Characterization of the enzyme system catalyzing Anammox

催化厌氧氨氧化酶系统的表征

• 批准号: 13650861
• 负责人: FUJII Takao
• 金额: $ 1.86万
• 依托单位: Sojo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2001
• 资助国家: 日本
• 起止时间: 2001 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13650861/
• 关键词: anaerobic ammonium oxidation; Anammox; Planctomycete; denitrification; treatment of nitrite and ammonium; Planctomycete; FISH

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.

我们的实验室正在进行熊本地下水处理的厌氧氨氧化（Anammox）生物膜培养物的富集和表征。为了研究成熟生物膜群落的细菌组成，通过PCR扩增16 S rDNA序列，并使用DNA数据库进行比较分析。只有一个序列与第一个发现的厌氧氨氧化浮游菌-荧光成像显示KSU-1形成球形簇，包裹在一层薄薄的动胶菌中。我们试图纯化羟胺氧化还原酶（hao），它似乎是厌氧氨氧化的关键酶。从富集的厌氧氨氧化生物膜中提取hao，用DEAE-纤维素Superdex凝胶过滤柱层析纯化，并用纤维素膜透析，通过小于50 kDa的分子。SDS-PAGE分析表明，纯化后的hao具有均一性，其分子量约为110 kDa。虽然hao的光谱与欧洲亚硝化单胞菌相似，但肼对hao的光谱变化大于羟胺，表明hao对肼的亲和力高于羟胺。

项目成果

U.Imajo, H.Ishida, T.Fujii, H.Sugino, J.Rouse, K.Furukawa: "Detection of anammox activity from activated sludges"Asian water qual 2001, Asia-pacific regional conference procceding. (CD-ROM). (2001)

U.Imajo、H.Ishida、T.Fujii、H.Sugino、J.Rouse、K.Furukawa：“活性污泥中厌氧氨氧化活性的检测”2001 年亚洲水质量，亚太地区会议记录。

T.Fujii, H.Sugino, J.D.Rouse, K.Furukawa: "Characterization of the microbial community in an anaerobic ammonium-oxidizing biofilm cultured on a nonwoven biomass carrier"Journal of Bioscience and Bioengineering. 94. 412-418 (2002)

T.Fujii、H.Sugino、J.D.Rouse、K.Furukawa：“在非织造生物质载体上培养的厌氧铵氧化生物膜中微生物群落的特征”生物科学与生物工程杂志。

U. Imajo: "Detection of anammox activity from activated sludges"Asian water qual 2001, Asia-pacific regional conference proceeding, CD-ROM. (2001)

U. Imajo：“活性污泥中厌氧氨氧化活性的检测”，2001 年亚洲水质量，亚太地区会议论文集，CD-ROM。

K. Furukawa: "Establishment of an aerobic ammonium-oxidizing culture in continuous treatment with non-woven biomass carrier"Proceeding of Water environment federation, 74th annual conference and exposition, CD-ROM. (2001)

K. Furukawa：“用无纺生物质载体连续处理好氧氨氧化培养物”，水环境联合会论文集，第 74 届年会暨博览会，CD-ROM。

U.Imajo, H.Ishida, T.Fujii, H.Sugino, J.Rouse, K.Furukawa: "Detection of anammox activity from activated sludges"Asian water qual 2001, Asia-pacific regional conference proceeding. (CD-ROM). (2001)

U.Imajo、H.Ishida、T.Fujii、H.Sugino、J.Rouse、K.Furukawa：“活性污泥中厌氧氨氧化活性的检测”2001 年亚洲水质量，亚太地区会议论文集。