SU1074901A1 - Process for preparing proteinaceous hydrolyzate from collagenaceous wastes of production of protein sausage skin - Google Patents

Abstract

A method for producing protein hydrolyzate from collagen-containing waste production of protein sausage shell, including hydrolysis of the feedstock, characterized in that, in order to obtain a hydroplating suitable for long. preparing microbiological nutrient media to control sterility; the hydrolysis of the raw material is carried out in a 0.4-0.3% hydrochloric acid solution for 30-40 minutes at. 100-120 s, and then subjected to enzymatic hydrolysis, which is carried out with pancreatic gland, at 40-42 s for 2-3 days. (L 4; ii with

Description

The invention relates to the microbial industry of logic, and specifically to methods for preparing bases for nutrient media. A known method for producing a nutrient base by enzymatically hydrolyzing casein SP. The disadvantages of the method are the multitude of processes, a small output of the hydrolyzate from 1 kg of raw material to its scarcity and high cost. In addition, casein is a food base, produced by the dairy industry. In this regard, its use for the preparation of nutrient media is irrational. There is also known a method for producing protein hydrolyzate from collagen-containing waste from the production of cobalt protein, including hydrolysis of the feedstock. The hydrolysis of the original collagen-containing raw material is carried out with lime milk (calcium hydroxide), followed by neutralization and purification of calcium salts C) 6 by treatment of the hydrolyzate with phosphoric acid, and then with orthophosphoric disubstituted alkali metal salt or ammonium salt. Collagen-containing raw material, which is a waste product of the production of protein sausage casing, is taken as a source material. After hydrolysis, the reaction mixture is subjected to a hydrothermal treatment for 10–16 hours at BO-CU and atmospheric pressure. After the end of the hydrothermal treatment, the product is subjected to mechanical cleaning of the sludge by centrifuging or filtering. In this case, the sludge is a precipitate of calcium hydroxide and partially mechanical impurities. Next, a chemical purification of the product is carried out, which consists in treating the hydrolysis product with phosphoric acid at pH 7-8. In this case, the main precipitation of the calcium phosphate disubstituted salt occurs. The remaining calcium in the solution at these values is precipitated by adding sodium phosphate (ortho) disubstituted 12-aqueous sodium, stirred for 1 hour, and kept in a sump for 24-36 hours. The precipitated calcium phosphate (ortho) calcium salt is separated by centrifugation and filtration. The resulting protein hydrolyzate is concentrated in an evaporator to 40-60% or dried to a powder in a dryer. Instead of the phosphoric acid (ortho) salt of an alkali metal, the phosphoric acid (ortho) salts of other alkali metals or the ammonium salt of C2j are used. The disadvantages of the method are the duration of the process, multistage, the use of scarce, limctated reagents (phosphoric acid). Consequently, the production of this drug is expensive. The resulting preparation is used mainly in cosme; 7, it is chemical, as a basis for nutrient media it cannot be used immediately, since according to the available materials for this raw material, it has a small degree of cleavage equal to 11-17%. The aim of the invention is to obtain a hydrolyzate suitable for the preparation of microbiological nutrient media for controlling sterility. This goal is achieved by the fact that according to the method of obtaining protein hydrolyzate from collagen-containing waste production of protein sausage casing, including hydrolysis of the raw material, the latter is carried out in about 4-0., 3% hydrochloric acid solution for 30-40 minutes at 100-120 ° C, and then the raw material is subjected to enzymatic hydrolysis, which is carried out with the help of: I put it on the pelvic gland at 40-42 ° C for 2-3 days. At a lower concentration of hydrochloric acid, i.e. 0.1-0.2%, hydrolysis of only a non-tanned shell of a small degree of proliferation occurs. A sheath with a normal degree of tanning remains in the hydrolyzate as a viscous sediment. At an acid concentration of 0.5% and above, hydrolysis and dubbing tanning and non-tanned shells occur simultaneously, but the hydrolyzate occurs. in this case, it has a significant ash content (in the form of mineral salts) and an overestimate of chlorine ions, which is undesirable to be based on nutrient media. The concentration of the acid is 0.3-0.4% gives the sloughing and hydrolysis simultaneously of all types of shell waste with a permissible content of chlorine ions, in the hydrolyzate and a reduced ash content, which improves the quality of the hydrolyzate as a base. Saline acid in concentrates, 3–0.4%. At 120–160 ° C for 0–30 min, protein waste is destructed by 11–15%, as a result of which products are D) (1drolysis contains a mixture of mineral acids and peptides that are not subjected destruction, the minimum amount of free acid, which have good physical properties, then allows the hydrolyzate to be used as the basis for nutrient media, followed by enzymatic hydrolysis.

Example 1. Baseline data: hydrochloric acid concentration 0.3%. The duration of hydrolysis is 3 days 40 minutes, the temperature of hydrolysis is 120s

300 kg of ground sausage casing wastes are loaded into the reactor, 435 l of water and 3.75 of concentrated hydrochloric acid are added, the hydrolysis of the raw material is carried out for 40 minutes at 120 ° C in a 0.3% hydrochloric acid mixture. The liquid hydrolyzate is dried on a single-shaft atmospheric drier.

