RU2416633C2 - Method for production of hydrolysate of californian worms - Google Patents

Abstract

FIELD: food industry.

SUBSTANCE: method envisages blending worms biomass with tap water at a weight ratio of 1:2 respectively with subsequent boiling during 8-10 minutes. Worms biomass is separated from broth and milled in a homogeniser to produce mince. The broth is cooled to a temperature of 45-50°C and alkalised with Na₂CO₃ to pH 8.2 in terms of phenolphthalein. The Californian worms mince is blended with the broth and added cattle pancreas to in an amount of 40-150 g/l. 2% of chloroform is added to the total volume which is then maintained at a temperature of 37°C during 14 days. One shakes the mass every 15 minutes for 5 minutes during the first day and every 2 hours for 5 minutes on subsequent days. One filters the mass and adds 2% of chloroform to the filtrate.

EFFECT: invention allows to improve the hydrolysate quality and simplify its production process.

2 tbl, 6 ex

Description

FIELD OF THE INVENTION

The invention relates to biotechnology, in particular to methods for producing a hydrolyzate from California worms, and can be used as a biologically active component of food additives and nutrient media for the cultivation of microorganisms.

State of the art

A known method of producing an enzymatic hydrolyzate, including the processing of protein-containing raw materials diluted with water with medical pancreatin or native pig pancreas, in an alkaline zone of pH 7.6-7.8 in the presence of chloroform at 37-38 ° C for 6-7 days, heating at 90 ° C for 5-10 min, subsequent precipitation of high molecular weight compounds at the isoelectric point of the protein, filtering the target product and drying it, while fat-free, dehydrated brain tissue is used as protein-containing raw materials cattle, the enzyme is dissolved in water in a ratio of 1:25 and before precipitation of high molecular weight compounds, the hydrolyzate is treated with a 20-22% solution of calcium chloride for 5-10 minutes (see US Pat. RU No. 2020153, IPC C12N 5/00 , С12Р 21/06, published on September 30, 1994).

The disadvantage of this method is the complexity of the process, the high cost of the final product.

A known method of producing protein hydrolyzate from meat and meat and bone raw materials of slaughter animals, which consists in the fact that the crushed meat and meat and bone raw materials are homogenized with water and degreased. Enzymatic hydrolysis is carried out by a complex of proteolytic enzymes obtained from the pancreas of slaughter animals and protosubtilin G20x. Horned hoof flour is additionally used as raw material, and the fat fraction obtained after degreasing is subjected to fermentation. Biomass is mixed with meat and bone broth and blood of slaughtered animals in equal proportions, incubated for 20-24 hours at a temperature of 45-50 ° C. The horned flour is subjected to acid hydrolysis with a solid fraction for 5-6 hours at a temperature of 110-120 ° C, followed by cooling and drying. Fermentation of the fat fraction is carried out by bacterial culture of the yeast Candida tropicalis strain VSB-28 in the presence of mineral salts and at a temperature of 37-40 ° C for 6-8 hours with continuous aeration. The method allows to increase the degree of protein conversion and use in the process all the waste from the meat industry (see US Pat. RU No. 2132142, IPC A23J 1/10, A23J 3/00, publ. 06/27/1999).

The disadvantage of this method of obtaining protein hydrolyzate is the high cost of the final product.

The closest in technical essence and the achieved positive effect and adopted by the authors for the prototype is a method of producing a hydrolyzate of the following composition g / l: beef or horse meat is cleaned of fat and tendons, cut into slices or stripes in the thickness of a finger. Put in boiling water at the rate of 2 liters of water per 1 kg of meat; boil for 5 minutes. The meat is caught and passed through a meat grinder. The minced meat is mixed with a liquid and cooled to 45 ° C, after which the mixture is placed in a bottle with a tight rubber stopper, to which 5-10 g of pancreatin or 40-80 g of the pancreas are added (depending on what degree of splitting they want to get) (Kozlov Yu . A. Nutrient media in medical microbiology / Yu.A. Kozlov, 1950. - P. 51).

The disadvantage of this hydrolyzate is the high cost.

Disclosure of invention

The objective of the invention is to develop a method for producing a hydrolyzate from California worms with high quality and low cost.

The technical result that can be obtained using the proposed invention is to improve the quality and reduce the cost.

