1c7j

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[[Image:1c7j.gif|left|200px]]
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==Overview==
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{{ABSTRACT_10535917}}
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Two directed evolution experiments on p-nitrobenzyl esterase yielded one enzyme with a 100-fold increased activity in aqueous-organic solvents and another with a 17 degrees C increase in thermostability. Structures of the wild type and its organophilic and thermophilic counterparts are presented at resolutions of 1.5 A, 1.6 A, and 2.0 A, respectively. These structures identify groups of interacting mutations and demonstrate how directed evolution can traverse complex fitness landscapes. Early-generation mutations stabilize flexible loops not visible in the wild-type structure and set the stage for further beneficial mutations in later generations. The mutations exert their influence on the esterase structure over large distances, in a manner that would be difficult to predict. The loops with the largest structural changes generally are not the sites of mutations. Similarly, none of the seven amino acid substitutions in the organophile are in the active site, even though the enzyme experiences significant changes in the organization of this site. In addition to reduction of surface loop flexibility, thermostability in the evolved esterase results from altered core packing, helix stabilization, and the acquisition of surface salt bridges, in agreement with other comparative studies of mesophilic and thermophilic enzymes. Crystallographic analysis of the wild type and its evolved counterparts reveals networks of mutations that collectively reorganize the active site. Interestingly, the changes that led to diversity within the alpha/beta hydrolase enzyme family and the reorganization seen in this study result from main-chain movements.
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==About this Structure==
==About this Structure==
1C7J is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis]. This structure supersedes the now removed PDB entry [http://oca.weizmann.ac.il/oca-bin/send-pdb?obs=1&id=1qe8 1qe8]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1C7J OCA].
1C7J is a [[Single protein]] structure of sequence from [http://en.wikipedia.org/wiki/Bacillus_subtilis Bacillus subtilis]. This structure supersedes the now removed PDB entry [http://oca.weizmann.ac.il/oca-bin/send-pdb?obs=1&id=1qe8 1qe8]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1C7J OCA].
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==Reference==
 
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A structural view of evolutionary divergence., Spiller B, Gershenson A, Arnold FH, Stevens RC, Proc Natl Acad Sci U S A. 1999 Oct 26;96(22):12305-10. PMID:[http://www.ncbi.nlm.nih.gov/pubmed/10535917 10535917]
 
[[Category: Bacillus subtilis]]
[[Category: Bacillus subtilis]]
[[Category: Single protein]]
[[Category: Single protein]]
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[[Category: Organic activity]]
[[Category: Organic activity]]
[[Category: Pnb esterase]]
[[Category: Pnb esterase]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Fri May 2 12:25:21 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jun 25 19:21:09 2008''

Revision as of 16:21, 25 June 2008

Image:1c7j.png

Template:STRUCTURE 1c7j

PNB ESTERASE 56C8


Template:ABSTRACT 10535917

About this Structure

1C7J is a Single protein structure of sequence from Bacillus subtilis. This structure supersedes the now removed PDB entry 1qe8. Full crystallographic information is available from OCA.

Page seeded by OCA on Wed Jun 25 19:21:09 2008

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