2007 Fiscal Year Final Research Report Summary
Molecular mechanism of multiple dissimilatory nitrogen metabolisms by eukaryotic and prokaryotic cells
| Project/Area Number |
14104005
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (S)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
応用微生物学・応用生物化学
|
|---|
| Research Institution | The University of Tokyo |
|---|
Principal Investigator |
SHOUN Hirofumi The University of Tokyo, GRADUATE SCHOOL OF AGRICULTURAL AND LIFE SCIENCES, PROFESSOR (70012036)
|
|---|
| Project Period (FY) |
2002 – 2007
|
| Keywords | fungal denitrification / copper-containing nitrite reducatse / cytochrome P450nor / ammonia fermentation / Fusarium oxysporum / codenitrification / anaerobic respiration by actinomycetes / aerobic denitrifying bacteria |
|---|
| Research Abstract |
(1) Molecular mechanisms of fungal denitrification system: The genes encoding copper-containing nitrite reductase (nirK) were isolated from several fungi including Fusarium oxysporum, in which fungal denitrifying activity was first found. The NirK protein previously isolated from F. oxysporum, was shown to be the product of the nirK gene. This is the first isolation from eukaryote of an orthologoue of the bacterial counterpart involved in denitrification. Phylogenetic analysis shows that these nirK genes and their homologue genes that have been recently found in many eukaryotic microorganisms are originated from the single ancestor, suggesting that they are derived from the protomitochondrion. It was shown that the nitrate reducing system of F. oxysporum comprises nitrate reductase (dNaR, possibly NarGHI, which is universally found in bacterial nitrate respiration) and ubiquinone-dependent formate dehydrogenase. Unlike the bacterial system the fungal dNaR system was shown to function s
… More
imultaneously with cytochrome oxidase under micro-aerobic conditions, which we termed hybrid respiration. These results have revealed general occurrence of fungal denitrification in nature.
(2) Reaction mechanism of cytochrome P450nor: P450nor participates in fungal denitrification as nitric oxide (NO) reductase, performing an unusual electron transfer, direct transfer of 2 electrons from NADH to the hemeprotein. Structural and kinetic analyses such as determination of the crystal structure of P450nor in complex with an NAD analogue revealed-the detailed reaction mechanism of P450nor, including the mechanism of the prochiral hydride transfer from NADH to the ferric heme-NO complex and identification of the chemical entity of the key spectral intermediate. The results should have gave an marked impact, to many field such as bioinorganic chemistry and P450 biochemistry.
(3) Ammonia fermentation and anaerobic growth of fungi: Involvement of the assimilatory nitrate reducing system in the fungal ammonia fermentation was confirmed by characterizing the uiaD or niiA-defective mutant of the fungus Aspergillus nidulans. The denitrifying fungus F. oxysporum was shown to perform hetero-lactic acid fermentation, which is coupled to ammonia fermentation. The results together with those of denitrification show that al least a portion of soil fungi are facultative with respect to aeration, not obligate aerobe as previously thought.
(4) Other subjects: Codenitrification was shown to be a biological reaction but not chemical reaction, which was supported by the results of its saturation kinetics against codenitrification substrates, and inhibition by respiration inhibitors such as cyanide. Actinomycetes are thought to be obligate aerobes, By contrast, we provided evidence for occurrence of nitrate respiration in the actinomycetes Streptomyces antibioticus and Streptomyces coelicolor. Aerobic denitrifying bacteria were isolated in order to apply them to the control of green house-effect gas emission, and their properties were characterized. Less
|
Research Products
(91 results)
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
[Journal Article] Structural evidence for direct hydride transfer from NADH to cytochrome P450nor.2004
Author(s)
Oshima, R. . Fushinobu, S., Su, F., Zhang, L., Takaya, N., Shoun, H
-
Journal Title
J. Mol. Biol 342
Pages: 207-217
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
[Journal Article] Aerobic denitrifying bacteria that produce low levels of nitrous oxide.2003
Author(s)
Takaya, N., Catalan, Sakairi, M. A. B., Sakaguchi, Y., Kato, I., Zhou, Z., Shoun, H
-
Journal Title
Appl. Environ. Microbiol. . 69(6)
Pages: 3152-3157
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
-
-
-
-
[Journal Article] Hybrid respiration in the denitrifying mitochondria of Fusarium oxysporum. .2003
Author(s)
Takaya, N., Kuwazaki, S., Adachi, Y., Suzuki, S., Kikuchi, T., Nakamura, H., Shiro, Y., Shoun H.
-
Journal Title
J. Biochem. 133
Pages: 461-465
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
-
-
-
[Journal Article] Aerobic denitrifying bacteria that produce low levels of nitrous oxide.2003
Author(s)
Takaya, N., Catalan-Sakairi, M. A. B., Sakaguchi, Y., Kato, I., Zhou, Z., Shoun, H.
-
Journal Title
Appl. Environ. Microbiol. 69(6)
Pages: 3152-315
Description
「研究成果報告書概要(欧文)」より
-
-
-
-
[Journal Article] Hybrid respiration in the denitrifying mitochondria of Fusarium oxysporum.2003
Author(s)
Takaya, N., Kuwazaki, S., Adachi, Y., Suzuki, S., Kikuchi, T., Nakamura, H., Shiro, Y., Shoun H.
-
Journal Title
J. Biochem. 133
Pages: 461-465
Description
「研究成果報告書概要(欧文)」より
-
-
-
-
-
-
-
-
-
[Journal Article] Copper-containing nitrite reductase gene of the fungus Aspergillus oryzae.
Author(s)
Nakanishi, Y, Zhou, S, Kim, S-W, Fushinobu, S, Wakagi, T, Shoun, H
-
Journal Title
Description
「研究成果報告書概要(欧文)」より
-
-
-
-
[Journal Article] Codenitrification by the fungus Fusarium solani.
Author(s)
Jian, R, Kim, D-H, Takaya, N, Fushinobu, S, Wakagi, T, Shoun, H
-
Journal Title
Description
「研究成果報告書概要(欧文)」より
-
-
[Journal Article] NADH-peroxidase activity catalyzed by cytochrome P450nor.
Author(s)
Kurose, T, Ozaki, M, Nakaya, S, Fushinobu, S, Wakagi, T, Shoun, H
-
Journal Title
Description
「研究成果報告書概要(欧文)」より
-
-
-
-
-
-
-
-
-
-
[Presentation] 酵母の脱窒系の解明2006
Author(s)
太田匠吾, 高木恒輝, 伏信進矢, 若木高善, 祥雲弘文
Organizer
日本農芸化学会2006年度大会
Place of Presentation
京都
Year and Date
20060325-20060328
Description
「研究成果報告書概要(和文)」より
-
-
-
-
-
-
-
[Presentation] 放線菌の新規窒素代謝の解析2003
Author(s)
森田綾子, 青木玲二, 佐々木康幸, 高谷直樹, 伏信進矢, 若木高善, 祥雲弘文
Organizer
日本農芸化学会2003年度大会
Place of Presentation
東京
Year and Date
20030331-20030403
Description
「研究成果報告書概要(和文)」より
-
-
-
[Presentation] P450norの共脱窒活性2003
Author(s)
蘇霏, 高谷直樹, 祥雲弘文
Organizer
日本農芸化学会2003年度大会
Place of Presentation
東京
Year and Date
20030331-20030403
Description
「研究成果報告書概要(和文)」より
-
-
-
-
-
-
-
-
