Unlock instant, AI-driven research and patent intelligence for your innovation.

Improved porcine VIP (vasoactive Intestinal peptide) and method for expressing porcine VIP in escherichia coli

A technology of Escherichia coli and expression vector, which is applied in the field of porcine VIP improved peptide and its expression in Escherichia coli, can solve the problems of complex production process, insufficient antibacterial activity, and low antibacterial spectrum, and achieve broad antibacterial spectrum, antibacterial The effect of strong bacterial ability and simple production process

Inactive Publication Date: 2015-11-04
NORTHWESTERN POLYTECHNICAL UNIV
View PDF1 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide an improved pig VIP peptide and its high-efficiency expression method in Escherichia coli to solve the problems of insufficient antibacterial activity and wide antibacterial spectrum of the existing peptide, as well as the complicated process and high production cost of the existing production technology

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Improved porcine VIP (vasoactive Intestinal peptide) and method for expressing porcine VIP in escherichia coli
  • Improved porcine VIP (vasoactive Intestinal peptide) and method for expressing porcine VIP in escherichia coli
  • Improved porcine VIP (vasoactive Intestinal peptide) and method for expressing porcine VIP in escherichia coli

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0073] Antibacterial activity detection and safety evaluation method of porcine VIP improved peptide:

[0074] Natural animal-derived antimicrobial peptide is a kind of neuroimmunomodulatory peptide, which has a simple and clear structure, has a variety of biological activities, and has a small concentration, and has potential for development; however, its antibacterial activity is relatively low. In view of this, through molecular improvement, it aims to While improving its antibacterial activity, ensure its non-toxic side effects and strong stability.

[0075] Firstly, through the minimum inhibitory concentration experiment, porcine VIP and its improved peptide pVIP were detected m Antibacterial activity against Escherichia coli ATCC25922, Staphylococcus aureus ATCC25923 and Escherichia coli K88, based on which different concentrations of VIP and pVIP were detected m The hemolytic activity of red blood cells in SD rats and the effect on the proliferation of 3T3-L1 preadipoc...

Embodiment 2

[0077] The preparation method of porcine VIP improved peptide:

[0078] (1) Synthesis of pig VIP improved peptide gene:

[0079] The screened porcine VIP improved peptide is prepared by genetic engineering to improve its applicability.

[0080] According to the amino acid sequence, structural characteristics and physical and chemical properties of porcine VIP in GeneBank, as well as the structural basis of the antibacterial peptide exerting antibacterial activity, it was improved. Amino acid sequences before and after improvement are as follows:

[0081] Before improvement: HSDAVFTDNYTRLRKQMAVKKYLNSILN,

[0082] Improved: FTANYTRLRRQLAVRRYLAAILRRR,

[0083] In order to avoid the toxicity of the pig VIP improved peptide of the present invention to E. coli host bacteria, TrxA was fused with the improved antibacterial peptide VIP, and a stop codon TAA was added to the nucleotide sequence of the antimicrobial peptide VIP analogous peptide, and the 5' end The enterokinase recog...

Embodiment 3

[0100] Detection of antibacterial activity of recombinantly expressed porcine VIP improved peptide:

[0101] Detection of Antibacterial Activity of Recombinantly Expressed Porcine VIP Modified Peptide Using Oxford Cup Method, Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923 to OD 600 About 0.5, prepare 100mL of MHB medium containing 1.5% agar, cool the sterilized MH medium to about 50°C, add 1mL of the above bacterial solution, mix well and pour the plate. After the medium solidified, place an Oxford cup vertically on the surface of the medium, add the sample to be tested into the Oxford cup, and incubate at 37°C for 24 hours. Observe the size of the inhibition zone, the higher the antibacterial activity and concentration of the sample, the larger the inhibition zone. The test results are shown in Figure 5, which shows that the pig VIP improved peptide prepared by the method of recombinant expression in Escherichia coli used in the present invention has strong...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The present invention discloses an improved porcine VIP (vasoactive Intestinal peptide). The improved porcine VIP has the following amino acid sequence: FTANYTRLRRQLAVRRYLAAILGRR. The present invention further discloses a method for expressing the porcine VIP in escherichia coli, so that the problems that an existing peptide is lack of antibacterial activity and is not wide enough in an antibacterial spectrum, an existing production technique process is complicated, and the production cost is high are solved.

Description

technical field [0001] The invention belongs to the technical field of bioengineering and relates to an improved porcine VIP peptide and its expression method in Escherichia coli. Background technique [0002] At present, in the production of animal husbandry, the extensive use or even abuse of antibiotics has led to a series of serious problems such as the emergence of drug-resistant strains of pathogenic bacteria and even superbugs, the residue of antibiotics in livestock and poultry products, and the pollution of the environment with animal feces and urine, which have seriously affected livestock. Product quality and human health. Therefore, it is imminent to develop safe, efficient, and less likely to produce drug resistance, environmental protection, disease prevention and health care livestock and poultry drugs or additives to replace antibiotics. Antimicrobial peptides, a class of natural defense bioactive peptides that exist in animals, have a series of advantages s...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K14/575C12N15/70C12P21/02
Inventor 徐春兰乔想金高紫阳牛卫宁尚晓娅
Owner NORTHWESTERN POLYTECHNICAL UNIV