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Method for releasing intracellular enzyme in bacterial culture

A technology of bacterial culture and intracellular enzymes, applied in the direction of microorganism-based methods, bacteria, biochemical equipment and methods, etc., can solve the problems of destroying enzyme activity, solvent residue, high cost, etc., to achieve improved enzyme activity, no solvent residue, The effect of increasing content

Pending Publication Date: 2022-04-29
北京四良科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Solve technical problems such as destruction of enzyme activity, high cost, and solvent residue during the release of intracellular enzymes

Method used

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  • Method for releasing intracellular enzyme in bacterial culture
  • Method for releasing intracellular enzyme in bacterial culture

Examples

Experimental program
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Effect test

Embodiment 1

[0025] A method for bacterial culture to release intracellular enzymes, characterized in that: the method comprises the following steps:

[0026] (1) Under aseptic conditions, pick the strains from the bacillus strain preservation slope, insert them into the liquid medium, and shake at 30°C for 24 hours at a rotational speed of 180r / min;

[0027] (2) When the bacillus is cultivated to a certain OD value, it is subjected to stress culture to rupture the bacterium and release the intracellular enzyme;

[0028] Further, the bacillus is bacillus subtilis;

[0029] Further, the OD value in the step (2) is 6;

[0030] Further, the stress cultivation method is as follows: stop oxygen supply, cut off oxygen, cultivate at 50° C. for 20 minutes, add 5% ammonium chloride, and continue to cultivate for 5 hours.

[0031] Further, the liquid medium in the step (1) is: 10 g of peptone, 5 g of yeast extract, 10 g of sodium chloride, 1000 mL of distilled water, pH 7.0-7.2, and sterilized at ...

Embodiment 2

[0033] A method for bacterial culture to release intracellular enzymes, characterized in that: the method comprises the following steps:

[0034] (1) Under aseptic conditions, pick the strains from the bacillus strain preservation slope, insert them into the liquid medium, and shake at 30°C for 24 hours at a rotational speed of 180r / min;

[0035] (2) When the bacillus is cultivated to a certain OD value, it is subjected to stress culture to rupture the bacterium and release the intracellular enzyme;

[0036] Further, the bacillus is bacillus licheniformis;

[0037] Further, the OD value in the step (2) is 6;

[0038] Further, the stress cultivation method is as follows: stop oxygen supply, cut off oxygen, cultivate at 50° C. for 20 minutes, add 5% ammonium sulfate, and continue to cultivate for 5 hours.

[0039] Further, the liquid medium in the step (1) is: 10 g of peptone, 5 g of yeast extract, 10 g of sodium chloride, 1000 mL of distilled water, pH 7.0-7.2, and sterilized...

Embodiment 3

[0041] A method for bacterial culture to release intracellular enzymes, characterized in that: the method comprises the following steps:

[0042] (1) Under aseptic conditions, pick the strains from the bacillus strain preservation slope, insert them into the liquid medium, and shake at 30°C for 24 hours at a rotational speed of 180r / min;

[0043] (2) When the bacillus is cultivated to a certain OD value, it is subjected to stress culture to rupture the bacterium and release the intracellular enzyme;

[0044] Further, the bacillus is bacillus subtilis;

[0045] Further, the OD value in the step (2) is 8;

[0046] Further, the stress cultivation method is as follows: stop oxygen supply, cut off oxygen, cultivate at 50° C. for 20 minutes, add 5% ammonium chloride, and continue to cultivate for 5 hours.

[0047] Further, the liquid medium in the step (1) is: 10 g of peptone, 5 g of yeast extract, 10 g of sodium chloride, 1000 mL of distilled water, pH 7.0-7.2, and sterilized at ...

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Abstract

The invention relates to a method for releasing intracellular enzyme during bacterial culture, which comprises the following steps: under a sterile condition, selecting a ring from a bacillus strain preservation slope, inoculating the ring into a liquid culture medium, carrying out shaking culture at 30-37 DEG C at the rotating speed of 180r / min for 24-48 hours, and when bacillus is cultured to a certain OD value, carrying out stress treatment on the bacillus to break thalli, so as to obtain a bacterial strain; therefore, the intracellular enzyme is released. According to the method, various technical problems of enzyme activity damage, high cost, solvent residue and the like in the intracellular enzyme release process are effectively solved, and rapid and efficient release of the intracellular enzyme is realized. The bacterial culture solution rich in intracellular enzyme can efficiently degrade lignin, promote conversion of nitrogen and carbon, quickly degrade organic matters and refractory substances such as lignocellulose raw materials, and can be used for fermenting and degrading agricultural wastes; and the strain can also be synergistically acted with zymophyte and extracellular enzyme to ferment and compost straws, rotten vegetables, garden branches of fruits and vegetables, kitchen garbage and the like and feed the straws.

Description

technical field [0001] The application relates to the field of preparation of agricultural waste starter, in particular to a method for releasing intracellular enzymes through bacterial culture. Background technique [0002] Agricultural wastes are often fermented and reused as resources. However, the common agricultural waste fermentation technology cannot effectively decompose high lignin and high nitrogen raw materials. The fermentation time of raw materials is long, and the synthesis of humic acid macromolecules cannot be precisely controlled, so it cannot be effectively solved. The problems of high carbon and nitrogen gas emissions, low compost yield and low humic acid content in compost occurred during the microbial decomposition process. [0003] At present, the use of enzyme-bacteria compound preparations to ferment agricultural waste to obtain renewable resources is a common technical means in this field, but the preparation methods of various enzyme preparations ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/56C12N9/08C05F17/20C05F5/00C05F9/00C05F11/00C05F15/00A23K10/12A23K10/14A23K10/37C12R1/125C12R1/10
CPCC12N1/20C12N9/54C12N9/0065C05F17/20C05F11/00C05F5/002C05F9/00A23K10/12A23K10/14A23K10/37C12Y304/24082C12Y304/24812C12Y111/01Y02A40/20Y02P60/87Y02W30/40
Inventor 于家伊张文杨明哲任忠秀刘墨
Owner 北京四良科技有限公司
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