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Method for increasing yield of transglutaminase produced by fermentation

A technology of transglutaminase and production, which is applied in the field of bioengineering and can solve problems such as disadvantages

Active Publication Date: 2021-04-23
EAST CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This kind of autolysis is extremely unfavorable to both bacterial metabolism and product synthesis.

Method used

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  • Method for increasing yield of transglutaminase produced by fermentation
  • Method for increasing yield of transglutaminase produced by fermentation
  • Method for increasing yield of transglutaminase produced by fermentation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] 0, 0.33, 1, 3, 6, 9 g / L soybean polysaccharides were added to the fermentation medium, and samples were taken at 18, 23, 28, 36, and 47 hours after inoculation and fermentation to determine the MTG enzyme activity in the supernatant of the fermentation broth.

[0042] MTG results such as figure 1 shown. At 23 hours of fermentation, the MTG levels in the groups added with 3, 6, and 9 g / L soybean polysaccharides were significantly higher than the highest levels at 36 hours in the control group (soybean polysaccharide concentration was 0) (P values ​​were 0.0175, 0.0003, and 0.0010, respectively) When fermented to 28h, there was no significant difference in the MTG level of adding 0.33g / L soybean polysaccharide group and matched group, while adding MTG level in the 1g / L soybean polysaccharide group was significantly higher than the highest level when fermented to 36h in the control group ( P value is 0.0071). That is, the MTG enzyme activity of the MTG level in the soybe...

Embodiment 2

[0044] 0 and 3 g / L soybean polysaccharides were respectively added to the fermentation medium for inoculation and fermentation. The MTG enzyme activity in the supernatant of the 3-54h fermentation broth was measured. MTG enzyme activity as figure 2 shown. Soybean polysaccharide (3g / L) increased the maximum enzyme activity of MTG by 11%, and the maximum enzyme production time was advanced from 48h to before 30h.

Embodiment 3

[0046] 0 and 6 g / L soybean polysaccharides were respectively added to the fermentation medium for inoculation and fermentation. Samples were taken every 6 hours from 12h to 48h, and the MTG enzyme activity in the supernatant of the fermentation broth was measured and analyzed by SDS-PAGE gel electrophoresis. MTG enzyme activity is shown in Table 1. The addition of 6g / L soybean polysaccharides advanced the production time of MTG and increased the maximum enzyme activity of MTG by 10.7% (0g / L, 48h; 3g / L, 42h).

[0047] SDS-PAGE electrophoresis results such as image 3 shown. Soybean polysaccharides affected the shearing time of MTG precursor (pro-MTG) in the culture medium. In the control culture medium, a small amount of pro-MTG still remained in the fermentation broth until 42h, while adding 6g / L soybean polysaccharides fermented to At 24h, the residual amount of pro-MTG was very small, and at 30h, it could be completely secreted and matured by shearing immediately. This f...

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Abstract

The invention discloses a method for increasing yield of transglutaminase produced by fermentation. According to the method, Streptoverticillium mobaraense DSM 40587 is adopted as an experimental strain, slant spores are subjected to seed culture and then inoculated into a fermentation medium added with soybean polysaccharide to be fermented to prepare transglutaminase (MTG), and results show that expression level of the transglutaminase is remarkably increased. Wherein addition of 6 g / L of soybean polysaccharide shows the best effect. The method has advantages that the soybean polysaccharide used is non-toxic, harmless, low in cost, convenient to operate, and can improve efficiency of industrial production of transglutaminase and reduce economic costs. Under the same experimental conditions, the addition of soybean polysaccharides can advance enzyme production time by 27.6% or more, significantly shortening fermentation time.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and relates to a method for increasing the yield of transglutaminase produced by fermentation, in particular to a method for realizing high-efficiency expression of transglutaminase from Streptomyces verticillium mobara by using soybean polysaccharide as a fermentation additive . Background technique [0002] Transglutaminase (Transglutaminase, EC2.3.2.13 referred to as TGase) is widely distributed in various organisms and plays an important physiological role in the organism. TGase can catalyze the amido transfer reaction between the γ-amide group of glutamine residues in proteins and the ε-amino group of lysine to form a heterotype peptide bond of ε-(γ-glutamine)-lysine, which can promote Protein intramolecular cross-linking, protein intermolecular cross-linking, and protein-amino acid cross-linking are known as "super adhesives in the 21st century". Therefore, it has broad application...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12N1/20C12R1/465
CPCC12N9/1044C12N1/20C12Y203/02013
Inventor 高红亮金明飞刘颖牛延宁蒋德明常忠义
Owner EAST CHINA NORMAL UNIVERSITY
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