The technical result is achieved using a method of producing a hydrolyzate from California worms, which involves mixing the biomass of the worms with tap water in a ratio of 1: 2, respectively, boiling for 8-10 minutes, separating the biomass of the worms from the broth, grinding it on a homogenizer to produce minced meat, and cooling the broth to 45-50 ° C, alkalinization broth Na ₂ CO ₃ to pH 8.2 to phenolphthalein, mixing minced Californian worms broth, adding the pancreas of cattle in an amount of 40-150 g / l, was added the total volume of 2% chloroform, keeping for 14 days at a temperature of 37 ° C, shaking during the first day every 15 minutes for 5 minutes, and on the following days - every 2 hours for 5 minutes, filtering, adding 2 % chloroform in the filtrate.

The essence of the preparation of enzymatic hydrolyzate from California worms is as follows. The biomass of California worms in an amount of 0.5 kg is combined with 1 liter of tap water and boiled for 8-10 minutes. Then the worms are separated from the broth and ground in a homogenizer to a homogeneous mass. The broth is cooled to a temperature of 45-50 ° C, alkalized with Na ₂ CO ₃ to a pH of 8.2 by phenolphthalein. Stuffed California worms are placed in a three-liter glass container, poured with prepared cooled broth, 150 g of cattle pancreas are added. 2% chloroform is poured to the total volume of the contents of the balloon, covered with a tight cotton-gauze swab with parchment. The cylinder is placed in a heat chamber at a temperature of 35-37 ° C.

It is aged for 14 days, shaken during the first day every 15 minutes for 5 minutes, and on the following days every two hours for 5 minutes. The level of amino nitrogen is measured daily. At the end of the digestion, the liquid is filtered through a Belting cloth and 4 layers of filter paper. Amine nitrogen is measured. 2% chloroform is added to the filtrate, poured and stored at a temperature of 2 to 8 ° C. The hydrolyzate is a cognac-colored liquid.

The implementation of the invention

Examples of specific performance of the method of obtaining a hydrolyzate from California worms.

Example 1. The biomass of California worms contains up to 82% protein, therefore, is a protein-containing raw material. Previously, the biomass of California worms is washed - this is the preparation of protein-containing raw materials. Then, 0.5 kg biomass is combined with 0.5 l of tap water and boiled for 8 minutes. Then the worms are separated from the broth and ground in a homogenizer to a homogeneous mass to obtain minced meat. The broth is cooled to a temperature of 45-50 ° C, alkalized with Na ₂ CO ₃ to a pH of 8.2 by phenolphthalein. Stuffed California worms are placed in a three-liter glass container, poured with prepared cooled broth, 40 g of cattle pancreas are added. 2% chloroform is poured to the total volume of the contents of the balloon, covered with a tight cotton-gauze swab with parchment. The cylinder is placed in a heat chamber at a temperature of 35 ° C. It is aged for 14 days, shaken during the first day every 15 minutes for 5 minutes, and on the following days every two hours for 5 minutes. The level of amino nitrogen is measured daily. At the end of the digestion, the liquid is filtered through a Belting cloth and 4 layers of filter paper. Amine nitrogen is measured. 2% chloroform is added to the filtrate, poured and stored at a temperature of 2 to 8 ° C. The hydrolyzate is a cognac-colored liquid.

The resulting base has a precipitate, poorly filtered.

Example 2. The biomass of California worms in the amount of 0.5 kg is combined with 1 l of tap water and boiled for 10 minutes. Then the worms are separated from the broth and ground in a homogenizer to a homogeneous mass to obtain minced meat. The broth is cooled to a temperature of 45-50 ° C, alkalized with Na ₂ CO ₃ to a pH of 8.2 by phenolphthalein. Stuffing of California worms is placed in a three-liter glass container, filled with ready-made cooled broth, 80 g of cattle pancreas are added. 2% chloroform is poured to the total volume of the contents of the balloon, covered with a tight cotton-gauze swab with parchment. The cylinder is placed in a heat chamber at a temperature of 37 ° C. It is aged for 14 days, shaken during the first day every 15 minutes for 5 minutes, and on the following days every two hours for 5 minutes. The level of amino nitrogen is measured daily. At the end of the digestion, the liquid is filtered through a Belting cloth and 4 layers of filter paper. Amine nitrogen is measured. 2% chloroform is added to the filtrate, poured and stored at a temperature of 2 to 8 ° C. The hydrolyzate is a cognac-colored liquid.

