Expression and characterization of the type III polyketide synthase 1,3,6,8-tetrahydroxynaphthalene synthase from Streptomyces coelicolor A3(2)
- 1 August 2003
- journal article
- research article
- Published by Oxford University Press (OUP) in Journal of Industrial Microbiology & Biotechnology
- Vol. 30 (8) , 510-515
- https://doi.org/10.1007/s10295-003-0075-8
Abstract
Sequence analysis of the metabolically rich 8.7-Mbp genome of the model actinomycete Streptomyces coelicolor A3(2) revealed three genes encoding predicted type III polyketide synthases (PKSs). We report the inactivation, expression, and characterization of the type III PKS homologous SCO1206 gene product as 1,3,6,8-tetrahydroxynaphthalene synthase (THNS). Incubation of recombinant THNS with malonyl-CoA showed THN production, as demonstrated by UV and HPLC analyses. The Km value for malonyl-CoA and the kcat value for THN synthesis were determined spectrophotometrically to be 3.58±0.85 µM and 0.48±0.03 min−1, respectively. The C-terminal region of S. coelicolor THNS, which is longer than most other bacterial and plant type III PKSs, was shortened by 25 amino acid residues and the resulting mutant was shown to be slightly more active (Km=1.97±0.19 µM, kcat=0.75±0.04 min−1) than the wild-type enzyme.Keywords
This publication has 21 references indexed in Scilit:
- Alteration of reaction and substrate specificity of a bacterial type III polyketide synthase by site-directed mutagenesisBiochemical Journal, 2002
- Identification and cloning of a type III polyketide synthase required for diffusible pigment biosynthesis in Saccharopolyspora erythraea‡Molecular Microbiology, 2002
- Complete genome sequence of the model actinomycete Streptomyces coelicolor A3(2)Nature, 2002
- Discovery of a New Bacterial Polyketide Biosynthetic PathwayChemBioChem, 2001
- Genetic Diversity of phlD from 2,4-Diacetylphloroglucinol-Producing Fluorescent Pseudomonas spp.Phytopathology®, 2001
- Functional analysis of genes from Streptomyces griseus involved in the synthesis of isorenieratene, a carotenoid with aromatic end groups, revealed a novel type of carotenoid desaturaseBiochimica et Biophysica Acta (BBA) - Molecular and Cell Biology of Lipids, 1999
- Heterologous Expression and Product Identification ofColletotrichum lagenariumPolyketide Synthase Encoded by thePKS1Gene Involved in Melanin BiosynthesisBioscience, Biotechnology, and Biochemistry, 1999
- Ectopic expression of the Streptomyces coelicolor whiE genes for polyketide spore pigment synthesis and their interaction with the act genes for actinorhodin biosynthesisMicrobiology, 1995
- Overexpression of a Gene Cluster Encoding a Chalcone Synthase-like Protein Confers Redbrown Pigment Production in Streptomyces griseus.The Journal of Antibiotics, 1995
- Gene disruption and gene replacement inStreptomycesvia single stranded DNA transformation of integration vectorsNucleic Acids Research, 1991