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A glucose-tolerant β-glucosidase from Prunus domestica seeds: Purification and characterization

Paper ID Volume ID Publish Year Pages File Format Full-Text
34687 45039 2012 6 PDF Available
Title
A glucose-tolerant β-glucosidase from Prunus domestica seeds: Purification and characterization
Abstract

A glucose-tolerant β-glucosidase was purified to homogeneity from prune (Prunus domestica) seeds by successive ammonium sulfate precipitation, hydrophobic interaction chromatography and anion-exchange chromatography. The molecular mass of the enzyme was estimated to be 61 kDa by SDS-PAGE and 54 kDa by gel permeation chromatography. The enzyme has a pI of 5.0 by isoelectric focusing and an optimum activity at pH 5.5 and 55 °C. It is stable at temperatures up to 45 °C and in a broad pH range. Its activity was completely inhibited by 5 mM of Ag+ and Hg2+. The enzyme hydrolyzed both p-nitrophenyl β-d-glucopyranoside with a Km of 3.09 mM and a Vmax of 122.1 μmol/min mg and p-nitrophenyl β-d-fucopyranoside with a Km of 1.65 mM and a Vmax of 217.6 μmol/min mg, while cellobiose was not a substrate. Glucono-δ-lactone and glucose competitively inhibited the enzyme with Ki values of 0.033 and 468 mM, respectively.

► High tolerance of the β-glucosidase I to high concentrations of glucose. ► Good stability of the enzyme at temperatures up to 45 °C and in a broad pH range. ► High catalytic activity of the enzyme toward p-nitrophenyl β-d-fucopyranoside.

Keywords
β-Glucosidase; Kinetics; Prunus domestica; Purification; Seeds; Substrate specificity
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A glucose-tolerant β-glucosidase from Prunus domestica seeds: Purification and characterization
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 47, Issue 1, January 2012, Pages 127–132
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