Recombinant bacterium for expressing glutamine transaminage and application thereof

A technology of transglutaminase and recombinant bacteria, applied in the direction of application, transferase, fungi, etc., can solve the problem of low production of transglutaminase

Inactive Publication Date: 2010-01-27
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Transglutaminase production by Lactococcus lactis and Corynebact...

Method used

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  • Recombinant bacterium for expressing glutamine transaminage and application thereof
  • Recombinant bacterium for expressing glutamine transaminage and application thereof
  • Recombinant bacterium for expressing glutamine transaminage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Preparation of yeast strain expressing transglutaminase and its functional detection

[0027] 1. Preparation of strains expressing transglutaminase

[0028] 1. Construction of recombinant expression vector containing transglutaminase precursor gene

[0029] 1) Extraction of chromosomal DNA from Streptomyces mobaraensis

[0030] Inoculate the spore suspension or slanted mycelia of Streptomyces mobara CPCC 200365 (purchased from China Medicinal Microorganism Culture Collection Management Center) stored at -70°C into 5ml TSB medium (20g / L soluble starch, 20g / L peptone, 2g / L yeast powder, 2g / L magnesium sulfate, 2g / L potassium dihydrogen phosphate, 2g / L dipotassium hydrogen phosphate), shake culture at 30°C for 48 hours. Take 3ml of bacterial liquid and centrifuge to collect mycelia, and wash twice with 500μl of STE buffer (10mM Tris-Cl, pH8.0, 0.1mM NaCl, 1mM EDTA, pH8.0). Use 500 μl STE to fully suspend the mycelium, add lysozyme to make the final concentrat...

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Abstract

The invention discloses a recombinant bacterium which is a recombinant methanol nutritional type microzyme containing a glutamine transaminage prosoma gene and a subtilisin type hydrolase gene SAM-P45. The invention also protects the recombinant bacterium as Pichia pastoris GSMTGB, the recombinant bacterium is preserved in CGMCC (address: Datun Road, Chaoyang district, Beijing city, China) on August 25th, 2009, the preserving number is CGMCC No.3249, and the enzyme activity of the glutamine transaminage reaches 0.43u/ml when the recombinant bacterium is induced and fermented for 72 hours.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to recombinant bacteria expressing glutamine transaminase and applications thereof. Background technique [0002] Transglutaminase (Transglutaminase, also known as transglutaminase, EC 2.3.2.13, referred to as TG) widely exists in the human body, animals, plants and microorganisms, and can catalyze the acyl transfer reaction between or within protein molecules. Through these reactions, polymerization and crosslinking can be induced between or within various protein molecules. Among them, transglutaminase derived from microorganisms has become a hotspot in the research and application of transglutaminase because of its Ca2+-independence, wide substrate range and relatively low production cost. [0003] Transglutaminase has a wide range of applications in food processing: ①By catalyzing the cross-linking between protein molecules, it improves the important properties of protein such as s...

Claims

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Application Information

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IPC IPC(8): C12N1/19C12N9/10A23L1/28C12R1/84A23L31/10
Inventor 李寅朱泰承游丽金
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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