De novo design, non-chromatographic purification and salt-effect of elastin-like polypeptides.
- Author:
Kaizong HUANG
1
;
Jingjing LI
;
Wei LI
;
Huihua GE
;
Wenyan WANG
;
Guangya ZHANG
Author Information
1. Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen 362021, China.
- Publication Type:Journal Article
- MeSH:
Chromatography;
Elastin;
biosynthesis;
genetics;
isolation & purification;
Escherichia coli;
genetics;
metabolism;
Genetic Vectors;
genetics;
Peptides;
genetics;
isolation & purification;
metabolism;
Recombinant Proteins;
biosynthesis;
genetics;
isolation & purification;
Sodium Chloride;
pharmacology
- From:
Chinese Journal of Biotechnology
2011;27(4):653-658
- CountryChina
- Language:Chinese
-
Abstract:
Elastin-like polypeptides (ELPs) are temperature sensitive biopolymers composed of a Val-Pro-Gly-Xaa-Gly pentapeptide repeat that derived from a structural motif found in mammalian elastin. It was a promising tag for recombinant protein purification. Here, we de novo designed a novel ELPs gene and cloned it into the modified expression vector pET-22b(+). Then, we transformed the recombinant expression vector pET-22b-ELPs into Escherichia coli BL21(DE3). Upon induction by Isopropyl beta-D-Thiogalactoside (IPTG), ELPs was expressed and purified by a non-chromatographic purification method named inverse temperature cycling. The influences of salts types and concentrations on ELPs were also determined. The results showed that the transition temperature of the [KV8F-20] decreased to 19 degrees C by 0.4 mmol/L Na2CO3. Due to its small molecular weight and sensitivity to salt, the ELPs might be a useful purification tag, which can provide a reliable and simple non-chromatographic method for purification of the recombinant protein by inverse transition cycling.