The obtained dry product is subjected to enzymatic hydrolysis using pancreatic minced meat and tap water (at the rate of 1 kg of dry powder, 1 kg of minced pancreatic meat per .10 l of water), alkalinized with 20% KxsOK to pH 7.8, preserve once, added 1.0% chloroform. Hydrolysis is carried out for 3 days at.

Chemical indicators of the obtained base:

The duration of the process, day3

Total nitrogen, mg% 1300 Amine nitrogen, mg% 864.36 Chlorides,% 0.8SO

TryptophanSledges

Peptone,% 1.4

Dry residue,% 10.7 This example confirms the obtaining of a nutritional basis in the shortest possible time with the necessary degree of cleavage and physico-chemical indicators.

With a decrease in hydrolysis time of less than 30 minutes, a decrease in perurature tag of less than 100 ° C, but with the same concentration of hydrochloric acid solution equal to 0.3-0.4%, the hydrolysis product has a very small degree of cleavage (up to 5%), which will result in the cultivation of the enzymatic stage of hydrolysis (up to 10 days) and, as a consequence, deterioration of the physical properties of the hydrolyzate — dark brown color,

Example 2. Baseline data: hydrochloric acid concentration 0.3%, hydrolysis duration 20 minutes, temperature 90 C.

300 kg of ground sausage casing wastes are loaded into the reactor, 495 l of water and 3.75 of concentrated hydrochloric acid are added, the hydrolysis of the raw material is carried out for 20 minutes with a 0.3% hydrochloric acid solution. Liquid hydrolysis is dried on a one-way atmospheric dryer. The obtained dry product is subjected to enzymatic hydrolysis using pancreatic minced meat and tap water (at the rate of 1 kg of dry powder, 1 kg of minced pancreatic meat per 10 l of water), alkalinized with 20% 5.MstON to pH 8.0, preserved, added 1.3% chloroform. The hydrolysis is carried out for 8 days under thermostat conditions at 42 s.

Chemical indicators of the obtained base:

Duration of the process, days8 Total nitrogen, mg% 1750 Amine nitrogen, mg% 1138 5 Chlorides,% 0.838 Tryptophan Traces Peptone,% 1.3 Dry residue,% 10.25 This example confirms the base with the required degree of cleavage and chemical indicators , but the time of hydrolysis increased to 8 days, which degrades the physical properties of the base.

5 Due to the fact that the degree of acid hydrolysis of waste, due to temperature and time factors, in this case is less (about 3.8%), the duration of the enzyme. effective hydrolysis, necessary to obtain the base with the desired degree of cleavage, is increased to 8 days.

Example 3. Baseline data. The concentration of Hte is 0.4%, the duration of the hydrolysis is 30 minutes, the temperature of hydrolysis.

300 kg of shredded wastes of tanned sausage casing are loaded into the reactor, 495 liters of water 0 and 5 liters of concentrated saline are added

acids, hydrolysis of RAW MATERIALS is carried out for 30 minutes at 100 ° C in a 0.4% hydrochloric acid solution. The liquid hydrolysis products are dried on a single-roll atmospheric dryer at a pressure of 5 atm and temperature. The resulting dry product is then subjected to enzymatic hydrolysis using minced pancreas and tap water (from

0 calculate 1 kg of dry powder, 1 kg of minced pancreas per 10 l of water), alkalinized once with 20% NaOH to pH 8.0, once preserved by adding 1.3% chloroform. Hydrolysis is carried out for 3 days under thermostat conditions at.

