1zoi
From Proteopedia
(New page: 200px<br /><applet load="1zoi" size="350" color="white" frame="true" align="right" spinBox="true" caption="1zoi, resolution 1.6Å" /> '''Crystal Structure of ...) |
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==Overview== | ==Overview== | ||
| - | Esterase (EST) from Pseudomonas putida IFO12996 catalyzes the | + | 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. |
==About this Structure== | ==About this Structure== | ||
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[[Category: Pseudomonas putida]] | [[Category: Pseudomonas putida]] | ||
[[Category: Single protein]] | [[Category: Single protein]] | ||
| - | [[Category: Chen, C | + | [[Category: Chen, C J.]] |
| - | [[Category: Chen, Y | + | [[Category: Chen, Y J.]] |
[[Category: Elmi, F.]] | [[Category: Elmi, F.]] | ||
| - | [[Category: Hsieh, Y | + | [[Category: Hsieh, Y C.]] |
| - | [[Category: Huang, J | + | [[Category: Huang, J Y.]] |
| - | [[Category: Lee, H | + | [[Category: Lee, H T.]] |
| - | [[Category: Shaw, S | + | [[Category: Shaw, S Y.]] |
| - | [[Category: Wang, Y | + | [[Category: Wang, Y L.]] |
[[Category: alpha/beta hydrolase fold]] | [[Category: alpha/beta hydrolase fold]] | ||
[[Category: esterase]] | [[Category: esterase]] | ||
[[Category: pseudomonas putida]] | [[Category: pseudomonas putida]] | ||
| - | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on | + | ''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 16:17:36 2008'' |
Revision as of 14:17, 21 February 2008
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Crystal Structure of a Stereoselective Esterase from Pseudomonas putida IFO12996
Overview
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.
About this Structure
1ZOI is a Single protein structure of sequence from Pseudomonas putida. Full crystallographic information is available from OCA.
Reference
Stereoselective esterase from Pseudomonas putida IFO12996 reveals alpha/beta hydrolase folds for D-beta-acetylthioisobutyric acid synthesis., Elmi F, Lee HT, Huang JY, Hsieh YC, Wang YL, Chen YJ, Shaw SY, Chen CJ, J Bacteriol. 2005 Dec;187(24):8470-6. PMID:16321951
Page seeded by OCA on Thu Feb 21 16:17:36 2008