With this hydrolysis, incomplete breakdown of proteins to amino acids occurs, which reduces the biological value.

Example 3. The biomass of California worms in the amount of 0.5 kg is combined with 1 l of tap water and boiled for 10 minutes. Then the worms are separated from the broth and ground in a homogenizer to a homogeneous mass to obtain minced meat. The broth is cooled to a temperature of 45-50 ° C, alkalized with Na ₂ CO ₃ to a pH of 8.2 by phenolphthalein. Stuffed California worms are placed in a three-liter glass container, poured with prepared cooled broth, 100 g of cattle pancreas are added. 2% chloroform is poured to the total volume of the contents of the balloon, covered with a tight cotton-gauze swab with parchment. The cylinder is placed in a heat chamber at a temperature of 37 ° C. It is aged for 14 days, shaken during the first day every 15 minutes for 5 minutes, and on the following days every two hours for 5 minutes. The level of amino nitrogen is measured daily. At the end of the digestion, the liquid is filtered through a Belting cloth and 4 layers of filter paper. Amine nitrogen is measured. 2% chloroform is added to the filtrate, poured and stored at a temperature of 2 to 8 ° C. The hydrolyzate is a cognac-colored liquid.

With this hydrolysis, incomplete breakdown of proteins to amino acids occurs.

Example 4. The biomass of California worms in the amount of 0.5 kg is combined with 1 l of tap water and boiled for 10 minutes. Then the worms are separated from the broth and ground in a homogenizer to a homogeneous mass to obtain minced meat. The broth is cooled to a temperature of 45-50 ° C, alkalized with Na ₂ CO ₃ to a pH of 8.2 by phenolphthalein. Stuffed California worms are placed in a three-liter glass container, poured with prepared cooled broth, 150 g of cattle pancreas are added. 2% chloroform is poured to the total volume of the contents of the balloon, covered with a tight cotton-gauze swab with parchment. The cylinder is placed in a heat chamber at a temperature of 37 ° C. It is aged for 14 days, shaken during the first day every 15 minutes for 5 minutes, and on the following days every two hours for 5 minutes. The level of amino nitrogen is measured daily. At the end of the digestion, the liquid is filtered through a Belting cloth and 4 layers of filter paper. Amine nitrogen is measured. 2% chloroform is added to the filtrate, poured and stored at a temperature of 2 to 8 ° C. The hydrolyzate is a cognac-colored liquid.

The resulting hydrolyzate is well filtered, has a high biological activity, proteins are cleaved to amino acids as much as possible.

Example 5. The biomass of California worms in the amount of 0.5 kg is combined with 1 l of tap water and boiled for 10 minutes. Then the worms are separated from the broth and ground in a homogenizer to a homogeneous mass to obtain minced meat. The broth is cooled to a temperature of 45-50 ° C, alkalized with Na ₂ CO ₃ to a pH of 8.2 by phenolphthalein. Stuffed California worms are placed in a three-liter glass container, poured with prepared cooled broth, 180 g of cattle pancreas are added. 2% chloroform is poured to the total volume of the contents of the balloon, covered with a tight cotton-gauze swab with parchment. The cylinder is placed in a heat chamber at a temperature of 37 ° C. It is aged for 14 days, shaken during the first day every 15 minutes for 5 minutes, and on the following days every two hours for 5 minutes. The level of amino nitrogen is measured daily. At the end of the digestion, the liquid is filtered through a Belting cloth and 4 layers of filter paper. Amine nitrogen is measured. 2% chloroform is added to the filtrate, poured and stored at a temperature of 2 to 8 ° C. The hydrolyzate is a cognac-colored liquid.

With this formulation, the hydrolyzate of example No. 5 is not inferior in biological value to the hydrolyzate prepared according to example No. 4. The disadvantage of the latter hydrolyzate is the use of a larger amount of cattle pancreas, which, when converted to economic efficiency, increases money costs.

Thus, the most optimal and relevant requirements for hydrolysates is the method of producing hydrolyzate according to example No. 4. The hydrolyzate has a cognac color, is well filtered and has a high biological activity.

