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Biochemical characterization of an extracellular polygalacturonase from Trichoderma harzianum

Paper ID Volume ID Publish Year Pages File Format Full-Text
25559 43581 2006 11 PDF Available
Title
Biochemical characterization of an extracellular polygalacturonase from Trichoderma harzianum
Abstract

An extracellular polygalacturonase (PGII) from Trichoderma harzianum was purified to homogeneity by two chromatography steps using DEAE-Sepharose and Sephacryl S-200. The molecular weight of T. harzianum PGII was 31,000 Da by gel filtration and SDS-PAGE. PGII had isoelectric point of 4.5 and optimum pH of 5.0. PGII was very stable at the pH 5.0. The extent of hydrolysis of different pectins by enzyme was decreased with increasing of degree of esterification (DE). PGII had very low activity toward non-pectic polysaccharides. The apparent Km value and Kcat value for hydrolyzing polygalacturonic acid (PGA) were 3.4 mg/ml and 592 s−1, respectively. PGII was found to have temperature optimum at 40 °C and was approximately stable up to 30 °C for 60 min of incubation. All the examined metal cations showed inhibitory effects on the enzyme activity. A 1,10-phenanthroline, Tween 20, Tween 80, Triton X-100 and SDS had no effect on the enzyme activity. The rate of enzyme catalyzed reduction of viscosity of solutions of PGA or pectin was higher three times than the rate of release of reducing sugars indicating that the enzyme had an endo-action. The storage stability of the enzyme in liquid and powder forms was studied, where the activity of the powder form was stable up to 1 year. These properties of T. harzianum PGII with appreciable activity would be potentially novel source of enzyme for food processing.

Keywords
Polygalacturonase; Pectin; Trichoderma harzianum; Purification; Characterization; Kinetic parameters
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Biochemical characterization of an extracellular polygalacturonase from Trichoderma harzianum
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 127, Issue 1, 15 December 2006, Pages 54–64
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