Publication Date:
2009-11-11
Description:
Highly reducing iterative polyketide synthases are large, multifunctional enzymes that make important metabolites in fungi, such as lovastatin, a cholesterol-lowering drug from Aspergillus terreus. We report efficient expression of the lovastatin nonaketide synthase (LovB) from an engineered strain of Saccharomyces cerevisiae, as well as complete reconstitution of its catalytic function in the presence and absence of cofactors (the reduced form of nicotinamide adenine dinucleotide phosphate and S-adenosylmethionine) and its partner enzyme, the enoyl reductase LovC. Our results demonstrate that LovB retains correct intermediates until completion of synthesis of dihydromonacolin L, but off-loads incorrectly processed compounds as pyrones or hydrolytic products. Experiments replacing LovC with analogous MlcG from compactin biosynthesis demonstrate a gate-keeping function for this partner enzyme. This study represents a key step in the understanding of the functions and structures of this family of enzymes.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2875069/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉 〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2875069/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Ma, Suzanne M -- Li, Jesse W-H -- Choi, Jin W -- Zhou, Hui -- Lee, K K Michael -- Moorthie, Vijayalakshmi A -- Xie, Xinkai -- Kealey, James T -- Da Silva, Nancy A -- Vederas, John C -- Tang, Yi -- 1R01GM085128/GM/NIGMS NIH HHS/ -- 1R21GM077264/GM/NIGMS NIH HHS/ -- R01 GM085128/GM/NIGMS NIH HHS/ -- R01 GM085128-02/GM/NIGMS NIH HHS/ -- New York, N.Y. -- Science. 2009 Oct 23;326(5952):589-92. doi: 10.1126/science.1175602.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Department of Chemical and Biomolecular Engineering, University of California, Los Angeles, CA 90095, USA.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/19900898" target="_blank"〉PubMed〈/a〉
Keywords:
Aspergillus/enzymology/genetics/metabolism
;
Biocatalysis
;
Catalytic Domain
;
Cloning, Molecular
;
Fungal Proteins/metabolism
;
Ketones/metabolism
;
Lactones/metabolism
;
Lovastatin/biosynthesis
;
Malonyl Coenzyme A/metabolism
;
Molecular Structure
;
Multienzyme Complexes/metabolism
;
NAD/metabolism
;
Naphthalenes/*metabolism
;
Polyketide Synthases/chemistry/genetics/isolation & purification/*metabolism
;
Pyrones/metabolism
;
Recombinant Proteins/chemistry/isolation & purification/metabolism
;
S-Adenosylmethionine/metabolism
;
Saccharomyces cerevisiae/enzymology/*genetics
;
Substrate Specificity
Print ISSN:
0036-8075
Electronic ISSN:
1095-9203
Topics:
Biology
,
Chemistry and Pharmacology
,
Computer Science
,
Medicine
,
Natural Sciences in General
,
Physics
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