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Stenotrophomonas maltophilia for generating keratinase and application of stenotrophomonas maltophilia

A keratinase, monomonas technology, applied in applications, bacteria, microorganism-based methods, etc., can solve problems such as wool texture damage, environmental pollution, etc.

Active Publication Date: 2012-01-25
XINYI HANLING BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, chemical methods such as chlorination and hydrogen peroxide are used to treat wool scales, which pollutes the environment.
However, when the biological treatment method is applied, the single treatment with protease will cause great damage to the texture of wool.

Method used

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  • Stenotrophomonas maltophilia for generating keratinase and application of stenotrophomonas maltophilia
  • Stenotrophomonas maltophilia for generating keratinase and application of stenotrophomonas maltophilia
  • Stenotrophomonas maltophilia for generating keratinase and application of stenotrophomonas maltophilia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Embodiment 1 produces the screening method of keratinase bacterial strain

[0022] 1. The sampling site is a local poultry farm. Add the soil or sludge samples obtained from the sampling into a triangular flask filled with 20mL of sterilized water and glass beads, mix them on a shaker at 37°C, and crush them to make a bacterial suspension. liquid;

[0023] 2. Add the bacterial suspension to the primary screening medium, 37°C, 200 rpm, until the feathers are degraded, and pass for three times.

[0024] Primary screening medium: Feather 10, K 2 HPO 4 , 1.4, KH 2 PO 4 0.7, NaCl0.5, MgSO 4 ·7H 2 O 0.1, pH7.0-7.2. (g / l)

[0025] 3. Aspirate the supernatant for gradient dilution, take 10 -4 、10 -6 、10 -6 3 dilutions spread skim milk plates. Incubate upside down at 37°C for 48 hours after coating;

[0026] 4. Select the spots with larger transparent circles and plant them in the skimmed milk medium.

[0027] Six strains could be screened out from the morphologi...

Embodiment 2

[0031] The preliminary fermentation condition determination of embodiment 2 producing keratinase bacterial strain

[0032] Different temperature, pH, nitrogen source, carbon source and inorganic salt were selected to preliminarily determine the fermentation conditions of the screened bacteria. The fermentation time was set at 40h. The results are shown in Table 1.

[0033] Enzyme activity assay: draw 1mL of appropriately diluted enzyme solution, add 1mL of 0.05mol / LTtis-Hcl buffer solution (pH7.5) and 5mg of substrate (feather meal), incubate at 40°C for 60min, add 2mL of 4M TCA solution to terminate the reaction, After standing for 20min, the insoluble matter was filtered off. Centrifuge for 5min, draw 0.5mL supernatant and transfer to a new test tube, then add 0.5mL Folinol reagent and 2mL 0.5M Na 2 CO 3 After developing the color at 40°C for 20 minutes, the absorbance value was detected at 660 nm, and each increase of 0.01 in the absorbance value was defined as one enzyme...

Embodiment 3

[0039] Embodiment 3: keratinase-producing bacteria are applied to feed processing

[0040] The screened bacterial strain is inoculated in the single feather inorganic salt medium with feather content of 20g / L, after 48 hours of fermentation, the fermentation supernatant is taken to detect the free amino acid species and content (Table 2), and the fermentation results are shown in figure 2 .

[0041] Determination of preliminary fermentation conditions of table 2 keratinase-producing strains

[0042]

[0043]

[0044] After amino acid analysis, 17 kinds of amino acids can be detected, including rare amino acids that cannot be synthesized by animals themselves. Adding the fermentation product to poultry feed can replace soybeans to meet the feed used by poultry.

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Abstract

The invention discloses stenotrophomonas maltophilia for generating keratinase and an application of stenotrophomonas maltophilia. A strain for generating keratinase can efficiently degrade feathers within 24 hours, the activity of the keratinase subjected to fermentation and primary optimization can reach 150U / mL. Multiple kinds of amino acids can be produced after the degraded feathers are subjected to amino acid analysis, and the degraded feathers can replace grain crops such as soybeans and the like to be used as a main nitrogen source of feeds. Meanwhile, a fermenting enzyme solution of the stenotrophomonas maltophilia has a better treatment effect on wool. The stenotrophomonas maltophilia for generating the keratinase, screened in the invention, has better application prospects in the industries of feeds and textiles.

Description

technical field [0001] The invention relates to a keratinase-producing monocyst and its application, in particular to the screening and application of a keratinase-producing monocyst. Background technique [0002] Keratinase is an enzyme that can specifically degrade keratin, which can be produced by various microorganisms such as bacteria, actinomycetes and fungi. The common keratin is animal hair, such as cow hair, wool and human hair. Feather, as a by-product of poultry breeding and slaughtering industry, has a huge annual output and is rich in protein and amino acids. It is a potential excellent protein resource. In the traditional process of using feathers, acid-base hydrolysis is mainly used. Thermal degradation and other methods, the former has the problem of polluting the environment, the latter consumes a lot of energy, and can destroy some amino acids, reducing the nutritional value of the product. Reasonable utilization of feathers can reduce the pollution of wa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20D06M16/00C12S11/00A23K1/16C12R1/01D06M101/12A23K10/12A23K10/18A23K50/75
Inventor 陈坚刘柏宏张娟堵国成
Owner XINYI HANLING BIO ENG
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