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Tandem multimer expression of angiotensin I‐converting enzyme inhibitory peptide in Escherichia coli

Tandem multimer expression of angiotensin I‐converting enzyme inhibitory peptide in Escherichia coli It is common for small tandem peptide multimer genes to be indirectly inserted into expression vectors and fused with a protein tag. In this study, a multimer of the tandem angiotensin I‐converting enzyme inhibitory peptide (ACE‐IP) gene was directly transferred to a commercially available vector and the designed gene was expressed as a repeated peptide in Escherichia coli BL21(DE3)pLysS. The process further developed in our study was the construction of six‐repeated ACE‐IP synthetic genes and their direct insertion. Protein expression in inclusion bodies was confirmed by SDS‐PAGE and Western blot. Acid hydrolysis of inclusion bodies produced single‐unit peptides through cleavage of the aspartyl‐prolyl bonds. This cleaved recombinant peptide (rACE‐IP) was purified using immuno‐affinity chromatography followed by reversed phase‐HPLC. 105–115 mg of the lyophilized recombinant peptide was obtained from 1 L E. coli culture. In vitro biological activity of rACE‐IP was indistinguishable from that of the natural peptide produced by hydrolysis in artificial gastric juice or by acidic hydrolysis. The rACE‐IP prepared by recombinant DNA technology and solid‐phase synthesis methods showed a similar IC50. This strategy could be used for the expression of important peptides, which have N‐terminal proline (P) and C‐terminal aspartic acid residues (D) for commercial applications, e.g. functional foods and drinks. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biotechnology Journal Wiley

Tandem multimer expression of angiotensin I‐converting enzyme inhibitory peptide in Escherichia coli

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References (42)

Publisher
Wiley
Copyright
Copyright © 2009 Wiley Subscription Services, Inc., A Wiley Company
ISSN
1860-6768
eISSN
1860-7314
DOI
10.1002/biot.200800326
pmid
19396904
Publisher site
See Article on Publisher Site

Abstract

It is common for small tandem peptide multimer genes to be indirectly inserted into expression vectors and fused with a protein tag. In this study, a multimer of the tandem angiotensin I‐converting enzyme inhibitory peptide (ACE‐IP) gene was directly transferred to a commercially available vector and the designed gene was expressed as a repeated peptide in Escherichia coli BL21(DE3)pLysS. The process further developed in our study was the construction of six‐repeated ACE‐IP synthetic genes and their direct insertion. Protein expression in inclusion bodies was confirmed by SDS‐PAGE and Western blot. Acid hydrolysis of inclusion bodies produced single‐unit peptides through cleavage of the aspartyl‐prolyl bonds. This cleaved recombinant peptide (rACE‐IP) was purified using immuno‐affinity chromatography followed by reversed phase‐HPLC. 105–115 mg of the lyophilized recombinant peptide was obtained from 1 L E. coli culture. In vitro biological activity of rACE‐IP was indistinguishable from that of the natural peptide produced by hydrolysis in artificial gastric juice or by acidic hydrolysis. The rACE‐IP prepared by recombinant DNA technology and solid‐phase synthesis methods showed a similar IC50. This strategy could be used for the expression of important peptides, which have N‐terminal proline (P) and C‐terminal aspartic acid residues (D) for commercial applications, e.g. functional foods and drinks.

Journal

Biotechnology Journal – Wiley

Published: Sep 1, 2009

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