Geobacillus strain and application thereof

A geobacillus and strain technology, applied in the field of microorganisms, can solve problems such as limited research and achieve remarkable results

Inactive Publication Date: 2016-12-21
KUNMING UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, there is very limited research on thermophilic bacteria with keratin-degrading ability, which ar

Method used

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  • Geobacillus strain and application thereof
  • Geobacillus strain and application thereof
  • Geobacillus strain and application thereof

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Experimental program
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Effect test

Embodiment 1

[0032] Embodiment 1: the acquisition of Geobacillus bacterial strain Geobacillus sp.YM6

[0033] ① The sample was collected from a 65°C neutral hot spring in Eryuan, Dali, Yunnan. Take an appropriate amount of water sample and let it stand for ten minutes to absorb 1mL of the supernatant for gradient dilution to 10 -7 , take 100 μl from each gradient and spread it on the primary screening medium; culture it at 55°C for 24 hours, and select the plate with colonies;

[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 feathers, preferably high-efficienc...

Embodiment 5

[0038] The total DNA of the strain was extracted, the target fragment was amplified with bacterial 16SrDNA universal primers, and the fragment was recovered, cloned, and sequenced; the sequence of the bacterial 16SrDNA gene fragment was compared with 16S rDNA using BLAST of NCBI, and the results showed that it was similar to that of Geobacillus sp. The homology of YMTC 1049 is 99%, and it was preliminarily identified as Geobacillus, named YM6.

Embodiment 2

[0039] Embodiment 2: Geobacillus sp.YM6 bacterial strain is to the weight loss rate determination 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 Geobacillus sp.YM6 bacteria solution (OD 6000.4) inoculated into the re-screening medium, cultured with shaking at 55°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) The calculation shows that the feather weight loss rate is 65.7%.

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Abstract

The invention discloses a geobacillus strain (Geobacillus sp. YM6) with a collection number of CGMCC NO.12089 at the China General Microbiological Culture Collection center. By using leather meal as a unique carbon-nitrogen source, the strain can be connected into a fermentation culture medium with complete leather, and the leather and pinnule are mostly degraded after 48h of fermentation culture, so that the strain has remarkable leather keratin degrading effect. Optimal acting temperature of keratinase generated by the strain is 65-70 DEG C, the keratinase is high-temperature neutral keratinase, optimal pH is 7.0, and the grain can be applied in the related production field of leather degradation.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a thermophilic Geobacillus strain and its application in degrading feathers. 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...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23K10/26C12R1/10
CPCC12N1/205C12R2001/10
Inventor 林连兵叶俊炜邓先余魏云林张琦季秀玲
Owner KUNMING UNIV OF SCI & TECH
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