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Secretory expression and enzymatic characterization of recombinant Agarivorans albus β-agarase in Escherichia coli

Authors :
Hyung-Min Lee
Ji-Na Kong
Kwang-Hoon Kong
Sug-Young Yoon
Source :
Preparative Biochemistry & Biotechnology. 47:1037-1042
Publication Year :
2017
Publisher :
Informa UK Limited, 2017.

Abstract

Agarase catalyzes the hydrolysis of agar, which is primarily used as a medium for microbiology, various food additives, and new biomass materials. In this study, we described the expression of the synthetic gene encoding β-agarase from Agarivorans albus (Aaβ-agarase) in Escherichia coli. The synthetic β-agarase gene was designed based on the biased codons of E. coli to optimize its expression and extracellular secretion in an active, soluble form. The synthesized agarase gene, including its signal sequence, was cloned into the pET-26 expression vector, and the pET-Aaβ-agarase plasmid was introduced into E. coli BL21-Star (DE3) cells. The E. coli transformants were cultured for high-yield secretion of recombinant Aaβ-agarase in Luria-Bertani broth containing 0.6 mM isopropyl β-D-1-thiogalactopyranoside for 9 h at 37°C. The expressed recombinant Aaβ-agarase was purified by ammonium sulfate precipitation and diethylaminoethyl-sepharose column chromatography, yielding ∼10 mg/L Aaβ-agarase. The purified recombinant Aaβ-agarase exhibited optimal activity at pH 7 and 40°C, and its activity was strongly inhibited by Cu

Details

ISSN :
15322297 and 10826068
Volume :
47
Database :
OpenAIRE
Journal :
Preparative Biochemistry & Biotechnology
Accession number :
edsair.doi.dedup.....80f047a5987127928a578e7e2ce107ca
Full Text :
https://doi.org/10.1080/10826068.2017.1373292