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Complete Reconstitution of a Highly Reducing Iterative Polyketide Synthase
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Complete Reconstitution of a Highly Reducing Iterative Polyketide Synthase
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Journal Article
Complete Reconstitution of a Highly Reducing Iterative Polyketide Synthase
2009
Overview
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.
Publisher
American Association for the Advancement of Science,The American Association for the Advancement of Science
Subject
/ Biological and medical sciences
/ Enzymes
/ Fundamental and applied biological sciences. Psychology
/ Fungal Proteins - metabolism
/ Fungi
/ Lactones
/ Malonyl Coenzyme A - metabolism
/ Methods. Procedures. Technologies
/ Multienzyme Complexes - metabolism
/ Polyketide Synthases - chemistry
/ Polyketide Synthases - genetics
/ Polyketide Synthases - isolation & purification
/ Polyketide Synthases - metabolism
/ Pyrones
/ Recombinant Proteins - chemistry
/ Recombinant Proteins - isolation & purification
/ Recombinant Proteins - metabolism
/ S-Adenosylmethionine - metabolism
/ Saccharomyces cerevisiae - enzymology
/ Saccharomyces cerevisiae - genetics
/ Yeast
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