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Strain for producing high-activity keratin hydrolase and application of strain

A keratin and hydrolase technology, applied in the field of bioengineering, can solve the problems of increasing the production cost of feather meal, difficult to degrade feather protein, poor digestion and the like

Active Publication Date: 2017-02-22
SHANGHAI TRANSGENIC RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The analysis of feather protein shows that sulfur-containing amino acids account for a relatively large proportion of the total amino acids. In this way, in addition to many hydrogen bonds and salt bonds, there are also a large number of disulfide bonds in the feather protein molecule and between molecules. Therefore, Ordinary proteases are difficult to degrade feather protein, resulting in poor digestion, which is one of the main problems of feather protein as a protein source for animal feed
[0004] At present, the industrial method of processing feather meal is generally high-temperature and high-pressure cooking, which will lead to the denaturation of some heat-sensitive amino acids, the production of some non-nutritive amino acids, the decrease of product stability, and the indigestion of animals after eating.
Although the microbial degradation method has mild reaction conditions, it is still in the stage of laboratory research. On the other hand, because it consumes a lot of energy during the fermentation process, it will greatly increase the production cost of feather meal, so it cannot be used in actual production. application
[0005] Currently known bacteria that decompose keratin are distributed in bacteria, actinomycetes and fungi, mainly including Bacillus licheniformis, Thermomonas, Candida albicans and Streptomyces, etc. Ineffective

Method used

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  • Strain for producing high-activity keratin hydrolase and application of strain
  • Strain for producing high-activity keratin hydrolase and application of strain
  • Strain for producing high-activity keratin hydrolase and application of strain

Examples

Experimental program
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Effect test

preparation example Construction

[0092] Preparation and formulation of keratinase

[0093] In the present invention, the keratinase based on the strain of the present invention and the preparation containing the keratinase are also provided.

[0094] In the present invention, a preferred keratinase preparation is the crude enzyme preparation of the strain of the present invention, that is, the solution obtained after the fermentation product of the strain of the present invention is centrifuged to remove solid impurities (including bacteria). The crude enzyme solution can be used directly as an enzyme preparation, or can be further processed (such as freeze-drying) to obtain a solid-state enzyme preparation.

[0095] In the enzyme preparation of the present invention, substances beneficial to maintaining enzyme activity, such as stabilizers, etc., may optionally be added.

[0096] Method for hydrolyzing keratin (application)

[0097] The present invention also provides a method for hydrolyzing (or enzymolyz...

Embodiment 1

[0121] Screening and mutagenesis of embodiment 1 bacterial strain

[0122] 1.1. Screening and isolation of strains

[0123] Take a certain amount of keratin accumulation (0.1-1g), add it into a 250ml Erlenmeyer flask containing 50mL enriched medium, shake and culture at 37°C and 180r / min for 5-7 days, and observe the color change of the medium.

[0124] The enriched culture solution was serially diluted, and the dilution was 10 -5 、10 -6 and 10 -7 The dilutions were spread on the feather meal screening plate respectively, and the grown single colonies were picked and cultured again on the screening plate for separation and purification to obtain single colonies. The single colonies obtained above were stored in test tubes for future use.

[0125] As a result, 27 different strains were screened, and their enzyme activities were within 300U / ml.

[0126] 1.2. Enzyme activity screening

[0127] Inoculate the strain obtained and preserved in the previous step into a 250ml Erl...

Embodiment 2

[0133] The optimization of embodiment 2 fermentation conditions

[0134] In this example, the culture conditions and medium were optimized for Deinococcus radiodurans J1 obtained in Example 1.

[0135] 2.1. Effect of different initial pH on enzyme production

[0136] The initial pH was set to 5, 6, 7, 8, 9, 10, and the effect of initial pH on fermentation enzyme production was investigated.

[0137] The result is as figure 2 shown. Wherein, when the pH is neutral (eg, the pH is 8), the enzymolysis rate of the feather is the highest (up to 64%), and the growth of the bacterial strain is better, which is more favorable for fermentation and enzyme production.

[0138] 2.2. Effect of different inoculation volumes on enzyme production

[0139] The inoculum volume was designed to be 4%, 6%, 8%, 10%, and 12%, and the influence of the inoculum volume on the growth of the strain was investigated.

[0140] The result is as image 3 shown. Among them, the enzymolysis rate of feat...

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Abstract

The invention provides a production strain of high-yield keratin hydrolase and application of the strain, and particularly relates to anti-radiation deinococcus for producing keratin hydrolase. The strain expresses keratin hydrolase high in activity and wide in application range. The anti-radiation deinococcus can efficiently produce keratin hydrolase at low cost, and is very high in application value.

Description

technical field [0001] The invention belongs to the field of bioengineering. Specifically, the invention relates to a bacterial strain producing highly active keratin hydrolase and application thereof. Background technique [0002] Feather is an important by-product of poultry breeding and slaughtering industry. my country has abundant feather production. At present, the annual output of feather reaches more than 1.2 million tons. Feathers are rich in protein and amino acids. Reasonable use of feathers can reduce the pollution of waste to the environment on the one hand, and on the other hand can be used as a feed protein for livestock and poultry feeding. [0003] Feathers are the outer protective layer of birds, and they are highly resistant to stress. The analysis of feather protein shows that sulfur-containing amino acids account for a relatively large proportion of the total amino acids. In this way, in addition to many hydrogen bonds and salt bonds, there are also a l...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/52C12P21/06
Inventor 成国祥江国永蒋兴龙
Owner SHANGHAI TRANSGENIC RES CENT
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