HYPOTHETICAL PROTEIN FROM PODOSPORA ANSERINA AS A NITROALKANE OXIDASE

来自雁丝足孢菌的假想蛋白质作为硝基烷氧化酶

• 批准号: 8361709
• 负责人: PAUL F. FITZPATRICK
• 金额: $ 1.1万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361709
• 关键词: 2-Nitropropane dioxygenase; Active Sites; Aldehydes; Crystallization; Databases; Enzymes; Flavoproteins; Funding; Fusarium; Grant; Ketones; Kinetics; National Center for Research Resources; PAWR protein; Podospora anserina; Principal Investigator; Recombinants; Research; Research Infrastructure; Resources; Source; United States National Institutes of Health; cost; nitroalkane; overexpression; oxidation; structural biology

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The flavoprotein nitroalkane oxidase (NAO) from Fusarium oxysporum catalyzes the oxidation of primary and secondary nitroalkanes to their respective aldehydes and ketones. No enzymes other than that from F. oxysporum have been annotated as NAOs so a search of available sequence databases was performed to identify other potential NAOs. Several fungal enzymes including the hypothetical protein PODANSg2158 from Podospora anserina were identified with conserved active site residues D402, R409, and S276 according to the F. oxysporum sequence. PODANSg2158 was overexpressed as a recombinant enzyme and isolated to high purity making it suitable for kinetic studies and crystallization trials.

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 尖孢镰刀菌中的黄素蛋白硝基烷烃氧化酶(NAO)能催化初级和次级硝基烷烃氧化成相应的醛和酮。除了来自尖孢镰刀菌的酶外，没有其他酶被注释为NaOS，因此对现有的序列数据库进行了搜索，以确定其他潜在的NaOS。根据尖孢霉菌的序列，鉴定了几种真菌酶，其中包括来自鹅座孢霉的假想蛋白PODANSg2158，其中含有保守的活性位点残基d402、R409和S276。PODANSg2158是一种高效表达的重组酶，其纯度较高，适合于动力学研究和结晶实验。