Enantioselective ester hydrolase from Sphingobacterium sp. 238C5 useful for chiral resolution of β-phenylalanine and for its β-peptide synthesis
| Autor: | Jun Ogawa, Junichi Mano, Sakayu Shimizu, Tairo Hagishita |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
chemistry.chemical_classification
Sphingobacterium biology Chemistry Stereochemistry Process Chemistry and Technology Bioengineering Phenylalanine biology.organism_classification Biochemistry Esterase Catalysis Chiral resolution chemistry.chemical_compound Hydrolysis Stereospecificity Enzyme Peptide synthesis |
| Zdroj: | Journal of Molecular Catalysis B: Enzymatic. 60:138-144 |
| ISSN: | 1381-1177 |
| DOI: | 10.1016/j.molcatb.2009.04.011 |
| Popis: | A novel enzyme, β-phenylalanine ester hydrolase, useful for chiral resolution of β-phenylalanine and for its β-peptide synthesis was characterized. The enzyme purified from the cell free-extract of Sphingobacterium sp. 238C5 well hydrolyzed β-phenylalanine esters (S)-stereospecifically. Besides β-phenylalanine esters, the enzyme catalyzed the hydrolysis of several α-amino acid esters with l -stereospecificity, while the deduced 369 amino acid sequence of the enzyme exhibited homology to alkaline d -stereospecific peptide hydrolases from Bacillus strains. Escherichia coli transformant expressing the β-phenylalanine ester hydrolase gene exhibited an about 8-fold increase in specific (S)-β-phenylalanine ethyl ester hydrolysis as compared with that of Sphingobacterium sp. 238C5. The E. coli transformant showed (S)-enantiomer specific esterase activity in the reaction with a low concentration (30 mM) of β-phenylalanine ethyl ester, while it showed both esterase and transpeptidase activity in the reaction with a high concentration (170 mM) of β-phenylalanine ethyl ester and produced β-phenylalanyl-β-phenylalanine ethyl ester. This transpeptidase activity was useful for β-phenylalanine β-peptide synthesis. |
| Databáze: | OpenAIRE |
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