2002 Fiscal Year Final Research Report Summary
Characterization of the enzyme system catalyzing Anammox
| Project/Area Number |
13650861
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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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 |
FUJII Takao Sojo university, Department of Engineering, Professor, 工学部, 教授 (80165331)
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| Co-Investigator(Kenkyū-buntansha) |
SUGINO Hiroyuki Sojo university, Department of Engineering, 工学部, Lecturer (90258434)
FURUKAWA Kenji Kumamoto university, Department of Engineering, Professor, 工学部, 教授 (60029296)
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| Project Period (FY) |
2001 – 2002
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| Keywords | anaerobic ammonium oxidation / Anammox / Planctomycete / denitrification / treatment of nitrite and ammonium |
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| Research Abstract |
The enrichment and characterization of anaerobic ammonium-oxidizing (Anammox) biofilm cultures for the treatment of groundwater in Kumamoto, are ongoing in our laboratories. To investigate the bacterial composition of the mature biofilm community, 16S rDNA sequences were amplified by PCR and comparative analyses using DNA databases were conducted. Only one sequence had a notable similarity (92.2%) to that of the first discovered anaerobic ammonium-oxidizing planctomycete, Candidatus Brocadia anammoxidans, The newly discovered strain (designated KSU-1) was dominant among detectable members of the biofilm community. By fluorescence imaging, KSU-1 was shown to form spherical clusters wrapped in a thin layer of Zoogloea sp.
We attempted to purify the hydroxylamine oxidoreductase(hao) seemed as a critical enzyme in Anammox. The hao was extracted from the enriched Anammox biofilm, purified with DEAE-cellulose Superdex gel filtration column chromatography, and dialyzed with a cellulose membrane passing less than 50 kDa molecules. The purified hao was homogeneous by SDS-PAGE, exhibiting a 110 kDa band smaller than that of hao purified from Candidatus Brocadia anammoxidans. Although the hao showed similar spectra to that from Nitrosomonas europeae, hydrazine gave lager spectral change of the hao than hydroxylamine, suggesting it has higher affinity to hydrazine than hydroxylamine.
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