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Overexpression and characterization of a thermostable, pH-stable and organic solvent-tolerant Ganoderma fornicatum laccase in Pichia pastoris

Paper ID Volume ID Publish Year Pages File Format Full-Text
34908 45059 2011 6 PDF Available
Title
Overexpression and characterization of a thermostable, pH-stable and organic solvent-tolerant Ganoderma fornicatum laccase in Pichia pastoris
Abstract

The newly defined laccase gene, lac1, from the medicinal white-rot fungus Ganoderma fornicatum 0814 was cloned and expressed in Pichia pastoris. The corresponding open reading frame has 1563 nucleotides and encodes a protein of 521 amino acids, including a 21-residue secretion signal peptide. The absorption spectrum of the purified laccase, rLac1, indicated that it is a yellow laccase rather than the common blue laccase. The optimum pH of rLac1 was 2.5, 3.0 and 3.0 for ABTS, DMP and guaiacol, respectively. The kinetic parameter Km of rLac1 was 103.9, 276.7 and 1263.4 μM for ABTS, DMP and guaiacol, respectively. The optimal temperature for rLac1 was 55 °C for all of the substrates. rLac1 is stable up to 60 °C and in the pH range 2.5–10.0. Sodium azide and thioglycolic acid strongly inhibited rLac1 activity, but it was not affected by EDTA. rLac1 retained more than 80% activity in 50% (v/v) ethanol, methanol, DMF and DMSO after a 3-h incubation at 25 °C. Less than 20% of activity was found in acetone and acetonitrile. The results show that rLac1 is thermostable, pH-stable and tolerant to organic solvents. These properties suggest that rLac1 has potential uses in industrial applications.

Keywords
ABTS, 2,2-azinobis-3-ethylbenzthiaoline-6-sulfonic acid; BSA, bovine serum albumin; EDTA, ethylenediaminetetraacetic acid; DMF, dimethylformamide; DMP, 2,6-dimethoxyphenol; DMSO, dimethylsulfoxide; DTT, dithiothreitol; TGA, thioglycolic acidpH stable; The
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Overexpression and characterization of a thermostable, pH-stable and organic solvent-tolerant Ganoderma fornicatum laccase in Pichia pastoris
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Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 46, Issue 7, July 2011, Pages 1469–1474
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