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A novel ginsenoside Rb1-hydrolyzing β-d-glucosidase from Cladosporium fulvum

Paper ID Volume ID Publish Year Pages File Format Full-Text
35161 45079 2009 7 PDF Available
Title
A novel ginsenoside Rb1-hydrolyzing β-d-glucosidase from Cladosporium fulvum
Abstract

A novel β-glucosidase (G-II) was purified to homogeneity from a culture filtrate of the phytopathogenic fungus Cladosporium fulvum (syn. Fulvia fulva). G-II specifically cleaved the β-(1 → 6)-glucosidic linkage at the C-20 site of ginsenoside Rb1 to produce ginsenoside Rd, but did not hydrolyze the other β-d-glucosidic linkages in protopanaxadiol-type ginsenosides. In specificity tests, G-II was active against pNPG and disaccharides such as cellobiose and gentiobiose, but exhibited very low activities against other aryl-glycosides and methyl-α-glycosides. G-II consisted of two identical subunits with a native molecular mass of 180 kDa and a pI of 4.4. The optimal pH of G-II was pH 5.5, and the enzyme was highly stable over a range of pH 5.0–11.0. The optimal temperature was 45 °C, and the enzyme became unstable at temperatures above 40 °C. The Km and Vmax values against pNPG were 0.19 mM and 57.7 μmol/(min mg), respectively. The enzyme was inhibited by Zn2+, Cu2+ (over 50 mM) and SDS (250 mM). However, the inhibition by SDS was partially reversed by 10 mM dithiothreitol. Three oligopeptide fragments obtained after enzymatic digestion of G-II were sequenced by nanoESI-MS/MS. The amino acid sequence homology analysis showed that G-II possessed significant homology with the family 3 β-glucosidases.

Keywords
β-Glucosidase; Cladosporium fulvum (syn. Fulvia fulva); Purification; Characterization; Ginsenoside Rb1; Ginsenoside Rd
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A novel ginsenoside Rb1-hydrolyzing β-d-glucosidase from Cladosporium fulvum
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 44, Issue 6, June 2009, Pages 612–618
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