2001 Fiscal Year Final Research Report Summary
Genetic analysis of bacterial formaldehyde-fixing enzyme system and tis application of useful compound production
| Project/Area Number |
11460041
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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 |
応用微生物学・応用生物化学
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
KATO Nobuo Graduate School of Agriculture Division of Applied Life Sciences, Kyoto University Professor, 農学研究科, 教授 (50026556)
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| Co-Investigator(Kenkyū-buntansha) |
YURIMOTO Hiroya Graduate School of Agriculture Division of Applied Life Sciences, Kyoto University, Assistant Professor, 農学研究科, 助手 (00283648)
SAKAI Yasuyoshi Graduate School of Agriculture Division of Applied Life Sciences, Kyoto University, Associate Professor, 農学研究科, 助教授 (60202082)
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| Project Period (FY) |
1999 – 2001
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| Keywords | Bacillus subtilis / Mycobacterium gastri / Methylomonas aminofaciens / Pyrococcus horikoshii / 3-Hexulose-6-phosphate synthase / 6-Phospho-3-hexuloisomerase / Fonnaldehyde-fixing enzyme system / Ribulose monophosphate pathway |
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| Research Abstract |
1) Genetic analysis of bacterial formaldehyde-fixing enzyme system : The genes for formaldehydefixing enzyme system in the ribulose monophosphate pathway, involving 3-hexulose-6-phosphate synthase (EPS) and 6-phospho-3-hexuloisomerase (PHI), were cloned from the facultative methylotroph, Mycobacterium gastri MB 19, and the obligate methylotroph, Methylomonas aminofaciens 77a. The two strain were different in the gene organization and regulation mechanism for the gene expression. The homologous proteins to HPS and PHI are found in a variety of strains of Archaea and Bacteria.
2) Purification of HPS and PHI from a methylotrophic and thermophilic bacterium, Bacillus brevis, and their gene analyses : HPS and PHI were purified from B. brevis and characterized A DNA fragment containing hpi and phi was cloned. From the genetic analysis for the fragment, gene cluster is an operon for hps/phi, whose expression occurred only the presence of methanol or formaldehyde.
3) Regulation in the expression of formaldehyde-fixing (hps/phi) operon in Bacillus subtilist : The gene product of yckH, which locates upstream of hps/phi operon, was purified and characterized. Several nucleotide sequence on which yckH bound specifically were found. From these and another results, it is concluded that YckH is essential for the transcription of hps/phi operon.
4) Expression and purification of HPS and PHI from a hyper thermophilic Archaean, Pyrococcus horikoshii : DNA fragments corresponding hps and phi were constructed from a DNA fragment (PH1938) of P. horikoshii. It has been confirmed that the two DNA fragments and PH1938 are enough to expression of the active enzymes for formaldehyde fixation. This is the first evidence that genes for formaldehyde fixation are active in an Archaeon.
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Research Products
(11 results)
