Production and characterization of a recombinant beta-1,4-endoglucanase (glycohydrolase family 9) from the termite Reticulitermes Flavipes

Xuguo Zhou, Elena S. Kovaleva, Dancia Wu-Scharf, James H. Campbell, George W. Buchman, Dion G. Boucias, Michael E. Scharf

Research output: Contribution to journalArticlepeer-review

31 Scopus citations

Abstract

Cell-1 is a host-derived beta-1,4-endoglucanase (Glycohydrolase Family 9 [GHF9]) from the lower termite Reticulitermes flavipes. Here, we report on the heterologous production of Cell-1 using eukaryotic (Baculovirus Expression Vector System; BEVS) and prokaryotic (E. coli) expression systems. The BEVS-expressed enzyme was more readily obtained in solubilized form and more active than the E. coli-expressed enzyme. Km and Vmax values for BEVS-expressed Cell-1 against the model substrate CMC were 0.993% w/v and 1.056 μmol/min/mg. Additional characterization studies on the BEVS-expressed enzyme revealed that it possesses activity comparable to the native enzyme, is optimally active around pH 6.5-7.5 and 50-60°C, is inhibited by EDTA, and displays enhanced activity up to 70°C in the presence of CaCl2. These findings provide a foundation on which to begin subsequent investigations of collaborative digestion by coevolved host and symbiont digestive enzymes from R. flavipes that include GHF7 exoglucanases, GHF1 beta glucosidases, phenol-oxidizing laccases, and others.

Original languageEnglish
Pages (from-to)147-162
Number of pages16
JournalArchives of Insect Biochemistry and Physiology
Volume74
Issue number3
DOIs
StatePublished - Jul 2010

Keywords

  • Baculovirus
  • Bioethanol
  • Endoglucanase
  • Feedstock
  • GHF9
  • Termite

ASJC Scopus subject areas

  • Physiology
  • Biochemistry
  • Insect Science

Fingerprint

Dive into the research topics of 'Production and characterization of a recombinant beta-1,4-endoglucanase (glycohydrolase family 9) from the termite Reticulitermes Flavipes'. Together they form a unique fingerprint.

Cite this