Cloning and characterization of a fish microsomal epoxide hydrolase of Danio rerio and application to kinetic resolution of racemic styrene oxide
| Autor: | Jae Woong Hwang, Sunghoon Park, Hee Sook Kim, Soojung Lee, Sang Jin Kim, Eun Yeol Lee, Eun Jung Lee |
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| Rok vydání: | 2005 |
| Předmět: |
animal structures
biology Stereochemistry Process Chemistry and Technology Danio Bioengineering biology.organism_classification Biochemistry Catalysis Kinetic resolution chemistry.chemical_compound Enantiopure drug chemistry Microsomal epoxide hydrolase Styrene oxide Oxyanion hole Epoxide hydrolase Enantiomeric excess |
| Zdroj: | Journal of Molecular Catalysis B: Enzymatic. 37:30-35 |
| ISSN: | 1381-1177 |
| Popis: | Gene mining of the genome database of the zebra fish Danio rerio revealed the presence of a putative microsomal epoxide hydrolase (mEH)-like protein containing the characteristic catalytic triad composed of Asp 223 , Glu 402 , and His 429 as well as the oxyanion hole common to all mEH. Based on the sequence information, a new EH gene was cloned by PCR amplification of cDNA of the zebra fish Danio rerio and expressed heterologously in Escherichia coli . The recombinant E. coli exhibited the enantiopreference toward ( R )-styrene oxide with the maximum hydrolytic activity of 11.4 μmol min −1 (mg dcw) −1 . When the kinetic resolution was conducted with 40 mM of racemic styrene oxide, enantiopure ( S )-styrene oxide was obtained with an enantiomeric excess (ee) higher than 99 and 23.5% yield at 30 min. These results demonstrate that the recombinant fish EH has the possible application as a biocatalyst for the production of enantiopure epoxides. |
| Databáze: | OpenAIRE |
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