HYPOTHETICAL PROTEIN FROM PODOSPORA ANSERINA AS A NITROALKANE OXIDASE

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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.