title: Identification of polyketide inhibitors targeting 3-dehydroquinate dehydratase in the shikimate pathway of Enterococcus faecalis.
abstract: Due to the emergence of resistance toward current antibiotics, there is a pressing need to develop the next generation of antibiotics as therapeutics against infectious and opportunistic diseases of microbial origins. The shikimate pathway is exclusive to microbes, plants and fungi, and hence is an attractive and logical target for development of antimicrobial therapeutics. The Gram-positive commensal microbe, Enterococcus faecalis, is a major human pathogen associated with nosocomial infections and resistance to vancomycin, the "drug of last resort". Here, we report the identification of several polyketide-based inhibitors against the E. faecalis shikimate pathway enzyme, 3-dehydroquinate dehydratase (DHQase). In particular, marein, a flavonoid polyketide, both inhibited DHQase and retarded the growth of Enterococcus faecalis. The purification, crystallization and structural resolution of recombinant DHQase from E. faecalis (at 2.2 Å resolution) are also reported. This study provides a route in the development of polyketide-based antimicrobial inhibitors targeting the shikimate pathway of the human pathogen E. faecalis.
authors: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
authors: Vivian Wing Ngar
authors: VW
authors: Cheung
authors: Institute of Molecular and Cell Biology, Singapore, Singapore.
authors: Bo
authors: B
authors: Xue
authors: Institute of Molecular and Cell Biology, Singapore, Singapore.
authors: Maria
authors: M
authors: Hernandez-Valladares
authors: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore.
authors: Maybelle Kho
authors: MK
authors: Go
authors: Institute of Molecular and Cell Biology, Singapore, Singapore.
authors: Alvin
authors: A
authors: Tung
authors: Institute of Molecular and Cell Biology, Singapore, Singapore.
authors: Adeleke H
authors: AH
authors: Aguda
authors: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore; Institute of Molecular and Cell Biology, Singapore, Singapore.
authors: Robert C
authors: RC
authors: Robinson
authors: Department of Biochemistry, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore; Synthetic Biology Research Consortium, National University of Singapore, Singapore, Singapore.
authors: Wen Shan
authors: WS
authors: Yew
mesh_terms: D000429:Alcohol Oxidoreductases / Q000235:genetics / Q000378:metabolism; D001665:Binding Sites; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D013293:Enterococcus faecalis / Q000201:enzymology* / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism; D005419:Flavonoids / Q000737:chemistry / Q000378:metabolism; D006836:Hydro-Lyases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D007700:Kinetics; D061065:Polyketides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D012765:Shikimic Acid / Q000737:chemistry / Q000378:metabolism*
chemical_list: D004791:Enzyme Inhibitors; D005419:Flavonoids; D061065:Polyketides; D011994:Recombinant Proteins; D012765:Shikimic Acid; D000429:Alcohol Oxidoreductases; C022641:Shikimate dehydrogenase; D006836:Hydro-Lyases; C020000:3-dehydroquinate dehydratase