1t4m

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{{STRUCTURE_1t4m| PDB=1t4m | SCENE= }}
{{STRUCTURE_1t4m| PDB=1t4m | SCENE= }}
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'''STRUCTURE OF A THERMOSTABLE DOUBLE MUTANT OF BACILLUS SUBTILIS LIPASE OBTAINED THROUGH DIRECTED EVOLUTION'''
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===STRUCTURE OF A THERMOSTABLE DOUBLE MUTANT OF BACILLUS SUBTILIS LIPASE OBTAINED THROUGH DIRECTED EVOLUTION===
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==Overview==
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Variation in gene sequences generated by directed evolution approaches often does not assure a minimalist design for obtaining a desired property in proteins. While screening for enhanced thermostability, structural information was utilized in selecting mutations that are generated by error-prone PCR. By this approach we have increased the half-life of denaturation by 300-fold compared to the wild-type Bacillus subtilis lipase through three point mutations generated by only two cycles of error-prone PCR. At lower temperatures the activity parameters of the thermostable mutants are unaltered. High-resolution crystal structures of the mutants show subtle changes, which include stacking of tyrosine residues, peptide plane flipping and a better anchoring of the terminus, that challenge rational design and explain the structural basis for enhanced thermostability. The approach may offer an efficient and minimalist solution for the enhancement of a desired property of a protein.
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{{ABSTRACT_PUBMED_15321721}}
==About this Structure==
==About this Structure==
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[[Category: Sankaranarayanan, R.]]
[[Category: Sankaranarayanan, R.]]
[[Category: Alpha/beta hydrolase]]
[[Category: Alpha/beta hydrolase]]
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Sat May 3 09:31:12 2008''
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''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Mon Jul 28 02:28:18 2008''

Revision as of 23:28, 27 July 2008

Image:1t4m.png

Template:STRUCTURE 1t4m

STRUCTURE OF A THERMOSTABLE DOUBLE MUTANT OF BACILLUS SUBTILIS LIPASE OBTAINED THROUGH DIRECTED EVOLUTION

Template:ABSTRACT PUBMED 15321721

About this Structure

1T4M is a Single protein structure of sequence from Bacillus subtilis. Full crystallographic information is available from OCA.

Reference

Structural basis of selection and thermostability of laboratory evolved Bacillus subtilis lipase., Acharya P, Rajakumara E, Sankaranarayanan R, Rao NM, J Mol Biol. 2004 Aug 27;341(5):1271-81. PMID:15321721

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