RUI: Characterization of the Manganese Centers of Oxalate Oxidase from Ceriporiopsis subvermispora

本项目旨在阐明蛋白质环境如何调节重组Ceriporiopsis subvermispora草酸氧化酶的草酸降解化学。 草酸氧化酶催化草酸盐的碳-碳键断裂以产生二氧化碳和过氧化氢。 虽然目前还没有C. subvermispora（CsOxOx），序列数据和同源性建模表明，它是第一个含锰的bicupin酶鉴定催化该反应。 特征最好的草酸氧化酶来自大麦和小麦。 这些酶，也被称为germins，含有一个单一的cupin结构域，因此被归类为monocupin。 有趣的是，CsOxOx与bicupin微生物草酸脱羧酶具有最大的序列同源性。 本研究的近期目标是：1）研究C. subvermispora和2）鉴定酶的活性位点。 这项研究有望有助于理解微妙的结构变化如何影响进化相关蛋白质的显着功能变异。 表征CsOxOx的锰依赖性是重要的，以便将这种酶置于其他草酸盐降解酶和其他cupin蛋白的背景下。 确定CsOxOx的活性位点是一个重要的问题。 如果只有一个锰中心介导催化，提出了关于第二个锰结合结构域的功能（如果有的话）以及每个结构域中局部蛋白质结构导致差异反应性的程度的关键问题。（格鲁吉亚大学系统的两年制单位）从一个提供优秀科学课程的机构转变为一个也为学生提供参与有意义和重要的实验室研究的机会的机构。 本科生在盖恩斯维尔州立学院（GSC）开展大部分工作，但也有机会与佛罗里达大学和佛罗里达州塔拉哈西国家高磁场实验室的合作者一起工作。学生研究人员有望通过多种媒体向广大受众传达他们的研究经验。 学生研究人员还应在专业会议上和区域教育界发表他们的研究结果和报告研究结果。 该项目的资金用于购买，运营和维护GSC无法获得的研究设备。

This project seeks to elucidate how protein environment modulates the chemistry of oxalate degradation in recombinant oxalate oxidase from Ceriporiopsis subvermispora. Oxalate oxidase catalyzes the carbon-carbon bond cleavage of oxalate to yield carbon dioxide and hydrogen peroxide. Although there is currently no structural information available for oxalate oxidase from C. subvermispora (CsOxOx), sequence data and homology modeling indicate that it is the first manganese-containing bicupin enzyme identified that catalyzes this reaction. The best characterized oxalate oxidases are from barley and wheat. These enzymes, also known as germins, contain a single cupin domain and are therefore classified as monocupins. Interestingly, CsOxOx shares greatest sequence homology with the bicupin microbial oxalate decarboxylases. The short term goals of this research are to 1) to characterize the manganese-dependence of oxalate oxidase from C. subvermispora and 2) to identify the active site of the enzyme. This research is expected to contribute to the understanding of how subtle structural changes effect remarkable functional variation in evolutionarily related proteins. Characterizing the manganese-dependence of CsOxOx is significant in order to place this enzyme in the context of other oxalate degrading enzymes and that of other cupin proteins. Identifying the active site of CsOxOx is an important problem. If only one of the manganese centers mediates catalysis, critical questions are raised concerning the function (if any) of the second Mn-binding domain and the extent to which local protein structure in each domain results in differential reactivity.Broader ImpactsThis project is transforming Gainesville State College (a two-year unit of the University System of Georgia) from an institution that provides excellent coursework in the sciences into one that also provides students the opportunity to participate in meaningful and important laboratory research. Undergraduate students carry out most of this work at Gainesville State College (GSC), but also have the opportunity to work with collaborators at the University of Florida and the National High Magnetic Field Laboratory in Tallahassee, FL. Student researchers are expected to communicate their research experiences to broad audiences through diverse media. Student researchers also are expected to publish their results and report research findings at professional meetings and to the regional educational community. Funds from this project are used to acquire, operate, and maintain research equipment that is not otherwise available at GSC.