Summary
Biosynthesis of candicidin byStreptomyces acrimycini JI2236 was strongly inhibited by phosphate.p-Aminobenzoic acid (PABA) synthase activity, required for the synthesis of PABA, a candicindin precursor, was reduced by 72% in cells grown in medium supplemented with 7.5 mM phosphate. Hybridization studies showed that the DNA region ofS. acrimycini carrying thepabAB gene (encoding PABA synthase) is very similar to the homologous region ofS. griseus 3570.S. acrimycini was easily transformed with plasmids containing thepabAB gene ofS. griseus. Four transformants were studied in detail; three of the transformants synthesized higher levels of PABA synthase and two transformants produced more candicidin than control cultures transformed with pIJ699. The fourth transformant was unable to synthesize the antibiotic. Formation of PABA synthase and candicidin production was equally sensitive to phosphate regulation in transformants with thepabAB than in the untransformedS. acrimycini strain.
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References
Asturias, J.A., P. Liras and J.F. Martín. 1990. Phosphate control ofpabS gene transcription during candicidin biosynthesis. Gene 93: 79–84
Criado, L.M., J.F. Martín and J.A. Gil. 1993. Genepab ofStreptomyces griseus encoding a bifunctionalp-aminobenzoic acid synthase, is located between genes that may be involved in candicidin biosynthesis. Gene 126: 135–139.
Deng, Z., T. Kieser and D.A. Hopwood. 1988. ‘Strong incompatibility’ between derivatives of theStreptomyces multicopy plasmid pIJ101. Mol. Gen. Genet. 254: 286–294.
García-Domínguez, M., P. Liras and J.F. Martín. 1991. Cloning and characterization of the isopenicillin N synthase gene ofStreptomyces griseus NRRL 3851 and studies of expression and complementation of the cephamycin pathway inStreptomyces clavuligerus. Antimicrob. Agents Chemother. 35: 44–52.
Gil, J.A. and D.A. Hopwood. 1983. Cloning and expression of ap-aminobenzoic acid synthase gene of the candicidin producingStreptomyces griseus. Gene 25: 119–132.
Gil, J.A., L.M. Criado, M.T. Alegre and J.F. Martín. 1990. Use of a cloned gene involved in candicidin production to discover new polyene producerStreptomyces strains. FEMS Microbiol. Lett. 70: 15–18.
Gil, J.A., G. Naharro, J.R. Villanueva and J.F. Martín. 1985. Characterization and regulation ofp-aminobenzoic acid synthase fromStreptomyces griseus. J. Gen. Microbiol. 131: 1279–1287.
Hopwood, D.A., M.J. Bibb, K.F. Chater, T. Kieser, C.J. Bruton, H.M. Kieser, D.J. Lydiate, C.P. Smith, J.M. Ward and H. Schrempf. 1985. Genetic Manipulation ofStreptomyces. A Laboratory Manual. The John Innes Foundation. Norwich, UK.
Kięser, T. and R.E. Melton. 1988. Plasmid pIJ699, a multicopy positive-selection vector forStreptomyces. Gene 65: 83–91.
Langridge, J., P. Langridge and P.L. Bergquist. 1980. Extraction of nucleic acids from agarose gels. Anal. Biochem. 103: 264–271.
Lechevalier, H., R.F. Acker, C.T. Corke, C.M. Haenseler and S.A. Waksman. 1953. Candicidin, a new antifungal antibiotic. Mycologia 45: 155–171.
Liras, P., J.R. Villanueva and J.F. Martín. 1977. Sequential expression of macromolecule biosynthesis and candicidin formation inStreptomyces griseus. J. Gen. Microbiol. 102: 269–277.
Martín, J.F. and A.L. Demain. 1976. Control by phosphate of candicidin biosynthesis. Biochem. Biophys. Res. Commun. 71: 1103–1109.
Martín, J.F. and L.E. McDaniel. 1975. Kinetics of biosynthesis of polyene macrolide antibiotics in batch cultures: cell maturation time. Biotechnol. Bioeng. 17: 925–938.
Rebollo, A., J.A. Gil, P. Liras, J.A. Asturias and J.F. Martín. 1989. Cloning and characterization of a phosphate-regulated promoter involved in phosphate control of candicidin biosynthesis. Gene 79: 47–58.
Sambrook, J., E.F. Fritsch and T. Maniatis. 1989. Molecular Cloning. A Laboratory Manual, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Shaw, W.V. and D.A. Hopwood. 1976. Chloramphenicol acetylation inStreptomyces. J. Gen. Microbiol. 94: 159–166.
Zielinski, J., H. Borowy-Borowski, J. Golik, J. Gurrieriak, T. Zimisky, P. Kolodzi-Ejczyk, J. Pawlak and J. Borowski. 1979. The structure of levorin A2 and candicidin D. Tetrahedron. Lett. 20: 1791–1794.
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Asturias, J.A., Martín, J.F. & Liras, P. Biosynthesis and phosphate control of candicidin byStreptomyces acrimycini JI2236: effect of amplification of thepabAB gene. Journal of Industrial Microbiology 13, 183–189 (1994). https://doi.org/10.1007/BF01584005
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DOI: https://doi.org/10.1007/BF01584005