title: [De novo design, non-chromatographic purification and salt-effect of elastin-like polypeptides].
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.
authors.affiliation: Department of Bioengineering and Biotechnology, Huaqiao University, Xiamen 362021, China.
authors.forename: Kaizong
authors.initials: K
authors.lastname: Huang
authors.forename: Jingjing
authors.initials: J
authors.lastname: Li
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Huihua
authors.initials: H
authors.lastname: Ge
authors.forename: Wenyan
authors.initials: W
authors.lastname: Wang
authors.forename: Guangya
authors.initials: G
authors.lastname: Zhang
mesh_terms: D002845:Chromatography; D004549:Elastin / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D010455:Peptides / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification*; D012965:Sodium Chloride / Q000494:pharmacology*
chemical_list: D010455:Peptides; D011994:Recombinant Proteins; D012965:Sodium Chloride; D004549:Elastin