Chemical indicators of the obtained base: 0 Duration of the process, h 72

Total nitrogen, mg% 1500 Amine nitrogen, mg% 1100 Chlorides,% .1.02

TryptophanSledges

5 Peptone,%. 1.08 residual 11 Average final product, l8.5 This example confirms the preparation of a nutritional basis in a short time with the necessary degree of cleavage and physicochemical indicators. With an increase in hydrolysis time up to 50–70 minutes and temperatures up to 120140 s at the same acid concentration (0.3–0.4%), the waste sausage casing is destructed by 15–20%, while the MO hydrolyzate decreases its physical properties, those. it becomes so dark that the nutrient medium prepared on its basis becomes unsuitable for its physical properties. In the case of only acidic hydrolysis using hydrochloric acid to obtain a hydrolyzate with the required degree of cleavage, used as a basis for nutrient media, it is necessary to act on protein waste with strong acid (1.5%), a long time (3.5%). h) at high temperature (130-140 ° C), as a result, the hydrolysis products have poor physical properties (dark brown color), due to the formation of undesirable humic substances under the influence of strong acid, which has a long effect on waste, amino acids are required for the growth of microorganisms (such as tyrosine, arginine, serine, threonine glycyl, alanine, methionine, and others), and hydrolysis products contain significant amounts of acid that must be removed or neutralized. A large amount of mineral salts (ash content), which is unacceptable to be based on nutrient media. The obtained dry hydrolyzate must be further subjected to fiatnal hydrolysis using the io-gastric gland. in connection with the fact that it has a small degree (10-13%) cleavage, inadequate foundations for culture media (557J3%). Moreover, enzymatic hydrolysis is softer and more gentle, does not destroy amino acids. The conditions of the environment in which the hydrolysis occurs are extremely important for the final result. The temperature factor in the hydrolysis is decisive. The authenticity of the enzyme complex of the pancreas drops sharply when it is heated with its aqueous jBbnue solutions. The pancreatic complex of the pancreas, like most enzymes, is extremely sensitive to the effects of temperatures above. The rate of hydrolytic action increases with increasing temperature, according to the general law of chemical kinetics, and this increase ultimately leads to irreversible inactivation of enzymes. The optimum of the final results and hydrolysis are in the temperature range of 40-42 ° C. The duration of hydrolysis for 2-3 days provides the necessary accumulation of amino nitrogen in the hydrolyzate and the desired degree of splitting of the raw material. Similarly, the pH of the medium is set in such a way as to ensure maximum activity of the enzyme used. The optimal response of the medium that ensures the maximum activity of the pancreas at a given temperature of 40-42 ° C is in the range of 7.8-8.0. The pH value can be adjusted to the required limits during the hydrolysis process by adding the appropriate alkaline or acidic substances. With a shorter time of pancreatic action (up to 1 day) on acid hydrolytic, the degree of cleavage of the latter decreases, with a longer (more than 3 day) increase, i.e. in the first case, some high molecular weight peptides and peptones predominate in the hydrolyzate; in the second, low molecular weight peptides and amino acids. Example 4. Initial data HSE concentration 0.4%, hydrolysis duration 30 min, hydrolysis temperature. 300 kg of shredded waste of tanned sausage casing is loaded into the reactor, 495 l of water and 5, l of concentrated hydrochloric acid are added, hydrolysis of the raw material is carried out for 20 minutes at 0.4% hydrochloric acid solution. The liquid hydrolyzate is dried on a single-roll atmospheric drier at a pressure of 5 atm and a temperature of 102 ° C. Semi-dry product is then subjected to fermentative hydrolysis using pancreatic minced meat and tap water (from the calculation of 1 kg of dry powder, 10 liters of odes and 1 kg of minced pancreatic stones, once basified with a 20% NaOH solution to a pH of 7.8, the bottom preserved by adding chloroform. Hydrolysis is allowed to melt for 1 day under thermostat conditions at 40 ° C.

Chemical indicators of the obtained base:

Duration of the process, h.24 Total nitrogen, mg% 1125 Amine nitrogen, mg% 460 Chlorides,% 0.945 Tryptophan Traces Peptone,% 2.5 Dry residue,% 10.04 Average yield, l 8.5 This example confirms receipt product with a small degree of cleavage (up to 40%), which is insufficient for the base.

Example 5. Baseline data: HCB concentration 0.4%, hydrolysis duration 30 min, hydrolysis temperature 100 ° C.

The conditions of acid and fermentative hydrolysis are analogous to Example 4, but the duration of the fermentatious hydrolysis is 6 days.

Chemical indicators of the obtained base:

The duration of the process, Sutb

Total nitrogen, mg% 1600 Amine nitrogen, mg% 1325 Chlorides,% 0.98

TryptophanSledges

Peptone,% 0,92

The dry residue% 13

The average yield of the finished product

This example confirms the production of a product with a high degree of cleavage (more than 80%), which is undesirable for the preparation of the base, since in this case the product contains almost one amino acid and is absent. High-molecular compounds necessary for the growth of bacteria and microorganisms.

For most microbes to grow well, it is necessary that the average amount of amino nitrogen in the nutrient medium, giving an idea of the content of amino acids, is within 25-30% of the total nitrogen content. The hydrolysis time of 2–3 days allows one to obtain such a basis, which can be used to prepare nutrient media with the necessary content of amine and total nitrogen.

These bases are used to prepare nutrient media to control sterility according to the conventional method.

Thus, the advantages of using the proposed base in the production of nutrient media compared with casein are low cost (the base is prepared from waste products, non-food protein, the cost of 1 kg of casein is 2 rubles, the proposed price is 1 rub.) saving chemical reagents (soda, caustic soda, chloroform, etc.) and materials, high yield (85%) of the target product with 1 kg of raw material, compared with casein (65%), which swells, keeps the liquid phase in the sediment, high standard of raw materials amino acid composition, high growth properties of microorganisms and the possibility of centralized production of the base in large volumes.

Save money by using predla-. The average size of the medium base enterprise will be 18 thousand rubles. compared to casein.

Claims (1)

METHOD FOR PRODUCING PROTEIN HYDROLYSIS FROM COLLAGEN-CONTAINING WASTES OF PRODUCTION OF THE PROTEIN Sausage Shell, including hydrolysis of the feedstock, which is based on the fact that, in order to obtain a hydrolyzate suitable for. Preparation of microbiological nutrient media to control sterility, hydrolysis of raw materials is carried out in a 0.4–0.3% hydrochloric acid solution for 30–40 min at. 100-120 ° C, and then subjected to enzymatic hydrolysis, which OSU 'is created using the pancreas, at 40-42 ° C for 2-3 days. _with f