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Protecting agent for stability of marine protease-producing bacillus

A technology for producing protease and bacillus, applied in the direction of preservation of microorganisms, microorganisms, methods based on microorganisms, etc., can solve the problems of low survival rate and decreased activity

Active Publication Date: 2019-10-01
威海长青海洋科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Since this kind of protease-producing bacillus comes from the ocean, its growth and survival conditions should be similar to the marine ecological environment, but usually the strains cultured under the environmental conditions simulated in the laboratory will have low survival rate and decreased activity when stored in cold storage. question

Method used

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  • Protecting agent for stability of marine protease-producing bacillus
  • Protecting agent for stability of marine protease-producing bacillus
  • Protecting agent for stability of marine protease-producing bacillus

Examples

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Effect test

Embodiment 1

[0013] A marine protease-producing bacillus stability protection agent, which is obtained by adding glycerin, mannitol, EDTA, potassium chloride, magnesium sulfate, disodium hydrogen phosphate, and diphenyl antimony 2-ethylhexanoate to 10 mM Tris- In a solution of HCl buffer (pH 7.5), stir evenly at room temperature to completely dissolve the solute. After standing still for 10 min, use a solution of 10 mM Tris-HCl buffer (pH 7.5) to obtain the final protective agent. The concentrations of the ingredients are: 32% Glycerin (v / v), 5.50% Mannitol (w / v), 1.50% EDTA (w / v), 0.55% Potassium Chloride (w / v), 0.35% Magnesium Sulfate ( w / v), 0.32% disodium hydrogen phosphate (w / v), 0.05% diphenylantimony 2-ethylhexanoate (v / v). The mM unit means millimoles per liter.

[0014] Take the live bacterial liquid of marine protease-producing Bacillus with a viable bacterial content of 5 billion cfu / mL, add 5% stability protection agent to the bacterial liquid, store it at 4°C for 30 days, tak...

Embodiment 2

[0017] A marine protease-producing bacillus stability protection agent, which is obtained by adding glycerin, mannitol, EDTA, potassium chloride, magnesium sulfate, disodium hydrogen phosphate, and diphenyl antimony 2-ethylhexanoate to 10 mM Tris- In a solution of HCl buffer (pH 7.5), stir evenly at room temperature to completely dissolve the solute. After standing still for 10 min, use a solution of 10 mM Tris-HCl buffer (pH 7.5) to obtain the final protective agent. The concentrations of the ingredients are: 35% glycerin (v / v), 6.2% mannitol (w / v), 3.20% EDTA (w / v), 0.50% potassium chloride (w / v), 0.47% magnesium sulfate ( w / v), 0.30% disodium hydrogen phosphate (w / v), 0.06% diphenylantimony 2-ethylhexanoate (v / v), fixed with a solution of 10 mM Tris-HCl buffer (pH 7.5) Allow. The mM unit means millimoles per liter.

[0018] Take the live bacterial liquid of marine protease-producing Bacillus with a viable bacterial content of 5 billion cfu / mL, add 5% stability protection ...

Embodiment 3

[0021] A marine protease-producing bacillus stability protection agent, which is obtained by adding glycerin, mannitol, EDTA, potassium chloride, magnesium sulfate, disodium hydrogen phosphate, and diphenyl antimony 2-ethylhexanoate to 10 mM Tris- In a solution of HCl buffer (pH 7.5), stir evenly at room temperature to completely dissolve the solute. After standing still for 10 min, use a solution of 10 mM Tris-HCl buffer (pH 7.5) to obtain the final protective agent. The concentrations of the ingredients are: 27% Glycerin (v / v), 5.8% Mannitol (w / v), 1.8% EDTA (w / v), 0.60% Potassium Chloride (w / v), 0.55% Magnesium Sulfate ( w / v), 0.45% disodium hydrogen phosphate (w / v), 0.07% diphenylantimony 2-ethylhexanoate (v / v), fixed with a solution of 10 mM Tris-HCl buffer (pH 7.5) Allow. The mM unit means millimoles per liter.

[0022] Take the live bacterial liquid of marine protease-producing Bacillus with a viable bacterial content of 5 billion cfu / mL, add 5% stability protection a...

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Abstract

The invention relates to a protecting agent for the stability of a marine protease-producing bacillus, and belongs to the technical field of microorganisms. The protecting agent comprises a Tris-HCl buffer solution, glycerol, mannitol, EDTA, potassium chloride, magnesium sulfate, disodium hydrogen phosphate and 2-ethylhexanoic acid diphenyl antimony, wherein the Tris-HCl buffer solution, glycerol,mannitol, EDTA, potassium chloride, magnesium sulfate, disodium hydrogen phosphate and 2-ethylhexanoic acid diphenyl antimony are prepared in a certain ratio into the liquid protecting agent. The protecting agent for the stability of the marine protease-producing bacillus solves the problems that a marine protease-producing bacillus strain is denatured under the low-temperature refrigeration condition, and the survival rate is low, and the survival rate of the strain can be stabilized at 95% or above.

Description

technical field [0001] The invention relates to the field of microorganism preservation, in particular to a protective agent for the stability of marine protease-producing bacillus. Background technique [0002] With the rapid development of large-scale and intensive aquaculture, aquaculture diseases occur frequently. In recent years, people have begun to try to use beneficial microorganisms in breeding production, especially the enzyme-producing probiotics in the ocean, which can improve the ecological environment of farming, improve the immunity of farmed animals, inhibit pathogenic microorganisms, and reduce the occurrence of diseases. Among them, Bacillus, which is derived from marine protease, has been developed in a variety of ways due to its unique protease-producing properties. It can be directly added to aquaculture water or added to feed, and can also be used in mariculture through bait organisms. Animals can also be used by injection or dipping. [0003] Since t...

Claims

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Application Information

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IPC IPC(8): C12N1/04C12R1/07
CPCC12N1/04
Inventor 颜琳姚艳艳常丽荣闫欣丁建姿王振华
Owner 威海长青海洋科技股份有限公司
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