2000 Fiscal Year Final Research Report Summary
Studies on the application of the gene of oligotrophic bacteria
| Project/Area Number |
10450314
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| Research Category |
Grant-in-Aid for Scientific Research (B).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
生物・生体工学
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| Research Institution | Sojo University |
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Principal Investigator |
OGAWA Takahira Sojo University, Faculty of Engineering, Professor, 工学部, 教授 (40029244)
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| Co-Investigator(Kenkyū-buntansha) |
MASTUOKA Masayoshi Sojo University, Faculty of Engineering, Associate Professor, 工学部, 助教授 (10121667)
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| Project Period (FY) |
1998 – 2000
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| Keywords | oligotrophic bacteria / organotin / triphenyltin / diphenyltin / isocitrate lyase / isocitrate lyase gene |
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| Research Abstract |
1. Objective Oligotrophic bacteria have not been studied yet in the biotechnology fields. This project was to search for the applicable genes from oligotrophic bacteria. We firstly tried to isolate the organotins (especially triphenyltin) degradation oligotrophs, and secondly sequence the isocitrate lyase gene from an oligotrophic bacterium.
2. Methods Authentic triphenyltin cholide and its derivatives were shaken with 2-hydroxy-2,4,6-cycloheptatrienone (tropolone) and extracts were treated with Grihnard reagent (propyl magnesium chloride) and asssayed by gas chromatography-FPD.Triphenyltin degradation microbes were screened from triphenyltin resistant oligotrophic bacteria which were isoltated from the sea and soils. The isolation methods and its DNA sequence of isocitrate lyase gene from oligotrophic bacteria were used standard protocols.
3. Results and Discussion (1) Triphenyltin and diphynyltin could be separated and detected by GC-FPD.(2) Acclimated oligotrophic bacterial population could degrate triphenyltin, but pure oligotrophic strains could not degrate triphenyltin.
(3) Many oligotrophic bacteria were isolated from natural sources, and we isolated the isocitrate lyase (ICL) gene from a pure oligotrophic bacterium named OT622. The enzymatic properties and the DNA sequence of the ICL gene were studied. The Km value for isocitrate of the isocitrate lyase had 0.075mM and this value was very small (ca.1/3) compared with that of Yarrowia lipolytica. This means genes of oligotrophs are useful for the biotechnology fields.
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