Document Detail


Engineering the substrate specificity of the DhbE adenylation domain by yeast cell surface display.
MedLine Citation:
PMID:  23352143     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The adenylation (A) domains of nonribosomal peptide synthetases (NRPSs) activate aryl acids or amino acids to launch their transfer through the NRPS assembly line for the biosynthesis of many medicinally important natural products. In order to expand the substrate pool of NRPSs, we developed a method based on yeast cell surface display to engineer the substrate specificities of the A-domains. We acquired A-domain mutants of DhbE that have 11- and 6-fold increases in k(cat)/K(m) with nonnative substrates 3-hydroxybenzoic acid and 2-aminobenzoic acid, respectively and corresponding 3- and 33-fold decreases in k(cat)/K(m) values with the native substrate 2,3-dihydroxybenzoic acid, resulting in a dramatic switch in substrate specificity of up to 200-fold. Our study demonstrates that yeast display can be used as a high throughput selection platform to reprogram the "nonribosomal code" of A-domains.
Authors:
Keya Zhang; Kathryn M Nelson; Karan Bhuripanyo; Kimberly D Grimes; Bo Zhao; Courtney C Aldrich; Jun Yin
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Chemistry & biology     Volume:  20     ISSN:  1879-1301     ISO Abbreviation:  Chem. Biol.     Publication Date:  2013 Jan 
Date Detail:
Created Date:  2013-01-28     Completed Date:  2013-07-09     Revised Date:  2014-01-27    
Medline Journal Info:
Nlm Unique ID:  9500160     Medline TA:  Chem Biol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  92-101     Citation Subset:  IM    
Copyright Information:
Copyright © 2013 Elsevier Ltd. All rights reserved.
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MeSH Terms
Descriptor/Qualifier:
Bacillus subtilis / chemistry,  genetics*,  metabolism*
Catalytic Domain
Cell Surface Display Techniques / methods*
Hydroxybenzoates / metabolism
Mutation
Peptide Synthases / chemistry,  genetics*,  metabolism*
Substrate Specificity
Yeasts / cytology,  genetics*
ortho-Aminobenzoates / metabolism
Grant Support
ID/Acronym/Agency:
AI070219/AI/NIAID NIH HHS; R01 AI070219/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Hydroxybenzoates; 0/ortho-Aminobenzoates; 118-92-3/anthranilic acid; 303-38-8/2,3-dihydroxybenzoic acid; 99-06-9/3-hydroxybenzoic acid; EC 6.3.2.-/Peptide Synthases
Comments/Corrections
Comment In:
Chem Biol. 2013 Jan 24;20(1):3-5   [PMID:  23352133 ]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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