C and a concentration of hydrogen ions of 8.1-8.2 pH units. With this ratio of raw materials and the conditions of the hydrolysis process, the degree of protein cleavage was 51%, the concentration of amine nitrogen was 910.5 mg%, the duration of the process was 5 days.

Thus, the claimed method for preparing a protein hydrolyzate from corn extract (condensed) (example 2) provides a hydrolyzate with a high degree of protein cleavage and a given amine nitrogen content, and therefore can be used in the design of nutrient media for cultivating microorganisms.

EFFECT: invention allows to reduce the time of the technological process of obtaining a hydrolyzate without worsening its physicochemical characteristics.

Claims (1)

A method for obtaining a protein hydrolyzate, which involves the hydrolysis of corn extract condensed with the introduction of a hydrolyzing agent of the pancreas of cattle, characterized in that pre-treatment of the raw material is carried out, in which distilled water and corn extract are poured at the rate of 5 l per 4 kg of raw material, the mass is boiled for 5 minutes, thoroughly mixed and the mixture is left for 12 hours at room temperature, then distilled water is added until the ratio of raw materials and water is 1:4.5 , the resulting solution is heated to a temperature of 80-90°C and the pH is set to 7.0-7.4, then the mixture is heated to boiling and kept at a temperature of 100°C for 5 minutes, the mixture is cooled before hydrolysis, the process is carried out for 70-72 hours at a temperature of 47-49°C and a hydrogen ion concentration of 7.8-8.0 units. pH in the presence of bovine pancreas at a concentration of 25 g/l.