Abstract
We have cloned and sequenced a 1.5-kb chromosomal fragment of Thermus thermophilus which promoted the overproduction of carotenoids in T. thermophilus. An open reading frame (ORF-A) coding for a polypeptide with 289 amino acids was responsible for carotenoid overproduction. The putative ORF-A protein showed significant homology with the amino acid sequences of crtB gene products (phytoene syntheses) of other microorganisms. The clone containing the ORF-A on a multicopy plasmid produced about three times as much carotenoid as that produced by the host strain, suggesting that the crtB gene product is a rate-limiting enzyme for carotenoid biosynthesis in T. thermophilus.
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Selected References
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- Armstrong G. A., Alberti M., Hearst J. E. Conserved enzymes mediate the early reactions of carotenoid biosynthesis in nonphotosynthetic and photosynthetic prokaryotes. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9975–9979. doi: 10.1073/pnas.87.24.9975. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Armstrong G. A., Alberti M., Leach F., Hearst J. E. Nucleotide sequence, organization, and nature of the protein products of the carotenoid biosynthesis gene cluster of Rhodobacter capsulatus. Mol Gen Genet. 1989 Apr;216(2-3):254–268. doi: 10.1007/BF00334364. [DOI] [PubMed] [Google Scholar]
- BLIGH E. G., DYER W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959 Aug;37(8):911–917. doi: 10.1139/o59-099. [DOI] [PubMed] [Google Scholar]
- Hishinuma F., Tanaka T., Sakaguchi K. Isolation of extrachromosomal deoxyribonucleic acids from extremely thermophilic bacteria. J Gen Microbiol. 1978 Feb;104(2):193–199. doi: 10.1099/00221287-104-2-193. [DOI] [PubMed] [Google Scholar]
- Jennings S. M., Tsay Y. H., Fisch T. M., Robinson G. W. Molecular cloning and characterization of the yeast gene for squalene synthetase. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6038–6042. doi: 10.1073/pnas.88.14.6038. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Koyama Y., Hoshino T., Tomizuka N., Furukawa K. Genetic transformation of the extreme thermophile Thermus thermophilus and of other Thermus spp. J Bacteriol. 1986 Apr;166(1):338–340. doi: 10.1128/jb.166.1.338-340.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Liao H., McKenzie T., Hageman R. Isolation of a thermostable enzyme variant by cloning and selection in a thermophile. Proc Natl Acad Sci U S A. 1986 Feb;83(3):576–580. doi: 10.1073/pnas.83.3.576. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Misawa N., Nakagawa M., Kobayashi K., Yamano S., Izawa Y., Nakamura K., Harashima K. Elucidation of the Erwinia uredovora carotenoid biosynthetic pathway by functional analysis of gene products expressed in Escherichia coli. J Bacteriol. 1990 Dec;172(12):6704–6712. doi: 10.1128/jb.172.12.6704-6712.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miyazaki K., Eguchi H., Yamagishi A., Wakagi T., Oshima T. Molecular cloning of the isocitrate dehydrogenase gene of an extreme thermophile, Thermus thermophilus HB8. Appl Environ Microbiol. 1992 Jan;58(1):93–98. doi: 10.1128/aem.58.1.93-98.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ray P. H., White D. C., Brock T. D. Effect of growth temperature on the lipid composition of Thermus aquaticus. J Bacteriol. 1971 Oct;108(1):227–235. doi: 10.1128/jb.108.1.227-235.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sandmann G., Misawa N. New functional assignment of the carotenogenic genes crtB and crtE with constructs of these genes from Erwinia species. FEMS Microbiol Lett. 1992 Jan 15;69(3):253–257. doi: 10.1016/0378-1097(92)90656-9. [DOI] [PubMed] [Google Scholar]
- Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ulrich J. T., McFeters G. A., Temple K. L. Induction and characterization of -galactosidase in an extreme thermophile. J Bacteriol. 1972 May;110(2):691–698. doi: 10.1128/jb.110.2.691-698.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]