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| {{STRUCTURE_1zoi| PDB=1zoi | SCENE= }} | | {{STRUCTURE_1zoi| PDB=1zoi | SCENE= }} |
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| '''Crystal Structure of a Stereoselective Esterase from Pseudomonas putida IFO12996'''
| | ===Crystal Structure of a Stereoselective Esterase from Pseudomonas putida IFO12996=== |
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| ==Overview==
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| Esterase (EST) from Pseudomonas putida IFO12996 catalyzes the stereoselective hydrolysis of methyl dl-beta-acetylthioisobutyrate (dl-MATI) to produce d-beta-acetylthioisobutyric acid (DAT), serving as a key intermediate for the synthesis of angiotensin-converting enzyme inhibitors. The EST gene was cloned and expressed in Escherichia coli; the recombinant protein is a non-disulfide-linked homotrimer with a monomer molecular weight of 33,000 in both solution and crystalline states, indicating that these ESTs function as trimers. EST hydrolyzed dl-MATI to produce DAT with a degree of conversion of 49.5% and an enantiomeric excess value of 97.2% at an optimum pH of about 8 to 10 and an optimum temperature of about 57 to 67 degrees C. The crystal structure of EST has been determined by X-ray diffraction to a resolution of 1.6 A, confirming that EST is a member of the alpha/beta hydrolase fold superfamily of enzymes and includes a catalytic triad of Ser97, Asp227, and His256. The active site is located approximately in the middle of the molecule at the end of a pocket approximately 12 A deep. EST can hydrolyze the methyl ester group without affecting the acetylthiol ester moiety in dl-MATI. The examination of substrate specificity of EST toward other linear esters revealed that the enzyme showed specific activity toward methyl esters and that it recognized the configuration at C-2.
| | The line below this paragraph, {{ABSTRACT_PUBMED_16321951}}, adds the Publication Abstract to the page |
| | (as it appears on PubMed at http://www.pubmed.gov), where 16321951 is the PubMed ID number. |
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| | {{ABSTRACT_PUBMED_16321951}} |
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| ==About this Structure== | | ==About this Structure== |
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| [[Category: Esterase]] | | [[Category: Esterase]] |
| [[Category: Pseudomonas putida]] | | [[Category: Pseudomonas putida]] |
| ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 17:52:45 2008'' | | |
| | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sun Jul 27 17:22:34 2008'' |
