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Chryseobacterium gleum for producing keratinase and separation method thereof

A technology of Chryseobacterium aureus and keratinase, applied in the field of microorganisms, can solve the problems of high cost of keratin, destruction of nutritional components, uneven hydrolysis degree, etc., and achieve high keratinase activity, improved product performance, and simple post-processing steps.

Inactive Publication Date: 2008-11-05
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Keratin cannot be hydrolyzed by general proteases (such as trypsin, pepsin, papain). The method of high temperature and high pressure treatment of keratin is costly, seriously destroys nutrients, and the degree of hydrolysis is uneven.

Method used

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  • Chryseobacterium gleum for producing keratinase and separation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] 1) Take 5ml of sewage containing feather waste, add it to 50ml enrichment medium, 30°C, 200rpm, enrich and cultivate for 72 hours; the composition of enrichment medium is: 5 grams of NaCl, beef extract 5 grams, 5 grams of peptone and 980 ml of water.

[0067] 2) Take 5ml of the above-mentioned enriched culture, add it to 50ml of liquid screening medium, cultivate under the same conditions, repeat the above screening step 3 times, take 10 μl of culture solution to dilute and apply it on the solid screening medium. The composition of the solid screening medium is: every 1000 milliliters contains 20 grams of feather meal, K 2 PO 4 4 grams, NaH 2 PO 4 2H 2 O 2 g, MgCl 2 ·6H 2 O 0.2 grams, 20 grams of agar powder and 980 milliliters of water.

[0068] 3) Pick a single colony and culture it in a liquid screening medium, compare and select a strain with higher keratinase activity. The composition of the liquid screening medium is: every 1000 milliliters contains 20 gr...

Embodiment 2

[0070] 1) Take 10ml of sewage containing wool waste, add it to 100ml of enrichment medium, 30°C, 200rpm, and enrich and cultivate for 48 hours; the ingredients of the enrichment medium are: 10 grams of NaCl, beef extract 6 grams, 6 grams of peptone and 980 ml of water.

[0071] 2) Take 10ml of the above-mentioned enriched culture, add it to 100ml liquid screening medium, cultivate under the same conditions, repeat the above screening step 5 times, take 20 μl of the culture solution to dilute and apply it on the solid screening medium. The composition of the solid screening medium is: every 1000 milliliters contains 40 grams of feather meal, K 2 PO 4 5 grams, NaH 2 PO 4 2H 2 O 2 g, MgCl 2 ·6H 2 O 0.3 g, agar powder 15 g and water 980 ml.

[0072] 3) Pick a single colony and culture it in a liquid screening medium, compare and select a strain with higher keratinase activity. The composition of the liquid screening medium is: every 1000 milliliters contains 40 grams of f...

Embodiment 3

[0074] 1) Take 5ml of waste water containing hair, add it to 100ml of enrichment medium, 25°C, 200rpm, enrich and cultivate for 72 hours; the composition of the enrichment medium is: 10 grams of NaCl and 5 grams of beef extract per 1000 ml, 5 grams of peptone and 980 ml of water.

[0075] 2) Take 10ml of the above-mentioned enriched culture, add it to 100ml of liquid screening medium, cultivate under the same conditions, repeat the above screening steps 4 times, take 30 μl of culture solution to dilute and apply it on the solid screening medium. The composition of the solid screening medium is: every 1000 milliliters contains 30 grams of feather meal, K 2 PO 4 4 grams, NaH 2 PO 4 2H 2 O 2 g, MgCl 2 ·6H 2 O 0.5 grams, 20 grams of agar powder and 980 milliliters of water.

[0076] 3) Pick a single colony and culture it in a liquid screening medium, compare and select a strain with higher keratinase activity. The composition of the liquid screening medium is: every 1000 ...

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Abstract

The invention discloses a chryseobacterium gleum of keratinase and a method for separating the chryseobacterium gleum. The Latin name of the chryseobacterium gleum is Chryseobacterium L99; the chryseobacterium gleum is stored in CGMCC with the collecting number of CGMCC No.2295. The invention discloses the morphological properties of the bacterium: the bacterium body has a rod shape, no spores are produced, the bacterium does not have motility and the Gram pigmentation shows negative. The invention discloses whole-cell fatty acid of the bacterium. The main components in weight percent are as follows: 47.59 percent of 13 methyl 14 acid, 15.72 percent of dihydroxy-13-methyl myristic acid and / or Omega-7-cis-13 methyl-15 acid, 13.91 percent of trihydroxy-15-methyl 16 acid, and 9.48 percent of Omega-9-cis-14 methyl hexadecanoic acid. The invention also discloses the whole series of a 16 S rDNA of the bacterium. The separating method comprises the following steps that: the common nutritional medium is bred in enrichment, the keratin is taken as the only SiCN source solid culture medium prescreening operation and is taken as the secondary screening of the only SiCN source solid culture medium. The method is effective and rapid and the strain has strong ability to produce enzyme.

Description

technical field [0001] The invention belongs to the technical field of microbes, and relates to a strain of keratinase-producing Chryseobacterium vicinus and an isolation method thereof. Background technique [0002] Keratin has a stable chemical structure, is insoluble in water, and is not easily biodegraded. It is a hard protein with strong resistance. In nature, keratin mainly exists in the form of animal hair, feathers, scales, hooves and horns. Keratin cannot be hydrolyzed by general proteases (such as trypsin, pepsin, papain). The method of high temperature and high pressure treatment of keratin has high cost, serious damage to nutritional components, and uneven degree of hydrolysis. Keratinase can specifically degrade keratin, and screening high-yielding strains of keratinase is of great significance for the recycling of keratin waste and environmental protection. [0003] Keratinase can gently and efficiently hydrolyze feather and other keratin wastes to produce fe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N1/02C12N9/50C12R1/01
Inventor 李永泉吕龙贤周宏闵杰
Owner ZHEJIANG UNIV
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