A kind of Escherichia coli strain for producing active short peptide

A technology of Escherichia coli and Escherichia coli, which is applied in the field of genetic engineering, can solve the problems of low cost, lack of high activity in vivo, and the downstream separation process of active peptide polymers needs to be further improved, so as to achieve low cost and wide application Outlook, prevention and effects of maintaining cardiovascular health

Active Publication Date: 2017-10-20
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the use of genetic engineering technology to prepare highly active short peptides has achieved a technical breakthrough, the gene design, expression efficiency and downstream separation process of active peptide multimers need to be further improved
However, there is no report on "synchronously producing ACE inhibitory peptides and antioxidant peptides using genetically engineered bacteria"
[0006] At present, there is a lack of E. coli strains that can produce active short peptides with high in vivo activity and low cost

Method used

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  • A kind of Escherichia coli strain for producing active short peptide
  • A kind of Escherichia coli strain for producing active short peptide
  • A kind of Escherichia coli strain for producing active short peptide

Examples

Experimental program
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Embodiment 1

[0031] The invention provides a bacterial strain, an Escherichia coli strain for preparing active short peptides, the bacterial strain is Escherichia coli; the preservation name is Escherichia coli; it is preserved in the General Microbiology Center (CGMCC) of China Microbiological Culture Collection Management Committee, The deposit address is No. 3, Courtyard No. 1, Beichen West Road, Chaoyang District, Beijing; date of deposit: October 24, 2014; deposit number: CGMCC No.9842.

[0032] The Escherichia coli bacterial agent prepared by the Escherichia coli described in the present invention.

[0033] Escherichia coli bacterial agent of the present invention, its active component is at least one in following (a) (b) (c):

[0034] (a) the fermentation culture of the escherichia coli described in claim 1;

[0035] (b) the sonication supernatant of the Escherichia coli cell obtained in claim 1;

[0036] (c) ultrasonic lysis precipitation of the Escherichia coli cell obtained in ...

Embodiment 2

[0073] The difference between Example 2 and Example 1 is that the engineering bacteria pET28a-ESA3 / E.coli BL21(DE3) was constructed according to the steps (1) and (2) in Example 1.

[0074] In step (1), a single colony of the strain was placed in 20 ml of LB culture solution containing 50 μg / ml Kana at a temperature of 37° C. and a stirring rate of 200 rpm for 13 hours on a shaker.

[0075] In step (3), the Escherichia coli strain was cultivated in a shake flask until the OD value of the thalline reached 0.7, and IPTG was added to a final concentration of 0.02 mM, and the temperature was 20° C. After the induction culture time was 20 h, a sample was taken for SDS-PAGE analysis, and the prepared Escherichia coli bacteria.

[0076] Such as Figure 7 It is known that after the engineered bacteria were induced and cultured for 20 hours under the condition of low temperature and low inducer concentration, a protein band with a molecular weight of about 45 kDa appeared relative to ...

Embodiment 3

[0078] The difference between embodiment 3 and embodiment 1 is:

[0079] In step (1), a single colony of the strain was placed in 20 ml of LB culture solution containing 50 μg / ml Kana at a temperature of 37° C. and a stirring rate of 200 rpm for 14 hours on a shaker.

[0080] In step (3), the Escherichia coli strain was cultured in a shake flask until the OD value of the thalline reached 0.7, and IPTG was added to a final concentration of 0.1 mM, and the temperature was 30° C. After the induction culture time was 10 h, a sample was taken for SDS-PAGE analysis, and the prepared Escherichia coli bacteria.

[0081] Preparation of recombinant small peptide mixture and identification of its activity. Take an appropriate amount of the prepared Elps-SUMO-A3, add SUMO protease according to the addition amount of 3%, and enzymatically hydrolyze at 30°C for 5h. After the enzymolysis, add sodium chloride with a final concentration of 1.5mol / l to the enzymolysis solution, bathe in water...

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Abstract

The invention discloses an Escherichia coli strain for preparing active short peptides, the strain is Escherichia coli; the preservation name is Escherichia coli; it is preserved in CGMCC, General Microorganism Center of China Microbiological Culture Preservation Management Committee, and the preservation address is Chaoyang, Beijing No. 3, No. 1 Yard, Beichen West Road, District; date of deposit: October 24, 2014; deposit number: CGMCCNo.9842. The Escherichia coli bacterial agent prepared by the Escherichia coli described in the present invention. The present invention is an Escherichia coli strain capable of producing active short peptides with high in vivo activity and low cost.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to an Escherichia coli strain for preparing active short peptides. Background technique [0002] Escherichia coli is a Gram-negative round bacillus that lives in the gut of warm-blooded animals. Most strains of E. coli are harmless, but some serotypes can cause severe food poisoning in humans. The harmless Escherichia coli is part of the normal flora of the intestinal tract, and its production of vitamin K2 is beneficial to the host and also prevents the production of other pathogenic pathogenic bacteria in the intestinal tract. Escherichia coli accounts for 0.1% of the entire intestinal flora, and the fecal-oral transmission route is an important way for this pathogen to cause disease. At the same time, Escherichia coli is a widely studied prokaryotic model organism, because it can be used as a host for recombinant DNA, and it is an important bacterium in the field of microbiol...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21A61K35/74A61P9/12C12R1/19
Inventor 饶胜其方维明高璐杨振泉徐鑫
Owner YANGZHOU UNIV
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