Abstract
Streptomyces castaneoglobisporus HUT6202 overproduces a diffusible melanin-like pigment. An operon, designated mel, containing a gene that encodes tyrosinase, which is involved in the synthesis of melanin pigment, was cloned from the chromosomal DNA of the microorganism into the high-copy plasmid pAK114 and expressed in S. lividans. The tyrosinase activity of the transformed cells was at approximately a 110-fold higher level than that of the same host carrying the plasmid pIJ702, which has the same replication origin as pAK114 and carries the mel operon from S. antibioticus. The sequence analysis of the S. castaneoglobisporus mel operon revealed that an open-reading frame consisting of 378 base pairs(bp), designated ORF378, was found upstream of the tyrosinase gene (TYRC) consisting of 819 bp. In the present study, we constructed a chimeric mel operon consisting of ORF378 from S. castaneoglobisporus and the tyrosinase gene (TYRA) from S. antibioticus. The chimeric mel operon or the S. antibioticus mel operon, which consists of ORF438 and TYRA, expressed the tyrosinase activity in Escherichia coli intracellularly when located under the control of lacZ promoter, and the tyrosinase activity from the former was at a 30-fold higher level than that from the latter. This suggests that the gene contributing to the high expression of the tyrosinase activity in S. castaneoglobisporus is ORF378, rather than TYRC.
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Bernan V, Filpula D, Herber W, Bibb M, Katz E (1985) The nucleotide sequence of the tyrosinase gene from Streptomyces antibioticus and characterization of the gene product. Gene 37: 101–110
Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248–254
Chen L-Y, Leu W-M, Wang K-T. Lee Y-HW (1992) Copper transfer and activation of the Streptomyces apotyrosinase are mediated through a complex formation between apotyrosinase and its trans-activator MelC1. J Biol Chem 267: 20100–20107
Ettlinger L, Corbaz R, Hütter R (1958) Zur Systematik der actinomycetcn. 4. Eine artereinteilung der Gattung Streptomyces Waksmann et Henrici. Arch Microbiol 31: 326–358
Hintermann G, Zatchej M, Hütter R (1985) Cloning and expression of the genetically unstable tyrosinase structural gene from Streptomyces glaucescens. Mol Gen Genet 200: 422–432
Hopwood DA, Bibb MJ, Chater KF, Kieser T, Bruton CJ, Kieser HM, Lydiate DJ, Smith CP, Ward JM, Schrempf H (1985) Genetic manipulation of Streptomyces: a laboratory manual. The John Innes Foundation, Norwich, UK
Huber M, Hintermann G, Lerch K (1985) Primary structure of tyrosinase from Streptomyces glaucescens. Biochemistry 24: 6038–6044
Katz E, Betancourt A (1988) Induction of tyrosinase by L-methionine in Streptomyces antibioticus. Can J Microbiol 34: 1297–1303
Katz E, Thompson CJ, Hopwood DA (1983) Cloning and expression of the tyrosinase gene from Streptomyces antibioticus in Streptomyces lividans. J Gen Microbiol 129: 2703–2714
Kawamoto S, Nakamura M, Yashima S (1993) Cloning, sequence and expression of the tyrosinase gene from Streptomyces lavendulae MA406 A-1. J Ferment Bioeng 76: 345–355
Küster E (1976) Chromogenecity of actinomycetes. In: Arai T(ed) Actinomycetes: the boundary microorganism Toppan, Tokyo. Japan, pp 43–54
Lee Y-H W. Chen B-F, Wu S-Y, Leu W-M, Lin J-J, Chen CW, Lo SJ (1988) A trans-acting gene is required for the phenotypic expression of a tyrosinase gene in Streptomyces. Gene 65: 71–81
Lerch K (1981) In: Sigel H (ed) Metal ions in biological systems vol 13. Dekker, New York, pp 143–184
Maniatis T, Fritsch EF, Sambrook J (1989) Molecular cloning. A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Murakami T, Holt TG, Thompson CJ (1989) Thiostrepton-induced gene expression in Streptomyces lividans. J Bacteriol 171: 1459–1466
Nishimura M, Sugiyama M (1994) Cloning and sequence analysis of a Streptomyces cholesterol esterase gene. Appl Microbiol Biotechnol 41: 419–424
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chainterminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467
Sugiyama M, Nomura H, Nimi O (1990) Use of the tyrosinase gene from Streptomyces to probe promoter sequences for Escherichia coli. Plasmid 23: 237–241
Thompson CJ, Ward JM, Hopwood DA (1982) Cloning of antibiotic resistance and nutritional genes in streptomycetes. J Bacteriol 151: 668–677
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Ikeda, K., Masujima, T., Suzuki, K. et al. Cloning and sequence analysis of the highly expressed melanin-synthesizing gene operon from Streptomyces castaneoglobisporus . Appl Microbiol Biotechnol 45, 80–85 (1996). https://doi.org/10.1007/s002530050652
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DOI: https://doi.org/10.1007/s002530050652