| A single Phe54Tyr substitution improves the catalytic activity and thermostability of Trigonopsis variabilis D-amino acid oxidase. | |
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MedLine Citation:
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PMID: 19909828 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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The industrial importance of Trigonopsis variabilisd-amino acid oxidase (TvDAAO) is represented by its biocatalytic oxidative deamination of cephalosporin C (CPC) to yield glutaryl-7-aminocephalosporanic acid (GL-7-ACA). The process has been incorporated into a two-step bioconversion to produce 7-aminocephalosporanic acid, the crucial synthetic nucleus for several semi-synthetic cephalosporin antibiotics. A homology model of TvDAAO indicated that residue F54 is in a close proximity to the in silico docked CPC. Substitution of this F54 to Tyr (F54Y) resulted in 6-fold improvement in k(cat,app) and approximately 2.5-fold increase in K(i) of GL-7-ACA. Heat treatment (55 degrees C, 60 min) did not decrease the activity of F54Y. It is suggested that the Tyr substitution might initiate hydrogen bond formation with the amino group of CPC and facilitate deamination. Faster substrate turnover, reduced GL-7-ACA inhibition and improved thermostability of the F54Y substitution render it a useful candidate in industrial production of semi-synthetic cephems. |
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Authors:
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Kin-Sing Wong; Wing-Ping Fong; Paul Wai-Kei Tsang |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2009-11-10 |
Journal Detail:
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Title: New biotechnology Volume: 27 ISSN: 1876-4347 ISO Abbreviation: N Biotechnol Publication Date: 2010 Feb |
Date Detail:
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Created Date: 2010-03-01 Completed Date: 2010-05-24 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 101465345 Medline TA: N Biotechnol Country: Netherlands |
Other Details:
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Languages: eng Pagination: 78-84 Citation Subset: IM |
Copyright Information:
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Copyright 2009 Elsevier B.V. All rights reserved. |
Affiliation:
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Department of Biochemistry, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Amino Acid Sequence Amino Acid Substitution* Animals Catalysis Cephalosporins / chemical synthesis, chemistry, metabolism D-Amino-Acid Oxidase* / chemistry, genetics, metabolism Enzyme Stability* Fungal Proteins* / genetics, metabolism Humans Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Phenylalanine / chemistry, genetics Protein Structure, Tertiary* Saccharomycetales / enzymology* Sequence Alignment Temperature Tyrosine / chemistry, genetics |
| Chemical | |
Reg. No./Substance:
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0/Cephalosporins; 0/Fungal Proteins; 27920-90-7/glutaryl-7-aminocephalosporanic acid; 55520-40-6/Tyrosine; 61-24-5/cephalosporin C; 63-91-2/Phenylalanine; 957-68-6/7-aminocephalosporanic acid; EC 1.4.3.3/D-Amino-Acid Oxidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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