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Purification, characterization and functional analysis of asparagines synthetase encoding by ste10 gene in Ebosin biosynthesis of Streptomyces sp. 139

Paper ID Volume ID Publish Year Pages File Format Full-Text
17778 42695 2008 6 PDF Available
Title
Purification, characterization and functional analysis of asparagines synthetase encoding by ste10 gene in Ebosin biosynthesis of Streptomyces sp. 139
Abstract

Ebosin produced by Streptomyces sp. 139 is a novel exopolysaccharide (EPS) with medicinal activity. This paper describes the functional study of ste10, a putative Ebosin biosynthesis gene. ste10 was cloned and expressed in Escherichia coli BL21 and the purified recombinant protein characterized. Ste10 was shown to be able of catalyzing the transfer of amide nitrogen of glutamine to the side chain of aspartate to produce asparagine. Its Km, optimum temperature and pH were determined to be 0.9 mM, 37 °C and 7.38, respectively. After ste10 gene knock-out, the monosaccharide composition of EPS-m produced by the mutant Streptomyces sp. 139 (ste10−) was found changed in comparison with that of Ebosin while its antagonist activity for IL-1R decreased significantly. Based on these results, it is concluded that ste10 codes for an asparagine synthetase which may function as a modificator gene of Ebosin during its biosynthesis.

Keywords
ELISA, enzyme-linked immunospecific assay; EPS, exopolysaccharide; GC, gas chromatography; HPLC, high-performance liquid chromatography; IL-1R, interleukin-1 receptor; IPTG, isopropyl-β-d-thiogalactopyranosideAsparagine synthetase; Ste10 gene expression;
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Purification, characterization and functional analysis of asparagines synthetase encoding by ste10 gene in Ebosin biosynthesis of Streptomyces sp. 139
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 42, Issue 7, 5 June 2008, Pages 548–553
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