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Molecular basis of Celmer's rules: stereochemistry of catalysis by isolated ketoreductase domains from modular polyketide synthases.

Siskos, Alexandros P. and Baerga-Ortiz, Abel and Bali, Shilpa and Stein, Viktor and Mamdani, Hassan and Spiteller, Dieter and Popovic, Bojana and Spencer, Jonathan B. and Staunton, James and Weissman, Kira J. and Leadlay, Peter F. (2005):
Molecular basis of Celmer's rules: stereochemistry of catalysis by isolated ketoreductase domains from modular polyketide synthases.
In: Chemistry & biology, pp. 1145-53, 12, (10), ISSN 1074-5521,
[Article]

Abstract

A system is reported for the recombinant expression of individual ketoreductase (KR) domains from modular polyketide synthases (PKSs) and scrutiny of their intrinsic specificity and stereospecificity toward surrogate diketide substrates. The eryKR(1) and the tylKR(1) domains, derived from the first extension module of the erythromycin PKS and the tylosin PKS, respectively, both catalyzed reduction of (2R, S)-2-methyl-3-oxopentanoic acid N-acetylcysteamine thioester, with complete stereoselectivity and stereospecificity, even though the substrate is not tethered to an acyl carrier protein or an intact PKS multienzyme. In contrast, and to varying degrees, the isolated enzymes eryKR(2), eryKR(5), and eryKR(6) exercised poorer control over substrate selection and the stereochemical course of ketoreduction. These data, together with modeling of diketide binding to KR(1) and KR(2), demonstrate the fine energetic balance between alternative modes of presentation of ketoacylthioester substrates to KR active sites.

Item Type: Article
Erschienen: 2005
Creators: Siskos, Alexandros P. and Baerga-Ortiz, Abel and Bali, Shilpa and Stein, Viktor and Mamdani, Hassan and Spiteller, Dieter and Popovic, Bojana and Spencer, Jonathan B. and Staunton, James and Weissman, Kira J. and Leadlay, Peter F.
Title: Molecular basis of Celmer's rules: stereochemistry of catalysis by isolated ketoreductase domains from modular polyketide synthases.
Language: English
Abstract:

A system is reported for the recombinant expression of individual ketoreductase (KR) domains from modular polyketide synthases (PKSs) and scrutiny of their intrinsic specificity and stereospecificity toward surrogate diketide substrates. The eryKR(1) and the tylKR(1) domains, derived from the first extension module of the erythromycin PKS and the tylosin PKS, respectively, both catalyzed reduction of (2R, S)-2-methyl-3-oxopentanoic acid N-acetylcysteamine thioester, with complete stereoselectivity and stereospecificity, even though the substrate is not tethered to an acyl carrier protein or an intact PKS multienzyme. In contrast, and to varying degrees, the isolated enzymes eryKR(2), eryKR(5), and eryKR(6) exercised poorer control over substrate selection and the stereochemical course of ketoreduction. These data, together with modeling of diketide binding to KR(1) and KR(2), demonstrate the fine energetic balance between alternative modes of presentation of ketoacylthioester substrates to KR active sites.

Journal or Publication Title: Chemistry & biology
Volume: 12
Number: 10
Divisions: 10 Department of Biology
10 Department of Biology > Protein Engineering of Ion Conducting Nanopores
Date Deposited: 14 Nov 2016 11:58
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