Construction of novel thioredoxin fusion protein expression system and the production of recombinant Lf-CATH2.
- Author:
Yiling LU
;
Jiuxiang GAO
;
Xue QIAO
;
Yipeng WANG
;
Haining YU
- Publication Type:Journal Article
- MeSH:
Cathelicidins;
biosynthesis;
Chromatography, Affinity;
Escherichia coli;
Genetic Vectors;
Recombinant Fusion Proteins;
biosynthesis;
Thioredoxins;
genetics
- From:
Chinese Journal of Biotechnology
2015;31(3):403-410
- CountryChina
- Language:Chinese
-
Abstract:
The objective of this study was to construct an improved thioredoxin fusion protein expression system, and express the cathelicidin-derived peptide, Lf-CATH2. The improved fusion vector Lf-CATH2-pET32α(-TS) was successfully constructed by firstly deleting the thrombin site and S tag from the pET-32α vector, then inserting the Lf-CATH2 plus a thrombin site instead. Afterwards, Lf-CATH2 was expressed in Escherichia coli as fusion protein. After the cleavage by thrombin, Lf-CATH2 was released and subsequently separated using affinity chromatography. The antimicrobial activity of purified Lf-CATH2 was also examined. The improved expression vector significantly increased enzyme cleavage efficiency by 37%, and Lf-CATH2 could be expressed in high yield and maintain the biological activity. This novel thioredoxin fusion protein expression system enables a quick production of high-yield bioactive cationic peptides like cathelicidins.
