A bacterial strain co-producing chitosanase and γ-polyglutamic acid and its application

A technology for producing chitosanase and polyglutamic acid, applied in the direction of glycosylase, enzymes, bacteria, etc., can solve the problems that have not been reported on the co-production of chitosanase and polyglutamic acid strains, and achieve Effects of high γ-polyglutamic acid production capacity, good genetic stability, and simple medium composition

Active Publication Date: 2020-09-29
LUDONG UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] A strain of chitosanase production strain screened by the present invention has higher enzyme-producing activity, and at the same time it is found that when L-glutamic acid is added to the medium, it can simultaneously produce γ-polyglutamic acid. A report on the co-production of chitosanase and polyglutamic acid

Method used

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  • A bacterial strain co-producing chitosanase and γ-polyglutamic acid and its application
  • A bacterial strain co-producing chitosanase and γ-polyglutamic acid and its application
  • A bacterial strain co-producing chitosanase and γ-polyglutamic acid and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: Classification of Bacillus suffolus strains

[0034] 1) Characteristics of strains

[0035] On the preservation medium, the colonies are off-white, nearly round, with smooth, moist and raised surfaces, and irregular edges; observed by an optical microscope, the bacteria are straight rod-shaped, not chained, with blunt round ends, spores, and Gram-positive. Strain LZ303 can decompose sucrose, glucose and sorbose, but cannot decompose starch. The results of methyl red test, V-P test, oxidase, catalase, catalase and urease test were positive, and the test results of nitrate reduction, indole production and gelatin liquefaction were negative. It grew well in 10% NaCl medium, and the test results are shown in Table 1. Based on the above test results, the strain LZ303 was preliminarily identified as Bacillus sp. according to the "Common Bacteria System Identification Manual". The comparison results of specific physiological and biochemical characteristics are sh...

Embodiment 2

[0049] Embodiment 2 strain culture and chitosanase fermentation

[0050] 1) Strain activation: pick the strains and inoculate them on a solid slant medium: 2g / L sucrose, 5g / L yeast extract powder, 5g / L peptone, 5g / L tryptone, 5g / L NaCl, 18g / L agar , pH 7.0, sterilized by high-pressure steam at 121°C for 20 minutes; streak inoculated in a constant temperature incubator at 30°C for 20 hours;

[0051] 2) Preparation of liquid seeds: Pick one ring of activated strains and inoculate them in a 500mL Erlenmeyer flask containing 50mL of seed medium. The components of liquid seed medium: 3g / L sucrose, 5g / L yeast extract powder, 5g / L peptone, Tryptone 3g / L, KH 2 PO 4 1g / L, NaCl 2.5g / L, pH 7.0, sterilized by high-pressure steam at 115°C for 20min; seal with eight layers of gauze, and shake at 30°C and 200r / min for 18h to obtain liquid seeds;

[0052] 3) liquid fermentation: according to 5% inoculation amount, the components are glucose 10g / L, colloidal chitosan 5g / L, peptone 10g / L, y...

Embodiment 3

[0053] Example 3 strain culture and gamma-polyglutamic acid fermentation

[0054] 1) Strain activation: pick the strains and inoculate them on a solid slant medium: 2g / L sucrose, 5g / L yeast extract powder, 5g / L peptone, 5g / L tryptone, 5g / L NaCl, 18g / L agar , pH 7.0, sterilized by high-pressure steam at 121°C for 20 minutes; streak inoculated in a constant temperature incubator at 30°C for 18 hours;

[0055] 2) Preparation of liquid seeds: Pick one ring of activated strains and inoculate them in a 500mL Erlenmeyer flask containing 50mL of seed medium. The components of the liquid seed medium: 3g / L sucrose, 5g / L yeast extract powder, 5g / L peptone, Tryptone 5g / L, KH 2 PO 4 1g / L, NaCl 2g / L, pH 7.0, sterilized by high-pressure steam at 115°C for 20min; seal with eight layers of gauze, and shake at 30°C and 200r / min for 18h to obtain liquid seeds;

[0056] 3) liquid fermentation: according to 10% inoculum size, the composition is 30g / L of sucrose, 5g / L of glycerol, 30g / L of L-glut...

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Abstract

The invention relates to a bacterial strain producing chitosanase and polyglutamic acid and its application, which belongs to the field of biotechnology. The bacterial strain is Bacillus suffoloidum LZ303, which is isolated from seabed sediments in Yantai sea area, and is classified and named as Bacillus suffoloidum Bacillus was preserved in the China General Microorganism Culture Collection Management Center on December 21, 2017, and the preservation number is CGMCC No.15111. The bacterial strain of the present invention has the dual synthesis ability of chitosanase and γ-polyglutamic acid, and can produce chitosanase or γ-polyglutamic acid independently by controlling the medium and culture conditions, and can also co-produce chitosanase and gamma-polyglutamic acid. Moreover, the bacterial strain of the present invention has good genetic stability, simple medium components and low raw material cost, and the chitosanase produced has high enzyme activity, can be secreted extracellularly, is simple and convenient to separate and extract, has strong substrate specificity, and can be directly used for transformation production Oligochitosan has a high ability to produce γ-polyglutamic acid at the same time, and has advantages in industrial applications.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a bacterial strain co-producing chitosanase and gamma-polyglutamic acid, and a method for preparing chitosanase and gamma-polyglutamic acid by using the bacterial strain. Background technique [0002] Low-molecular-weight chitosan has a wide range of applications in biomedicine, food, and chemical industries. It has a variety of potential biological activities such as antibacterial and inducing plants to produce phytoantibiotics. It also has anti-cholesterol synthesis in the body and anti-tumor through immune stimulation. Such activities have thus attracted more and more attention. Chitosanase (chitosanase), also known as chitosan-N-acetyl-glucosaminidase, specifically hydrolyzes the β-1,4-glucosidic bonds in chitosan, and is an ideal enzyme preparation for the preparation of chitosan oligosaccharides . In addition to preparing chitooligosaccharide shell polycanase, it also has oth...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N9/42C12P13/02C12R1/07
CPCC12N1/20C12N9/2434C12P13/02C12Y302/01132C12N1/205C12R2001/07
Inventor 张兴晓张建龙
Owner LUDONG UNIVERSITY
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