| Co-Investigator(Kenkyū-buntansha) |
ISHINO Yoshizumi Takara Shuzo, Biotechnology Res.Lab., Senior Researcher, 中央研究所, 主任研究員
OWADA Koichi Univ.Tokyo, Inst.Oceanography, Professor, 海洋研究所, 教授 (30013585)
SAKO Yoshihiko Kyoto Univ., Agirulture, Lecturer, 農学部, 講師 (60153970)
UCHIDA Aritsune Kyoto Univ., Agriculture, Associate Prpfessor, 農学部, 助教授 (50027190)
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| Research Abstract |
Recent isolations and descriptions of new microorganisms from the hydrothermal vents(>200ﾟC)have been of ecological, physiological, and biotechnological interest, especially with respect of hyperthermophile. Its has not made use of these hyperthermohpiles because of difficulty in cultivation under hightemperature and pressure. In this study, we firstly establish the isolation and cultivation system of marine hyperthemophiles growing at 80 to 100ﾟC which are available for biotechnology and then attempt to use heat stable enzymes for biotechnological purpose. The results are as follows ;
(1)The anaerobic cultivation system is developed by use of heat and high pressure tolerant glass bottlea and butyl rubber stoppers containing a gas mixture of 80%H_2 and 20%CO_2 at 0.2MPa. For culturing aerobic organisms at temperature up to 95ﾟC the new gyrotary shaker is developed by composing of heat torelant grease and super insulating materials.
(2)We isolated new anaerobic hypethermophiles Pyrococcus TS100, Methanococcus TS300, aerobic Protothermus maritima OKD7 from shallow hydrothermal vents in Tachibana bay, Nagasaki and the novel marine hyperthermophilic Archaon Aeropyrus pernix Kl from Kodakara island, Kagoshima. Optimum temperature of TS100, TS300, OKD7and Kl are 103,88,80and90-95ﾟC,respectively. The sequences of 16SrDNA shows that A.pernix Kl in a primitive Crenarchaeta and P.maritima OKD7 is a primive the Bacteria.
(3)Nonallosteric CO_2 fixing enzyme (phosphoenolpyruvate carboxylase) was purified and characterized from methanogenic bacterium TS300, therefore it might be useful for diagnosis of cardiac insufficiency. A.pernix Kl produces a useful extracellular metalloproteinase which shows optimum temperature of 110ﾟC and theextreme themostability (half life at 120ﾟC,150min). DNA polymerase of Pyrococcus TS100 is a very thermostable and useful for PCR method.
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