Abstract
We isolated a highly transformable thermophile, Bacillus stearothermophilus SIC1, which exhibited the following features. The growth temperature ranged from 45 to 65°C in L broth. The maximum cell concentration in 2L broth (2% tryptone, 1% yeast extract, 0.5% NaCl, pH 7.2) was determined as an optical density at 660 nm of 7.8, and the generation time was 11 min at 60°C. Strain SIC1 was a prototroph and was transformed by the protoplast procedure not only with repB plasmids (high-copy-number plasmids such as pTB913 and pUB110) but also with repA plasmids (low-copy-number plasmids such as pTB53). Transformation efficiencies with repB and repA plasmids were about 2 × 106 to 5 × 106 and 5 × 104 transformants per μg of DNA, respectively. The transformant carrying plasmid pTB913Y/K could grow at 63°C in the presence of kanamycin. The regeneration frequency of protoplasts was 60%, and only 1 day was needed for regeneration at 55°C.
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- Aiba S., Kitai K., Imanaka T. Cloning and Expression of Thermostable alpha-Amylase Gene from Bacillus stearothermophilus in Bacillus stearothermophilus and Bacillus subtilis. Appl Environ Microbiol. 1983 Nov;46(5):1059–1065. doi: 10.1128/aem.46.5.1059-1065.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen Z. F., Wojcik S. F., Welker N. E. Genetic analysis of Bacillus stearothermophilus by protoplast fusion. J Bacteriol. 1986 Mar;165(3):994–1001. doi: 10.1128/jb.165.3.994-1001.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fujii M., Takagi M., Imanaka T., Aiba S. Molecular cloning of a thermostable neutral protease gene from Bacillus stearothermophilus in a vector plasmid and its expression in Bacillus stearothermophilus and Bacillus subtilis. J Bacteriol. 1983 May;154(2):831–837. doi: 10.1128/jb.154.2.831-837.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Imanaka T., Ano T., Fujii M., Aiba S. Two replication determinants of an antibiotic-resistance plasmid, pTB19, from a thermophilic bacillus. J Gen Microbiol. 1984 Jun;130(6):1399–1408. doi: 10.1099/00221287-130-6-1399. [DOI] [PubMed] [Google Scholar]
- Imanaka T., Fujii M., Aramori I., Aiba S. Transformation of Bacillus stearothermophilus with plasmid DNA and characterization of shuttle vector plasmids between Bacillus stearothermophilus and Bacillus subtilis. J Bacteriol. 1982 Mar;149(3):824–830. doi: 10.1128/jb.149.3.824-830.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kubo M., Imanaka T. Cloning and nucleotide sequence of the highly thermostable neutral protease gene from Bacillus stearothermophilus. J Gen Microbiol. 1988 Jul;134(7):1883–1892. doi: 10.1099/00221287-134-7-1883. [DOI] [PubMed] [Google Scholar]
- Ljungdahl L. G. Physiology of thermophilic bacteria. Adv Microb Physiol. 1979;19:149–243. doi: 10.1016/s0065-2911(08)60199-x. [DOI] [PubMed] [Google Scholar]
- Matsumura M., Katakura Y., Imanaka T., Aiba S. Enzymatic and nucleotide sequence studies of a kanamycin-inactivating enzyme encoded by a plasmid from thermophilic bacilli in comparison with that encoded by plasmid pUB110. J Bacteriol. 1984 Oct;160(1):413–420. doi: 10.1128/jb.160.1.413-420.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Matsumura M., Yasumura S., Aiba S. Cumulative effect of intragenic amino-acid replacements on the thermostability of a protein. 1986 Sep 25-Oct 1Nature. 323(6086):356–358. doi: 10.1038/323356a0. [DOI] [PubMed] [Google Scholar]
- Nakajima R., Imanaka T., Aiba S. Nucleotide sequence of the Bacillus stearothermophilus alpha-amylase gene. J Bacteriol. 1985 Jul;163(1):401–406. doi: 10.1128/jb.163.1.401-406.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]