High-Yield Soluble Expression and Simple Purification of the Antimicrobial Peptide OG2 Using the Intein System in Escherichia coli
Joint Authors
Xie, Yong-Gang
Han, Fei-Fei
Luan, Chao
Zhang, Hai-Wen
Feng, Jie
Choi, Young-Jun
Groleau, Denis
Wang, Yi-Zhen
Source
Issue
Vol. 2013, Issue 2013 (31 Dec. 2013), pp.1-6, 6 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2013-07-02
Country of Publication
Egypt
No. of Pages
6
Main Subjects
Abstract EN
OG2 is a modified antimicrobial peptide, that is, derived from the frog peptide Palustrin-OG1.
It has high antimicrobial activity and low cytotoxicity, and it is therefore promising as a therapeutic agent.
Both prokaryotic (Escherichia coli) and eukaryotic (Pichia pastoris) production host systems were used to produce OG2 in our previous study; however, it was difficult to achieve high expression yields and efficient purification.
In this study, we achieved high-yield OG2 expression using the intein fusion system.
The optimized OG2 gene was cloned into the pTWIN1 vector to generate pTWIN-OG2-intein2 (C-terminal fusion vector) and pTWIN-intein1-OG2 (N-terminal fusion vector).
Nearly 70% of the expressed OG2-intein2 was soluble after the IPTG concentration and induction temperature were decreased, whereas only 42% of the expressed of intein1-OG2 was soluble.
Up to 75 mg of OG2-intein2 was obtained from a 1 l culture, and 85% of the protein was cleaved by 100 mM DTT.
Intein1-OG2 was less amenable to cleavage due to the inhibition of cleavage by the N-terminal amino acid of OG2.
The purified OG2 exhibited strong antimicrobial activity against E.
coli K88.
The intein system is the best currently available system for the cost-effective production of OG2.
American Psychological Association (APA)
Xie, Yong-Gang& Han, Fei-Fei& Luan, Chao& Zhang, Hai-Wen& Feng, Jie& Choi, Young-Jun…[et al.]. 2013. High-Yield Soluble Expression and Simple Purification of the Antimicrobial Peptide OG2 Using the Intein System in Escherichia coli. BioMed Research International،Vol. 2013, no. 2013, pp.1-6.
https://search.emarefa.net/detail/BIM-1005042
Modern Language Association (MLA)
Xie, Yong-Gang…[et al.]. High-Yield Soluble Expression and Simple Purification of the Antimicrobial Peptide OG2 Using the Intein System in Escherichia coli. BioMed Research International No. 2013 (2013), pp.1-6.
https://search.emarefa.net/detail/BIM-1005042
American Medical Association (AMA)
Xie, Yong-Gang& Han, Fei-Fei& Luan, Chao& Zhang, Hai-Wen& Feng, Jie& Choi, Young-Jun…[et al.]. High-Yield Soluble Expression and Simple Purification of the Antimicrobial Peptide OG2 Using the Intein System in Escherichia coli. BioMed Research International. 2013. Vol. 2013, no. 2013, pp.1-6.
https://search.emarefa.net/detail/BIM-1005042
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1005042