| Heterologous expression and biochemical characterization of acetyl xylan esterase from Coprinopsis cinerea. | |
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MedLine Citation:
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PMID: 23180549 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Acetyl xylan esterase (AXE) from basidiomycete Coprinopsis cinerea Okayama 7 (#130) was functionally expressed in Pichia pastoris with a C-terminal tag under the alcohol oxidase 1 (AOX1) promoter and secreted into the medium at 1.5 mg l(-1). Its molecular mass was estimated to be 65.5 kDa based on the SDS-PAGE analysis, which is higher than the calculated molecular mass of 40 kDa based on amino acid composition. In-silico analysis of the amino acid sequence predicted two potential N-glycosylation sites. Results from PNGase F deglycosylation and mass spectrum confirmed the presence of N-glycosylation on the recombinant AXE with predominant N-glycans HexNAc(2)Hex(9-16). The recombinant AXE showed best activity at 40 °C and pH 8. It showed not only acetyl esterase activity with a K(m) of 4.3 mM and a V(max) of 2.15 U mg(-1) for hydrolysis of 4-nitrophenyl acetate but also a butyl esterase activity for hydrolysis of 4-nitrophenyl butyrate with a K(m) of 0.11 mM and V(max) of 0.78 U mg(-1). The presence of two additional amino acid residues at its native N-terminus was found to help stabilize the enzyme against the protease cleavages without affecting its activity. |
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Authors:
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Veeresh Juturu; Christina Aust; Jin Chuan Wu |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2012-11-22 |
Journal Detail:
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Title: World journal of microbiology & biotechnology Volume: - ISSN: 1573-0972 ISO Abbreviation: World J. Microbiol. Biotechnol. Publication Date: 2012 Nov |
Date Detail:
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Created Date: 2012-11-27 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9012472 Medline TA: World J Microbiol Biotechnol Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Affiliation:
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Institute of Chemical and Engineering Sciences, Agency for Science, Technology and Research (A*STAR), 1 Pesek Road, Jurong Island, 627833, Singapore. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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