HYPOTHETICAL PROTEIN FROM PODOSPORA ANSERINA AS A NITROALKANE OXIDASE

该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。次级项目的主要支助 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 为子项目列出的总成本可能 代表子项目使用的中心基础设施的估计数量， NCRR赠款不直接向子项目或子项目工作人员提供资金。 尖孢镰刀菌（Fusarium oxysporum）的黄素蛋白硝基烷氧化酶（NAO）催化伯硝基烷和仲硝基烷氧化成它们各自的醛和酮。除F.尖孢霉的DNA序列已被注释为NAO，因此进行可用序列数据库的搜索以鉴定其他潜在的NAO。根据F.尖孢序PODANSg 2158作为重组酶过表达并分离至高纯度，使其适合于动力学研究和结晶试验。

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.