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Basophilic streptomyces as well as alkaline xylanase produced by basophilic streptomyce and application

A technology of streptomyces alkaliphiles and xylanase, applied in the field of alkaline xylanase and application, can solve the problems of short service life, poor stability, poor direct utilization effect, etc., and achieve the purpose of inhibiting bacterial contamination and enzymatic activity Stable and conducive to the effect of large-scale fermentation production

Inactive Publication Date: 2018-07-13
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are still some problems in the direct use of xylanase in industry: poor stability, low substrate specificity, short service life and high cost.
In addition, the specificity of xylanase is low, and the effect of direct utilization is poor. Therefore, the relationship between the type and mass concentration of the substrate and the dosage of xylanase should be studied in depth, and the compatibility effect between different enzyme preparations should be explored to improve the quality of xylanase. substrate specificity

Method used

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  • Basophilic streptomyces as well as alkaline xylanase produced by basophilic streptomyce and application
  • Basophilic streptomyces as well as alkaline xylanase produced by basophilic streptomyce and application
  • Basophilic streptomyces as well as alkaline xylanase produced by basophilic streptomyce and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] The isolation and cultivation of embodiment 1 bacterial strain

[0044] 1. Enrichment culture

[0045] Alkaline xylanase-producing strains were screened from mangrove soil. Weigh 1g soil sample, add it into 100mL sterile water, shake well to make a bacterial suspension, take 1mL bacterial suspension respectively and insert it into 5mL enrichment medium that has been sterilized beforehand, enrich Set cultured for 48h.

[0046] Enrichment medium: xylan 8.0g / L, peptone 10g / L, NaCl 15g / L, KH 2 PO 4 1.5g / L, Na 2 HPO 4 12H 2 O9.0g / L, MgSO 4 ·7H 2 O 2.0g / L, pH 9.0.

[0047] 2. Flat plate primary screening

[0048] Dilute the enriched bacterial solution 10 times step by step, and take the dilution factor as 10 -3 、10 -5 、10 -7 0.5mL of the bacterial suspension was spread on the alkaline selective medium plate containing xylan, cultured at 37°C for 48h, and the single colony with a large transparent circle was picked into the alkaline slant medium, and this was rep...

Embodiment 2

[0056] Example 2 Separation and Purification of Alkaline Xylanase Produced by Streptomyces sp.WMN-3

[0057] (1) Materials

[0058] 1. Strains

[0059] Streptomyces alkalophilus is an alkaline xylanase-producing strain Streptomycessp.WMN-3 (CCTCC NO: M 2017477) screened from mangrove soil.

[0060] 2. Medium

[0061] Selection medium: xylan 8.0g / L, KNO 3 1.0g / L, MgSO 4 ·7H 2 O 0.5g / L, NaCl 15g / L, KH 2 PO 4 1.5g / L, solid medium plus agar 15-20g / L, pH9.0; culture temperature is 37°C.

[0062] 3. Main reagents

[0063] Ion-exchange chromatography media DEAE Sepharose Fast Flow and SP-sepharose Fast Flow cation-exchange chromatography media were purchased from GE. Acrylamide (acrylamide) and N,N-methylene bisacrylamide are imported from Serva and repackaged. DNA polymerase and DNA molecular weight markers are products of Baobio; protein molecular weight markers are products of Fermentas (MBI); ethylenediaminetetraacetic acid (EDTA), ethylene glycol diethyl ether diamine...

Embodiment 3

[0082] The enzymatic property research of embodiment 3 alkaline xylanase

[0083] (1) Experimental method

[0084] 1. Optimum reaction pH and pH stability

[0085] Take an appropriate amount of the pure enzyme solution of alkaline xylanase obtained in Example 2, add 1% xylan solutions with different pH values ​​respectively, and measure the enzyme activity according to a conventional method. At the same time, the pure enzyme solution was stored at different predetermined pH conditions for 30 minutes, and then the enzyme activity was measured at pH 9.0 according to the conventional method.

[0086] 2. Optimum reaction temperature and temperature stability research

[0087] Take an appropriate amount of pure enzyme liquid of alkaline xylanase obtained in Example 2 and place them in different temperature conditions to react for 20 minutes, and measure its enzyme activity. At the same time, the pure enzyme solution was placed under different temperature conditions (20°C-70°C) a...

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Abstract

The invention discloses basophilic streptomyces as well as alkaline xylanase produced by the basophilic streptomyce and application. The basophilic streptomyces is an alkaline xylanase producing strain screened from soil of a mangrove forest. The invention further discloses the alkaline xylanase produced by the basophilic streptomyce; the alkaline xylanase has a wide pH (Potential of Hydrogen) applicable range, resists heat, alkali, metal ions, a surfactant and the like and has good enzymatic characteristics that the alkaline xylanase is applicable to industries including papermaking and the like; the alkaline xylanase can be applied to the industries including the papermaking and the like. The basophilic streptomyces disclosed by the invention belongs to an alkali-resisting extreme microorganism and can be cultured under a high-alkaline condition; the basophilic streptomyces has the characteristics that bacterium pollution can be effectively inhibited and large-scale fermentation production is easy to realize.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to an alkaliphilic streptomyces and the alkaline xylanase produced by it and its application. Background technique [0002] Xylan is a hemicellulose linked by β-1,4 linkages with the unit of xylanose, which is abundant in broad-leaved trees and most annual plants. The content of hemicellulose in nature accounts for 30% of biomass, second only to cellulose. [0003] Xylanase is a general term for enzymes that catalyze the hydrolysis of xylan into xylooligosaccharides or xylooligosaccharides, including a variety of endo-xylanases and exo-xylanases, widely distributed in bacteria, fungi, yeast, actin Bacteria, rumen of ruminants, snails, crustaceans, terrestrial plant tissues and various invertebrates, among which neutral or alkaline xylanases derived from bacteria have high alkali resistance and thermal stability. Xylanase has a wide range of applications. In the pap...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/24D21C5/00C12R1/465
CPCC12N9/248C12N1/205C12R2001/465D21C5/005
Inventor 刘森林周鹏陈伟钊
Owner SHENZHEN UNIV
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