Bacillus subtilis culture medium for producing anti-microbial peptide and application thereof

A Bacillus subtilis culture medium technology, which is applied in the field of Bacillus subtilis culture medium, can solve the problems of energy waste and environmental pollution, and achieve the effects of reducing production costs, high titer and high quantity

Pending Publication Date: 2015-09-02
HUNAN AGRICULTURAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Yellow pulp water contains a large amount of soybean oligosaccharides, protein and other growth factors. The random discharge of

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] (1) Activation culture: Inoculate the bacillus preserved in the slant into the BHI medium for activation, and cultivate it at 30-42° C. and at a rotation speed of 100-250 rpm for 20-24 hours to obtain the activated Bacillus subtilis fermentation broth.

[0020] (2) Seed tank cultivation: 50L seed medium is loaded into a 100L fermenter, and the activated shake flask fermentation liquid is inoculated according to 0.5% to 3% of the total volume of the seed medium, and the fermentation conditions of the control seed tank are: The fermentation temperature is 35-42°C, the rotation speed is 100-400rpm, and the cultivation is carried out for 16-24 hours. The initial pH value before inoculation is 7.0-7.5, and the ventilation rate is 1:0.5.

[0021] (3) The composition of the seed medium is as follows (percentage by weight): 1.5% soluble starch, 1.0% polyvalent peptone, 0.5% sodium chloride, 0.3% potassium dihydrogen phosphate, 0.3% dipotassium hydrogen phosphate, yellow pulp wat...

Embodiment 2

[0025] (1) Activation culture: Inoculate the bacillus preserved in the slant into the BHI medium for activation, and cultivate it at 30-42° C. and at a rotation speed of 100-250 rpm for 20-24 hours to obtain the activated Bacillus subtilis fermentation broth.

[0026] (2) Seed tank cultivation: 50L seed medium is loaded into a 100L fermenter, and the activated shake flask fermentation liquid is inoculated according to 0.5% to 3% of the total volume of the seed medium, and the fermentation conditions of the control seed tank are: The fermentation temperature is 35-42°C, the rotation speed is 100-400rpm, and the cultivation is carried out for 16-24 hours. The initial pH value before inoculation is 7.0-7.5, and the ventilation rate is 1:0.5.

[0027] (3) The composition of the seed culture medium is as follows (percentage by weight): soluble starch 0.5%, polyvalent peptone 0.5%, sodium chloride 0.1%, potassium dihydrogen phosphate 0.1%, dipotassium hydrogen phosphate 0.1%, yellow ...

Embodiment 3

[0031] The difference between this embodiment and embodiment 1 is:

[0032] The composition of the seed culture medium is as follows (percentage by weight): 0.8% of soluble starch, yellow pulp water as the balance, stirred evenly, and heated. Preparation of the yellow pulp water: centrifuge the fresh yellow pulp water, and take the supernatant as the raw material of the liquid medium.

[0033] The fermentation medium is composed of the following raw materials (percentage by weight): 1.5% soluble starch, 0.5% sodium chloride, 0.3% dipotassium hydrogen phosphate, 0.15% magnesium sulfate, 0.015% calcium carbonate, 0.05% manganese sulfate, yellow pulp water As the balance, the pH was adjusted to 7.0 to 7.5.

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Abstract

The invention provides a bacillus subtilis culture medium for producing anti-microbial peptide and application thereof in producing the anti-microbial peptide. The fermentation medium is composed of, by weight, 0.5%-1.5% of soluble starch or corn starch, 0%-1.0% of polyvalent peptone or beef extract, 0%-0.5% of sodium chloride, 0%-0.3% of monopotassium phosphate, 0%-0.3% of dipotassium phosphate, 0.05%-0.15% of magnesium sulfate, 0.005-0.015% of calcium carbonate, 0.05%-0.15% of manganese sulfate and the balance being yellow serofluid, wherein the pH value is adjusted to 7.0-7.5. According to the method for producing the anti-microbial peptide, the cell number of bacillus subtilis in obtained fermentation broth is large, and the produced anti-microbial peptide is high in titer.

Description

technical field [0001] The invention relates to the field of microbial fermentation, in particular to a subtilis culture medium for producing antimicrobial peptides and its application. Background technique [0002] Bacillus subtilis is a spore-forming Gram-positive bacterium that can produce antibacterial peptides such as subtilisin, polymyxin, and gramicidin during its growth, which can significantly inhibit intestinal or plant pathogenic bacteria It exerts a probiotic effect by reducing free oxygen in the intestinal environment, promoting the growth of probiotic microorganisms such as lactic acid bacteria, and regulating intestinal immunity. Based on the broad antibacterial spectrum and high stability of the antimicrobial peptides produced by Bacillus subtilis, the antimicrobial peptides produced by Bacillus subtilis have been widely used in the research of feed additives, and good application results have been achieved. Because microbial production of antimicrobial pept...

Claims

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Application Information

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IPC IPC(8): C12P21/00C12R1/125
Inventor 周辉方俊田云葛龙
Owner HUNAN AGRICULTURAL UNIV
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