Heterologous expression and characterization of endocellulase from the J apanese pine sawyer M onochamus saltuarius ( M s GHF5 )

• Published in: Entomological research 2015-01, Vol.45 (1), p.26-30
• Main Authors: Ko, Hyunjun, Kwon, Hyuk‐Min, Ko, Hyun‐Jin, Ryu, Ji‐Hyeon, Song, Hwi‐Seung, Park, Yong Chul
• Format: Article
• Language: English
• ISSN: 1738-2297; 1748-5967
• DOI: 10.1111/1748-5967.12089

In this study, the endocellulase gene from M onochamus saltuarius ( M s GHF5 ) was transformed into E scherichia coli ( R osetta B lue (DE3) p L ys S strain), and induced by IPTG . The molecular weight of recombinant M s GHF5 (r M s GHF5 ) was 78 k D a and was expressed as a fusion protein with maltose binding protein in p MAL ‐c2 expression vector. Native‐ PAGE was conducted with 0.1% carboxymethyl cellulose as a substrate, and the zymogenic bands were observed. The M ichaelis constant and maximum velocity of r M s GHF5 were 0.199 mg/m L and 0.034 μmol/min/m L , respectively. The optimal condition for r M s GHF5 occurred at p H  5 and 30°C. Fe 2+ and Mn 2+ stimulated the activity of r M s GHF5 by 167 and 114% respectively, whereas Cu 2+ , Hg 2+ and Zn 2+ inhibited its activity.