Insights into the programmed ketoreduction of partially reducing polyketide synthases : stereo- and substrate-specificity of the ketoreductase domain

One of the hallmarks of iterative polyketide synthases (PKSs) is the programming mechanism which is essential for the generation of structurally diverse polyketide products. In partially reducing iterative PKSs (PR-PKSs), the programming mechanism is mainly dictated by the ketoreductase (KR) domain....

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Main Authors: Soehano, Ishin, Yang, Lifeng, Ding, Feiqing, Sun, Huihua, Low, Zhen Jie, Liu, Xuewei, Liang, Zhao-Xun
Other Authors: School of Biological Sciences
Format: Article
Language:English
Published: 2015
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Online Access:https://hdl.handle.net/10356/106999
http://hdl.handle.net/10220/25227
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-1069992023-02-28T17:04:22Z Insights into the programmed ketoreduction of partially reducing polyketide synthases : stereo- and substrate-specificity of the ketoreductase domain Soehano, Ishin Yang, Lifeng Ding, Feiqing Sun, Huihua Low, Zhen Jie Liu, Xuewei Liang, Zhao-Xun School of Biological Sciences School of Physical and Mathematical Sciences DRNTU::Science::Biological sciences::Molecular biology One of the hallmarks of iterative polyketide synthases (PKSs) is the programming mechanism which is essential for the generation of structurally diverse polyketide products. In partially reducing iterative PKSs (PR-PKSs), the programming mechanism is mainly dictated by the ketoreductase (KR) domain. The KR domain contributes to the programming of PR-PKSs through selective reduction of polyketide intermediates. How the KR domain achieves the selective ketoreduction remains to be fully understood. In this study, we found that the KR domain of the (R)-mellein-synthesizing PR-PKS SACE5532 functions as a B-type KR domain to generate (R)-hydroxyl functionalities. Comparative studies of the KR domains of SACE5532 and NcsB suggested that the two KR domains have distinct substrate preferences towards simple N-acetylcysteamine thioester (SNAC) substrates. We further found that the substrate preference of KRSACE5532 can be switched by swapping several motifs with KRNcsB, and that swapping of the same motifs in the full length SACE5532 resulted in a reprogramming of the PKS. Together, the results advance our understanding of the programming of iterative PR-PKSs by providing new support to the hypothesis that the programmed ketoreduction is accomplished by differential recognition of polyketide intermediates. Accepted version 2015-03-11T03:54:14Z 2019-12-06T22:22:48Z 2015-03-11T03:54:14Z 2019-12-06T22:22:48Z 2012 2012 Journal Article Soehano, I., Yang, L., Ding, F., Sun, H., Low, Z. J., Liu, X., et al. (2014). Insights into the programmed ketoreduction of partially reducing polyketide synthases : stereo- and substrate-specificity of the ketoreductase domain. Organic & biomolecular chemistry, 12(42), 8542-8549. https://hdl.handle.net/10356/106999 http://hdl.handle.net/10220/25227 10.1039/C4OB01777C en Organic & biomolecular chemistry © 2012 The Royal Society of Chemistry. This is the author created version of a work that has been peer reviewed and accepted for publication by Organic & Biomolecular Chemistry, The Royal Society of Chemistry. It incorporates referee’s comments but changes resulting from the publishing process, such as copyediting, structural formatting, may not be reflected in this document. The published version is available at: [http://dx.doi.org/10.1039/C4OB01777C]. 8 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science::Biological sciences::Molecular biology
spellingShingle DRNTU::Science::Biological sciences::Molecular biology
Soehano, Ishin
Yang, Lifeng
Ding, Feiqing
Sun, Huihua
Low, Zhen Jie
Liu, Xuewei
Liang, Zhao-Xun
Insights into the programmed ketoreduction of partially reducing polyketide synthases : stereo- and substrate-specificity of the ketoreductase domain
description One of the hallmarks of iterative polyketide synthases (PKSs) is the programming mechanism which is essential for the generation of structurally diverse polyketide products. In partially reducing iterative PKSs (PR-PKSs), the programming mechanism is mainly dictated by the ketoreductase (KR) domain. The KR domain contributes to the programming of PR-PKSs through selective reduction of polyketide intermediates. How the KR domain achieves the selective ketoreduction remains to be fully understood. In this study, we found that the KR domain of the (R)-mellein-synthesizing PR-PKS SACE5532 functions as a B-type KR domain to generate (R)-hydroxyl functionalities. Comparative studies of the KR domains of SACE5532 and NcsB suggested that the two KR domains have distinct substrate preferences towards simple N-acetylcysteamine thioester (SNAC) substrates. We further found that the substrate preference of KRSACE5532 can be switched by swapping several motifs with KRNcsB, and that swapping of the same motifs in the full length SACE5532 resulted in a reprogramming of the PKS. Together, the results advance our understanding of the programming of iterative PR-PKSs by providing new support to the hypothesis that the programmed ketoreduction is accomplished by differential recognition of polyketide intermediates.
author2 School of Biological Sciences
author_facet School of Biological Sciences
Soehano, Ishin
Yang, Lifeng
Ding, Feiqing
Sun, Huihua
Low, Zhen Jie
Liu, Xuewei
Liang, Zhao-Xun
format Article
author Soehano, Ishin
Yang, Lifeng
Ding, Feiqing
Sun, Huihua
Low, Zhen Jie
Liu, Xuewei
Liang, Zhao-Xun
author_sort Soehano, Ishin
title Insights into the programmed ketoreduction of partially reducing polyketide synthases : stereo- and substrate-specificity of the ketoreductase domain
title_short Insights into the programmed ketoreduction of partially reducing polyketide synthases : stereo- and substrate-specificity of the ketoreductase domain
title_full Insights into the programmed ketoreduction of partially reducing polyketide synthases : stereo- and substrate-specificity of the ketoreductase domain
title_fullStr Insights into the programmed ketoreduction of partially reducing polyketide synthases : stereo- and substrate-specificity of the ketoreductase domain
title_full_unstemmed Insights into the programmed ketoreduction of partially reducing polyketide synthases : stereo- and substrate-specificity of the ketoreductase domain
title_sort insights into the programmed ketoreduction of partially reducing polyketide synthases : stereo- and substrate-specificity of the ketoreductase domain
publishDate 2015
url https://hdl.handle.net/10356/106999
http://hdl.handle.net/10220/25227
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