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Polypeptide for inhibiting activity of bacillus subtilis transglutaminase (BTG) and its screening method and use

A technology of transglutaminase and Bacillus subtilis, applied in the biological field, can solve problems such as toxicity, low enzyme yield, and no patent publications have been found, and achieve high expression, release inhibition effect, and reduce the effect of toxic effect.

Active Publication Date: 2014-05-28
TIANJIN UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because TG has protein cross-linking activity and has toxic effects on host cells, although Portuguese Placido et al. realized the expression of BTG in E. coli in 2007, the enzyme yield was very low under different induction and culture conditions
Currently, it is still difficult to prepare large quantities of Bacillus subtilis-derived BTG
[0004] Through searching, no patent publication related to the patent application of the present invention has been found:

Method used

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  • Polypeptide for inhibiting activity of bacillus subtilis transglutaminase (BTG) and its screening method and use
  • Polypeptide for inhibiting activity of bacillus subtilis transglutaminase (BTG) and its screening method and use
  • Polypeptide for inhibiting activity of bacillus subtilis transglutaminase (BTG) and its screening method and use

Examples

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Embodiment Construction

[0029] The present invention will be described in further detail below through specific examples. The following examples are only descriptive and not restrictive, and the protection scope of the present invention cannot be limited by this.

[0030] The methods in the following examples are conventional methods unless otherwise specified.

[0031] Streptomyces-derived TG has a signal peptide and a propeptide. Due to the action of the propeptide, the Streptomyces-derived TG is inactive in the cell and cannot function as a cross-linking protein, thereby not damaging the cell. However, BTG derived from Bacillus subtilis has no signal peptide and propeptide, only mature peptides with activity. Therefore, it is hoped that the propeptide of Streptomyces TG can inhibit the activity of BTG.

[0032] The TG propeptide sequences derived from 7 different Streptomyces (S.cinnamoneus, S.caniferus, S.fradiae, S.hygroscopicus, S.mobaraense, S.netropsis, S.platensis) were selected and compared, and t...

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Abstract

The invention relates to a polypeptide for inhibiting an activity of bacillus subtilis transglutaminase (BTG) and its screening method and use. The polypeptide has an amino acid sequence shown in the formula of SEQ ID No: 1. The polypeptide provided by the invention can inhibit the activity of BTG by a fusion protein and has an inhibition rate more than 60%. Through the polypeptide, the activity of expressed BTG having strong cytotoxicity is inhibited in a host cell and then is recovered after the inhibitor is removed under the action of a blood coagulation factor Xa.

Description

Technical field [0001] The present invention belongs to the field of biotechnology, in particular to a polypeptide that inhibits the activity of Bacillus subtilis transglutaminase (Bacillus subtilis transglutaminase for short) and its screening method and application. Background technique [0002] Transglutaminase (EC2.3.2.13, referred to as TG), is an enzyme that catalyzes the acyl transfer reaction, which can catalyze the formation of ε-(γ-Glu )-Lys isopeptide bond makes it covalently cross-linked and polymerized, thereby directly changing the structure and function of the protein itself and the cells and tissues to which the protein is attached. Therefore, the enzyme has a wide range of applications in the food industry and the pharmaceutical industry. TG production methods include animal and plant tissue extraction method and microbial fermentation method. The early production of TG used animal tissue extraction method, but due to the few animal tissue sources, complex extra...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/10C12N15/70G01N21/64C12R1/54C12R1/55C12R1/585C12R1/465
Inventor 路福平刘逸寒蔺松王春霞王建玲
Owner TIANJIN UNIV OF SCI & TECH
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