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Cloning and characterization of a putative β-glucosidase (NfBGL595) from Neosartorya fischeri

Paper ID Volume ID Publish Year Pages File Format Full-Text
34683 45039 2012 7 PDF Available
Title
Cloning and characterization of a putative β-glucosidase (NfBGL595) from Neosartorya fischeri
Abstract

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 1590 bp, 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 1693 U mg-protein−1 and a Km of 2.8 mM 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.

► We characterize a β-glucosidase (NfBGL595) from Neosartorya fischeri. ► The recombinant BGL shows high levels of catalytic activity for p-nitrophenyl-β-d-glucopyranoside. ► Molecular dynamics simulation studies shed light on the substrate specificity of NfBGL595 only towards aryl glycoside. ► NfBGL595 serves as a potential source for use in industrial application such as wine processing.

Keywords
Characterization; β-Glucosidase; Glycoside hydrolase family 1; Molecular dynamics simulation; Wine aroma
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Cloning and characterization of a putative β-glucosidase (NfBGL595) from Neosartorya fischeri
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 47, Issue 1, January 2012, Pages 99–105
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us