The method of obtaining basics of microbiological culture media

 

(57) Abstract:

The invention relates to biotechnology. Protein-containing raw material is crushed. The extraction is carried out protein from a raw material by mixing it with a solution of Catolica with pH 11,0 - 13,0 with subsequent heating and temperature control. Protein hydrolysis is carried out in the cathode and anode chambers of the electrolyzer at the same time in the countercurrent mode. The neutralization is carried out until pH 5,0 - 8,5 required for optimal growth of microorganisms. The method allows to reduce energy and labor.

The invention relates to biotechnology, and in particular to methods of obtaining drugs and environments for cultivation and detection of microorganisms, and can be used in the microbiological industry, medicine, veterinary science and agriculture.

A method of obtaining nutrition from waste from the processing of krill, by hydrolysis with Pancreatin. /Majidov, M. M., sultans z z the Use of non-food raw materials in the production of microbiological culture media/ Institute for production pitt. environments. - Makhachkala: Doug.kN.publishing house. 1986, S. 45-47/

The drawbacks are the use of scarce and expensive pancreatic enzymes, niscom decision to declare adopted for the prototype of the invention is a method of obtaining a protein hydrolysate from hydrobionts (A. C. USSR N 1687213, CL A 23 J 1/4, 1991), according to which the hydrolytic extraction of the protein from the crushed material and neutralizing the solution is carried out in a diaphragm electrolyzer. The conditions of alkaline hydrolysis is provided by passing a constant current density of 300-600 a/m20.5 - 2.0% NaCl solution in the cathode chamber of the electrolytic cell with subsequent heating to 80-90oC, and the neutralization is carried out in the anode chamber of the electrolyzer at the same current density until reaching neutral pH values.

The disadvantages of the prototype are the low degree of protein hydrolysis, which is characterized by a content of amino nitrogen is 1.2%, the low yield of the target product is 40-80%, greater power consumption and correspondingly high cost of the product.

The invention is directed to a technology for obtaining basics of microbiological culture media for various purposes from protein-containing raw materials, including waste from cutting marine and freshwater crustaceans.

This solved the problem of creating an efficient method for the extraction and hydrolysis of protein with a high in the proposed method, including the grinding of raw materials, extraction and hydrolysis of the protein, in contrast to the prototype extraction of protein from shredded protein-containing raw material carried by mixing with a solution of Catolica with a pH value of 11.0-13.0, followed by heating and temperature control.

The catholyte is a solution produced in the cathode chamber of a diaphragm electrolytic cell in the electrolysis conductive media DC electric field. In this case, as Catolica used solution is waste in the process of obtaining chitin from crab by-products raw materials by electrochemical method (A. S. USSR N 1751888, CL A 23 L 1/33, 1990), implemented on the same equipment and in the same process cycle, and that the inventive method. It is formed when deproteinizovana shell in the presence of an electrolyte (0.5-5.0% solution of NaCl or Na2SO4in the electrolysis process of this suspension in a cathode chamber of the electrolytic cell.

Hydrolysis of extracted protein is carried out in the cathode and anode chambers of diaphragm electrolyzer simultaneously in the mode of counter-current at a current density of 200-650 a/m2. The process of electrobraid protein solution is performed in the cathode, and then in the anode Kam is achieving in the anode chamber pH 5.0-8.5, which is the optimum for the growth of microorganisms, which should coincide with the formation in a mixture of easily separated liquid, translucent fraction and a solid residue.

The proposed method was implemented in the laboratory in the double-cell, separated by a diaphragm to the anode and the cathode space, with the electrodes connected respectively with the positive and negative poles of the DC source.

The essence of the proposed method is illustrated by the following examples.

Example 1

Raw - dried gammarus, mixed with water in the ratio 1:1 and grinded on any dispersing equipment to the size of the particles < 510-3m, then was soaked in catholyte in the ratio 1:2 and was maintained for at least 3 hours at room temperature. (Catholyte was a waste of technology for chitin and was formed during the filtration of a suspension of the shell with 1% NaCl (in the ratio 1:20), held electrochemical processing in a cathode chamber of a diaphragm electrolytic cell at a current density of 220 a/m2within 30 minutes, and temperature-controlled at 70+5oC for 30 minutes under stirring.) Suspension catholyte : Serra by filtration. The protein solution for the secondary hydrolysis was applied to electrobraid to the cathode diaphragm electrolyzer. Treatment of the protein solution was carried out at current density of 220 a/m2to achieve pH 11.0. The protein solution passed cathodic treatment, was fed to the anode chamber of the electrolyzer for additional hydrolysis and have kind of balanced out at current density of 220 a/m2to pH 7.4, which is optimal for bacteria, for example: Bacillus mucilaginosus, Escherichia coli. Formed in the hydrolysis of the precipitate was separated by centrifugation. The obtained protein hydrolyzate was a transparent brown solution with a content of amino nitrogen - 2.2%, protein - 73%, NaCl - 17%.

Example 2

The experiment was carried out analogously to example 1, the raw materials were mixed with Catolica in the ratio of 1:5, electrobraid protein solution in the cathode chamber was carried out to the value pH12.0 current density 430 a/m2. The obtained protein hydrolyzate was a transparent brown solution with a content of amino nitrogen - 2.8%, protein - 59%, NaCl - 20%.

Example 3

The experiment was carried out analogously to example 1, the raw materials were mixed with Catolica in the ratio of 1: 10, electrobraid belovedtales a clear solution light brown colour with the content of amino nitrogen - 2.5% protein and 44%, NaCl - 32%.

Example 4

Raw krill, frozen. Analogously to example 1, but there is no stage of mixing the raw material with water before dispersing. Processing of protein solution in the cathode chamber was carried out up to pH 13.0 with current density 540 a/m2. The obtained protein hydrolyzate was a clear solution light brown color with a content of amino nitrogen 2.8% protein, 57%, NaCl - 25%.

Analysis of the obtained results allows to conclude that when using a current of less than 200 a/m2not achieved the maximum possible degree of protein hydrolysis, and at current density of more than 600 a/m2there is a strong heating of the camera, which may lead to deformation of the membrane and lower quality protein solution. When the water ratio of raw materials: the catholyte less than 1:2 protein solution contains a high concentration of NaCl, and when the water ratio more than 1:10 is not possible for full extraction of protein from raw materials.

Obtained as described in examples 2,4 protein hydrolysates from gammarus and krill used as bases microbiological nutrient media for growing bacteria Bacillus mucilaginosus, producer of biologically active substances.

Cultivatin. The volume of the medium 50 cm3the hydrolysate gammarus is 96% molasses 4%. Exit completely dry biomass amounted to 8.4 - 9.6 g/DM3. When using krill hydrolysate exit completely dry biomass amounted to 8.8 - 10.8 g/DM3.

The method of obtaining basics of microbiological culture media, including the grinding of raw materials, extraction and protein hydrolysis, neutralization and separation of nerastvorimogo sludge, characterized in that the extraction of protein from raw materials carried out by mixing the crushed material with a solution of Catolica with pH 11,0 - 13,0, taken in an amount of 2 to 10 wt.h. 1 wt.h. raw materials, and temperature control, hydrolysis are processed in the cathode and anode chambers of diaphragm electrolyzer at the same time in the countercurrent mode, and the neutralization is carried out in the anode chamber simultaneously with the hydrolysis until a pH of 5.0 to 8.5.

 

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