Enhanced FK506 production in Streptomyces clavuligerus CKD1119 by engineering the supply of methylmalonyl-CoA precursor
- 12 September 2009
- journal article
- Published by Oxford University Press (OUP) in Journal of Industrial Microbiology & Biotechnology
- Vol. 36 (12), 1473-1482
- https://doi.org/10.1007/s10295-009-0635-7
Abstract
FK506 is a 23-membered polyketide macrolide with immunosuppressant activity produced by Streptomyces species. The production of FK506 in S. clavuligerus CKD1119 (KCTC 10561BP) was improved by enhancing the supply of biosynthetic precursors. This improvement was approximately 2.5-fold (15 mg/l) with the supplementation of 10 mM methyl oleate, which is the probable source of acyl-CoAs, to R2YE medium. When the level of FK506 production reached its maximum, the intracellular concentration of methylmalonyl-CoA in S. clavuligerus CKD1119 supplemented with methyl oleate was 12.5-fold higher than that of the unsupplemented strain, suggesting that an increased methylmalonyl-CoA level caused the high-level production of FK506. The following three pathways for the production of (2S)-methylmalonyl-CoA were evaluated to identify the effective precursor supply pathway that can support the high production of FK506 in S. clavuligerus CKD1119: propionyl-CoA carboxylase, methylmalonyl-CoA mutase (MCM), and malonyl/methylmalonyl-CoA ligase. Of the three pathways examined, the MCM pathway supported the highest levels of FK506 production. The expression of MCM in S. clavuligerus CKD1119 led to a threefold and 1.5-fold increase in the methylmalonyl-CoA pool and FK506 production, respectively. Supplementing the culture broth of S. clavuligerus CKD1119 expressing MCM with methyl oleate resulted in an additional twofold increase in the FK506 titer (17.8 mg/l). Overall, these results show that the methylmalonyl-CoA supply is a limiting factor for FK506 biosynthesis and that among the three pathways analyzed, the MCM pathway is the most effective precursor supply pathway supporting the highest titer of FK506 in S. clavuligerus CKD1119.Keywords
This publication has 40 references indexed in Scilit:
- Isolation and identification of a novel microorganism producing the immunosuppressant tacrolimusJournal of Bioscience and Bioengineering, 2008
- Ascomycin and FK506: Pharmacology and Therapeutic Potential as Anticonvulsants and NeuroprotectantsCNS Neuroscience & Therapeutics, 2008
- Ascomycin and FK506: Pharmacology and Therapeutic Potential as Anticonvulsants and NeuroprotectantsCNS Neuroscience & Therapeutics, 2008
- Synthesis of C 5 -dicarboxylic acids from C 2 -units involving crotonyl-CoA carboxylase/reductase: The ethylmalonyl-CoA pathwayProceedings of the National Academy of Sciences, 2007
- Engineering of the methylmalonyl-CoA metabolite node of Saccharopolyspora erythraea for increased erythromycin productionMetabolic Engineering, 2007
- Propionyl-CoA carboxylase from Streptomyces coelicolor A3(2) : cloning of the gene encoding the biotinlcontaining subunitMicrobiology, 1996
- Effects of Exogenous Methyl Oleate on the Biosynthesis of Nigericin, a Polyether Carboxylic Antibiotic, by Streptomyces hygroscopicus NRRL B-1865Bioscience, Biotechnology, and Biochemistry, 1992
- Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexesCell, 1991
- Butyrate metabolism in streptomycetes. Characterization of an intramolecular vicinal interchange rearrangement linking isobutyrate and butyrate in Streptomyces cinnamonensisJournal of the Chemical Society, Perkin Transactions 1, 1988
- Methods for characterization of Streptomyces speciesInternational Journal of Systematic and Evolutionary Microbiology, 1966