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Brevibacillus brevis strain and application thereof

A technology of Bacillus brevis and strains, applied in the field of microorganisms, can solve the problems of corrosion of equipment, low efficiency of alkaline method, low yield of compound amino acids, etc., and achieve remarkable results

Inactive Publication Date: 2016-06-22
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the efficiency of the alkali method is not high, and the products are mainly polypeptides; the acid method will corrode equipment and pollute the environment, and the yield of compound amino acids is not high

Method used

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  • Brevibacillus brevis strain and application thereof
  • Brevibacillus brevis strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Example 1: Obtaining of Bacillus brevius strain Brevibacillus brevisA1

[0033] ①Collect feathers from the place where the feathers are piled up, take 5g of feathers and put them into a triangular flask filled with 100mL of sterile water, shake and mix, let stand for 10 minutes, absorb 1mL of supernatant and make gradient dilution to 10 -7 , take 100 μL of each gradient and smear it on the primary screening medium plate; incubate at 37°C for 72 hours, and pick the colonies grown on the plate;

[0034] The composition of the primary screening medium is: potassium dihydrogen phosphate 1.5g, magnesium sulfate heptahydrate 0.025g, calcium chloride 0.025g, ferrous sulfate heptahydrate 0.015g, feather meal 10g, agar 15g, distilled water 1000ml.

[0035] ② Inoculate the obtained single colonies into the re-screening medium respectively, shake culture at 37°C, observe the degradation of feathers after 48 hours, and measure the soluble protein content and the degradation rate of...

Embodiment 6

[0038] The total DNA of the strain was extracted, the target fragment was amplified with bacterial 16SrDNA universal primers, and the fragments were recovered, cloned and sequenced. The 16SrDNA similarity comparison of the bacterial 16SrDNA gene fragment sequence was performed using NCBI's BLAST, and the results showed that it was 99% homologous to Brevibacillus brevisNBRC100599. It was initially identified as Bacillus brevius and named A1.

Embodiment 2

[0039] Embodiment 2: Brevibacillus brevisA1 strain measures the weight loss rate of feather degradation

[0040] (1) Wash, dry and weigh the chicken feathers;

[0041] (2) Add 2g of chicken feathers into 100mL of re-screened medium and sterilize;

[0042](3) 1mL Brevibacillus brevisA1 bacteria solution (OD 600 0.4) inoculated into the re-screening medium, cultured with shaking at 37°C for 48 hours, observed the degradation of feathers, and the degradation effect was as follows: figure 1 shown;

[0043] (4) Filter the fermentation broth, collect the residue and dry it to a constant weight, and calculate the feather weight loss rate; the feather weight loss rate calculation formula is:;

[0044] (5) Calculations show that the feather weight loss rate is 61.5%.

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Abstract

The invention discloses a brevibacillus brevis strain Brevibacillus brevis A1, and the conservation serial number in the general microbiology center of the China committee for culture collection of microorganisms is CGMCC NO.11148. Feather meal is used as a unique carbon and nitrogen source of the brevibacillus brevis strain, a fermentation medium with a complete feather is inoculated with the strain, after the strain is fermentally cultured for 48 h, most of the feather and barbule is degraded, and the effect of degrading feather keratin is remarkable; the optimum action temperature of keratinase generated by the strain is 55-60 DEG C, keratinase is medium-temperature andneutral, the optimum pH value is 7.0, and the brevibacillus brevis strain can be applied to related production field of feather degradation.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a bacillus brevis strain and its application in degrading keratin. Background technique [0002] There is a large amount of discarded feathers in people's living environment, and millions of feathers are not properly utilized every year. Feathers are rich in keratin, if properly utilized, keratin can be transformed into soluble protein that can be used. However, due to the highly cross-linked disulfide bonds and hydrogen bonds, common hydrolytic enzymes such as pepsin and trypsin are difficult to degrade. [0003] The principle of the traditional physical degradation method is to use special temperature and pressure conditions to hydrolyze feathers into soluble polypeptides or oligopeptide mixtures, and high-pressure steam can increase the solubility of keratin. The process of this method is simple. The feathers are removed from impurities, put into the react...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12R1/01
CPCC12R2001/01C12N1/205
Inventor 林连兵周少威邓先余魏云林张琦季秀玲
Owner KUNMING UNIV OF SCI & TECH
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