| Enantioselective oxidation of 2-hydroxy carboxylic acids by glycolate oxidase and catalase coexpressed in methylotrophic Pichia pastoris. | |
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MedLine Citation:
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PMID: 20014430 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Glycolate oxidase (GO; (S)-2-hydroxyacid oxidase, EC 1.1.3.15) is a flavin mononucleotide (FMN)-dependent enzyme, which catalyzes the oxidation of 2-hydroxy carboxylic acids to the corresponding 2-keto acids. Catalase has been used as cocatalyst to decompose hydrogen peroxide produced in the reaction, thus limiting peroxide-based side reactions and GO deactivation. GO from spinach and catalase T from Saccharomyces cerevisiae previously coexpressed in Pichia pastoris strain NRRL Y-21001, was permeabilized and used for the oxidation of 3-phenyllactic acid, 3-indolelactic acid, 3-chlorolactic acid, 2-hydroxybutanoic acid, and 2-hydroxydecanoic acid to demonstrate high degree of selectivity to the (S)-enantiomers, leaving (R)-isomers intact. The rates of oxidation ranged from 1.3 to 120.0%, relative to the oxidation of lactic acid to pyruvic acid. The best substrates were 3-chlorolactic acid (110%) and 2-hydroxybutanoic acid (120%). Oxidation was carried out with (R)-, (S)-, and (RS)-3-phenyllactic acid, (RS)-lactic acid, and (RS)-2-hydroxybutanoic acid in 500 mL scale to characterize the products and stoichiometry of the reaction. All (RS)- and (S)-2-hydroxy acids produced 2-keto acids at close to the theoretical yield in 1-9 h. (R)-3-Phenyllactic acid was not oxidized over a period of 9 h. Addition of exogenous FMN and catalase were not required for this oxidation using double recombinant Pichia pastoris whole cells. As GO is absolutely specific to (S)-enantiomers, it can be used for resolution of racemic 2-hydroxy acids to (R)-2-hydroxy acids as well as for production of 2-keto acids. This is the first report on the selectivity of a broad range of 2-hydroxy acids by GO. |
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Authors:
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Shuvendu Das; James H Glenn; Mani Subramanian |
Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Biotechnology progress Volume: 26 ISSN: 1520-6033 ISO Abbreviation: Biotechnol. Prog. Publication Date: 2010 May-Jun |
Date Detail:
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Created Date: 2010-06-21 Completed Date: 2010-10-04 Revised Date: 2012-05-23 |
Medline Journal Info:
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Nlm Unique ID: 8506292 Medline TA: Biotechnol Prog Country: United States |
Other Details:
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Languages: eng Pagination: 607-15 Citation Subset: IM |
Copyright Information:
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Copyright 2009 American Institute of Chemical Engineers |
Affiliation:
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Center for Biocatalysis and Bioprocessing, The University of Iowa, Iowa City, IA 52242, USA. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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2,6-Dichloroindophenol Alcohol Oxidoreductases / biosynthesis, genetics, metabolism* Benzalkonium Compounds Biomass Catalase / biosynthesis, genetics, metabolism* Cell Membrane Permeability Chromatography, High Pressure Liquid Fermentation Hydroxy Acids / metabolism* Hydroxybutyrates / metabolism Lactic Acid / metabolism Oxidation-Reduction Oxygen / metabolism Pichia / chemistry, enzymology*, genetics Pyruvic Acid / metabolism Stereoisomerism |
| Chemical | |
Reg. No./Substance:
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0/Benzalkonium Compounds; 0/Hydroxy Acids; 0/Hydroxybutyrates; 127-17-3/Pyruvic Acid; 50-21-5/Lactic Acid; 565-70-8/2-hydroxybutyric acid; 7782-44-7/Oxygen; 956-48-9/2,6-Dichloroindophenol; EC 1.1.-/Alcohol Oxidoreductases; EC 1.1.3.15/glycollate oxidase; EC 1.11.1.6/Catalase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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