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Basophilic streptomyce, neutral endoglucanase produced by basophilic streptomyce and application of basophilic streptomyce

A technology of endoglucanase and alkaliphilic streptomyces, which is applied in the field of neutral glucanase and application, can solve the problems of restricting the use and development of bacteria and molds, and achieve the benefits of large-scale fermentation production, enzyme The activity is stable and the effect of inhibiting the contamination of bacteria

Inactive Publication Date: 2015-06-03
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For reasons such as these, the use and development of bacteria and molds are limited

Method used

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  • Basophilic streptomyce, neutral endoglucanase produced by basophilic streptomyce and application of basophilic streptomyce
  • Basophilic streptomyce, neutral endoglucanase produced by basophilic streptomyce and application of basophilic streptomyce
  • Basophilic streptomyce, neutral endoglucanase produced by basophilic streptomyce and application of basophilic streptomyce

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] The isolation and cultivation of embodiment 1 bacterial strain

[0049] 1. Enrichment culture

[0050] A strain producing neutral endoglucanase was screened from mangrove soil in Shenzhen. 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.

[0051] Enrichment medium: CMC-Na 8.0g / L, peptone 10g / L, NaCl 15g / L, KH 2 PO 4 1.5g / L, Na 2 HPO 4 12H 2 O 9.0g / L, MgSO 4 ·7H 2 O 2.0g / L, pH 9.0.

[0052] 2. Flat plate primary screening

[0053] 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 CMC-Na, cultured at 37°C for 48h, and the single colony with a large transparent circle was picked into the alkaline slant mediu...

Embodiment 2

[0062] The enzymatic activity of the neutral endoglucanase produced by Example 2 Streptomyces sp.H31

[0063] (1) Materials

[0064] 1. Strains

[0065] Streptomyces alkalophilus is a neutral endoglucanase-producing strain Streptomyces sp.H31 (CCTCC NO:M 2015003) screened from Shenzhen mangrove soil.

[0066] 2. Medium

[0067] Selection medium: 10g / L CMC-Na (sodium carboxymethylcellulose), 5g / L yeast powder, 5g / L NaCl, 2g / L KH 2 PO 4 , 5g / L NH 4 NO 3 , 0.3g / L MgSO 4 , 1g / L (NH 4 ) 2 SO 4 , solid medium plus agar 15-20g / L, pH 9.0; culture temperature is 37°C.

[0068] (2) Experimental method

[0069] 1. Extraction of crude enzyme solution

[0070] Put the Streptomyces sp.H31 strain into the liquid selection medium, cultivate it at 37°C and 200rpm for 48h, freeze and centrifuge the 48h culture solution at 14000rpm for 20min, and take the supernatant; The supernatant was concentrated by ultrafiltration to obtain a crude enzyme solution.

[0071] 2. Enzyme activity ...

Embodiment 3

[0095] (1) Materials

[0096] 1. Strains. Streptomyces alkalophilus is a neutral endoglucanase-producing strain Streptomyces sp.H31 (CCTCC NO:M 2015003) screened from Shenzhen mangrove soil. E.coli TOP10F' was purchased from Invitrogen;

[0097] 2. Carrier. The Escherichia coli cloning vector pMD-19T (simple) was purchased from Dalian Bao Biological Company, and the Escherichia coli expression vector pET-28a(+) (Novagen, KanR) was purchased from Novagen Company.

[0098] 3. Medium

[0099] (1) Selection medium: 10g / L CMC-Na (sodium carboxymethylcellulose), 5g / L yeast powder, 5g / L NaCl, 2g / L KH 2 PO 4 , 5g / L NH 4 NO 3 , 0.3g / L MgSO 4 , 1g / L (NH 4 ) 2 SO 4 , solid medium plus agar 15-20g / L, pH 9.0; culture temperature is 37°C.

[0100] (2) LB medium: tryptone 10g / L, yeast extract 5g / L, NaCl 10g / L, solid medium plus agar 15-20g / L, pH 9.0, autoclaved at 121°C for 20min.

[0101] 4. Main reagents. DNA polymerase and DNA molecular weight markers are products of Bao Bio...

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Abstract

The invention discloses basophilic streptomyce, neutral endoglucanase produced by basophilic streptomyce and application of basophilic streptomyce. Basophilic streptomyce is a strain which is screened from soil of mangrove in Shenzhen and is capable of producing the neutral endoglucanase. The neutral endoglucanase has excellent enzymatic characteristics, is applicable to industries of washing and papermaking, is wide in pH application range and capable of resisting heat, alkali, metal ions and surfactants and can be applied to industries of washing and papermaking. Basophilic streptomyce belongs to an alkaline-resisting extreme microorganism, can be cultured at high-alkaline conditions and is capable of effectively restraining microbial contamination and beneficial to large-scale fermentation production.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to an alkaliphilic streptomyces and the neutral glucanase produced by it and its application. Background technique [0002] Cellulose is the main component of plant fiber, accounting for about 30-50% of its dry weight. It is the most widely distributed natural carbohydrate and the most abundant and cheapest renewable resource on the earth. With the rapid increase of the world population, the rapid development of the global economy, the shortage of food and the reduction of non-renewable resources such as oil, coal and natural gas at an alarming rate, the development and utilization of renewable cellulose resources has attracted widespread attention around the world and has become A major topic of research in countries all over the world. The study of cellulase has opened up a broad way for this, especially in the past 10 years, with the rapid development of modern b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N9/42C12R1/465
CPCC12N9/2437C12Y302/01004C12N1/205C12R2001/465
Inventor 刘森林区晓阳邢苗陈伟钊
Owner SHENZHEN UNIV
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