Employing Modular Polyketide Synthase Ketoreductases as Biocatalysts in the Preparative Chemoenzymatic Syntheses of Diketide Chiral Building Blocks

• Published in: Chemistry & biology Vol. 18; no. 10; pp. 1331 - 1340
• Main Authors: Piasecki, Shawn K., Taylor, Clint A., Detelich, Joshua F., Liu, June, Zheng, Jianting, Komsoukaniants, Arkady, Siegel, Dionicio R., Keatinge-Clay, Adrian T.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Ltd 28.10.2011
• Subjects: Alcohol Oxidoreductases - chemistry; Alcohol Oxidoreductases - genetics; Alcohol Oxidoreductases - metabolism; Bacterial Proteins - chemistry; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Biotechnology - methods; Catalysis; Cysteamine - analogs & derivatives; Cysteamine - chemistry; NADP - metabolism; Polyketide Synthases - chemistry; Polyketide Synthases - metabolism; Polyketides - chemical synthesis; Stereoisomerism
• ISSN: 1074-5521; 1879-1301
• DOI: 10.1016/j.chembiol.2011.07.021

Chiral building blocks are valuable intermediates in the syntheses of natural products and pharmaceuticals. A scalable chemoenzymatic route to chiral diketides has been developed that includes the general synthesis of α-substituted, β-ketoacyl N-acetylcysteamine thioesters followed by a biocatalytic cycle in which a glucose-fueled NADPH-regeneration system drives reductions catalyzed by isolated modular polyketide synthase (PKS) ketoreductases (KRs). To identify KRs that operate as active, stereospecific biocatalysts, 11 isolated KRs were incubated with 5 diketides and their products were analyzed by chiral chromatography. KRs that naturally reduce small polyketide intermediates were the most active and stereospecific toward the panel of diketides. Several biocatalytic reactions were scaled up to yield more than 100 mg of product. These syntheses demonstrate the ability of PKS enzymes to economically and greenly generate diverse chiral building blocks on a preparative scale. [Display omitted] ► Chiral analysis of reactions by ketoreductases on a panel of substrates ► Biocatalytically robust enzymes were identified for each ketoreductase type ► Glucose-driven reactions in lysate yielding >100 mg product ► General chemoenzymatic synthesis of chiral diketides for synthesis and enzymology