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Trehalose-6-phosphate synthase 1 from Metarhizium anisopliae: clone, expression and properties of the recombinant

Paper ID Volume ID Publish Year Pages File Format Full-Text
22177 43259 2009 7 PDF Available
Title
Trehalose-6-phosphate synthase 1 from Metarhizium anisopliae: clone, expression and properties of the recombinant
Abstract

Trehalose, an important component in fungal spores, is a disaccharide which protects against several environmental stresses, such as heat, desiccation, salt. Trehalose-6-phosphate synthase 1 (TPS1) is a subunit of trehalose synthase complex in fungi; it plays a key role in the biosynthesis of trehalose. In this study, a full-length cDNA from Metarhizium anisopliae encoding TPS1 (designated as MaTPS1) was isolated. The MaTPS1 cDNA is composed of 1836 nucleotides encoding a protein of 517 amino acids with a molecular mass of 58 kDa. The amino acid sequence has a relatively high homology with the TPS1s in several other filamentous fungi. Southern blot analysis showed that MaTPS1 gene occurs as a single copy in the M. anisopliae genome. And MaTPS1 was cloned into Pichia pastoris KM71 and secretively expressed with a histamine tag to facilitate a rapid purification of recombinant MaTPS1 (designated reTPS1). The properties of reTPS1 were examined. The Km value of reTPS1 for UDP-glucose and glucose-6-phosphate was 9.6 mM and 3.9 mM, respectively, and the optimal pH and temperature were about 6.5 and 40 °C. The enzyme was highly specific to glucose-6-phosphate for glucosyl acceptor, and its activity decreased rapidly as the concentrations of phosphate increased. The expression system will provide sufficient amounts of reTPS1 for future structural characterization of the protein and use for further investigation of MaTPS1's function.

Keywords
Metarhizium anisopliae; Trehalose-6-phosphate synthase 1; Trehalose; Pichia pastoris; Recombinant protein
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Trehalose-6-phosphate synthase 1 from Metarhizium anisopliae: clone, expression and properties of the recombinant
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 107, Issue 5, May 2009, Pages 499–505
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