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Characterization of a novel low molecular weight sucrase from filamentous fungus Termitomyces clypeatus

Paper ID Volume ID Publish Year Pages File Format Full-Text
35057 45071 2009 8 PDF Available
Title
Characterization of a novel low molecular weight sucrase from filamentous fungus Termitomyces clypeatus
Abstract

An extracellular sucrase from the culture filtrate of filamentous basidiomycota Termitomyces clypeatus grown on high sucrose (5%, w/v) was purified by gel filtration chromatography, ion exchange chromatography and HPGPLC. The biochemical properties, molecular weight and conformation of sucrase produced were significantly different from the sucrase earlier purified from sucrose (1%, w/v) medium in the fungus. Purified sucrase was characterized as a low molecular weight protein of 13.5 kDa as approximated by SDS-PAGE and HPGPLC and exhibited predominantly random coil conformation in far-UV CD spectra. The enzyme was optimally active at 47 °C and pH 5.0. Km and catalytic activity of the enzyme for sucrose were found to be 3.5 mM and 1.06 U/mg/mM, respectively. The enzyme was maximally active towards sucrose than to raffinose and sucrase activity was significantly inhibited by bivalent metal ions and reducing group agents. The results indicated that due to changes in aggregation pattern, molecular organization of purified sucrase, produced in high sucrose medium, was altered and was different from the previously reported enzyme. This is the first report of a sucrase of such low size showing activity.

Keywords
S5, sucrase produced in 5% sucrose medium; S1, sucrase produced in 1% sucrose medium; C, cellobiase; CF, culture filtrate; S/C, sucrase to cellobiase activity ratio; GOD-POD, glucose oxidase-peroxidase enzyme; IEC, ion exchange chromatography; HPGPLC, hig
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Characterization of a novel low molecular weight sucrase from filamentous fungus Termitomyces clypeatus
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 44, Issue 10, October 2009, Pages 1075–1082
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