Journal article
Cloning and characterization of a putative β-glucosidase (NfBGL595) from Neosartorya fischeri
Department of Chemical Engineering, Konkuk University, Seoul 143-701, South Korea1
Institute of SK-KU Biomaterials, Konkuk University, Seoul 143-701, South Korea2
Whole genome sequence of Neosartorya fischeri NRRL181 revealed four putative GH1 β-glucosidases (BGLs). One BGL, NfBGL595 was successfully expressed and characterized. DNA sequence analysis revealed an open reading frame of 1590bp, encoding a polypeptide of 529 amino acid residues. The gene was cloned in pET28a and overexpressed in Escherichia coli.
The purified recombinant BGL showed high levels of catalytic activity, with Vmax of 1693Umg-protein−1 and a Km of 2.8mM for p-nitrophenyl-β-d-glucopyranoside (pNPG). The optimal temperature and pH for enzyme activity were 40°C and 6.0, respectively. The enzyme exhibited broad substrate specificity towards aryl glycosides including pNP-mannose, pNP-galactose, pNP-xylose, and pNP-cellobioside.
A homology model of NfBGL595 was constructed based on the X-ray crystal structure of Trichoderma reesei BGL2. Molecular dynamics simulation studies of the enzyme with the pNPG and cellobiose, shed light on the substrate specificity of N. fischeri BGL595 only towards aryl glycoside.
| Language: | English |
|---|---|
| Year: | 2011 |
| Pages: | 99-105 |
| ISSN: | 18733298 and 13595113 |
| Types: | Journal article |
| DOI: | 10.1016/j.procbio.2011.10.015 |