Example No. 6. To confirm the biological activity of a hydrolyzate from California worms prepared according to example No. 4, we conducted a study of its chemical composition, tested for activity by the method of K. Bakirdzhiev (I. Kalashnik Stimulating therapy in veterinary medicine / I. A. Kalashnik. - 2nd ed., Revised and supplemented - K .: Harvest, 1990. - 160 p.), And also conducted a study of the hydrolyzate as the basis of microbiological culture media for increasing the bacterial mass. For this, the meat hydrolyzate in the Hottinger agar was replaced with a hydrolyzate from California worms according to example No. 4.

A chemical study of the obtained hydrolyzate was carried out for the presence of amino acids, carbohydrates, as well as micro and macro elements.

Based on the studies obtained the following results are table 1.

Table 1 Some chemical components of the hydrolyzate from California worms, prepared according to example No. 4 Chemical components / quality indicators Quantitative indicators 1. Amino acids: g / l: threonine 51.1 ± 0.2 proline 41.2 ± 0.1 methionine 23.3 ± 0.8 leucine 38.0 ± 1.0 hydrochloric arginine 43.0 ± 0.5 2. Carbohydrates: - glucose the quantity was not calculated traces of lactose the quantity was not calculated glycogen the quantity was not calculated 3. Micro and macro elements mg / l: zinc 4.9 iron 38,4 sodium 29.3 potassium 245 magnesium 28.7 calcium 19.7

Determination of hydrolyzate activity by the method of K. Bakirdzhiev.

Take 2 bottles of hydrolyzate. 500 mg of yeast, 19.5 ml of glucose and 0.5 ml of 0.9% sodium chloride solution are placed in the first tube (control).

In the second and third test tubes - 500 mg of yeast, 19.5 ml of glucose, 0.5 ml of hydrolyzate. The tubes are tightly corked with rubber stoppers with curved glass tubes inserted into them, through which liquid is displaced, displaced by carbon dioxide produced by the yeast during their life. The volume of displaced fluid in the tubes is determined every 24 hours.

The difference in the volume of displaced fluid between the control tube and the test was 30%. This indicates a high biological activity of the hydrolyzate.

In the study of the hydrolyzate as the basis of microbiological culture media for increasing the bacterial mass, cultures of test strains are tested: Y. pestis Ev; P. aeruginosa 27/99; S.marcescens-1; lactobacterii-cn; E. coli CA-18. The growth of microorganisms on a nutrient medium containing a hydrolyzate from California worms as a nutrient base exceeds the growth of microorganisms on the Hottinger agar. In the new nutrient medium, continuous growth with the formation of intense red pigment for S.marcescens-1 and continuous growth with the formation of intense blue-green pigment for P. aeruginosa 27/99, as well as an increase in the number of grown colonies for E. coli CA-18, are observed. Y. pestis EV, Lactobacterii-cn. The data are presented in table 2.

Thus, the developed method for producing a hydrolyzate from California worms with minimal costs allows you to save a wide range of biological components, which gives the right to use it in various industries.

The invention in comparison with the prototype and other known technical solutions has the following advantages.

The hydrolyzate from California worms surpasses traditional types of hydrolysates (in particular, meat and sprats) in the following parameters:

- includes a large number of highly active low molecular weight nitrogen compounds;

- contains a variety of macro- and microelements: Zn, Fe, Na, K, Mg, Ca;

- environmental cleanliness, lack of antibiotics and other harmful impurities;

- has a lower cost.

Claims (1)

A method of producing a hydrolyzate from California worms, comprising mixing the biomass of the worms with tap water in a ratio of 1: 2, respectively, boiling for 8-10 minutes, separating the biomass of the worms from the broth, grinding it on a homogenizer to produce minced meat, cooling the broth to 45-50 ° C, alkalization of Na ₂ CO ₃ broth to pH 8.2 in phenolphthalein, mixing mince of California worms with broth, addition of cattle pancreas in an amount of 40-150 g / l, addition of 2% chloroform to the total volume, keeping for 14 days at a temperature of 37 ° C, shaking during the first day every 15 minutes for 5 minutes, and on the following days every 5 hours for 5 minutes, filtering, adding 2% chloroform to the filtrate.