Department of Chemistry and Chemical Biology, Indiana University-Purdue University Indianapolis, 402 N Blackford St, Indianapolis, IN, 46202, USA.
Department of Chemistry and Biochemistry, Duquesne University, Pittsburgh, PA, USA.
J Biol Inorg Chem. 2020 Jun;25(4):547-569. doi: 10.1007/s00775-020-01787-y. Epub 2020 Apr 11.
Mononuclear molybdenum enzymes catalyze a variety of reactions that are essential in the cycling of nitrogen, carbon, arsenic, and sulfur. For decades, the structure and function of these crucial enzymes have been investigated to develop a fundamental knowledge for this vast family of enzymes and the chemistries they carry out. Therefore, obtaining abundant quantities of active enzyme is necessary for exploring this family's biochemical capability. This mini-review summarizes the methods for overexpressing mononuclear molybdenum enzymes in the context of the challenges encountered in the process. Effective methods for molybdenum cofactor synthesis and incorporation, optimization of expression conditions, improving isolation of active vs. inactive enzyme, incorporation of additional prosthetic groups, and inclusion of redox enzyme maturation protein chaperones are discussed in relation to the current molybdenum enzyme literature. This article summarizes the heterologous and homologous expression studies providing underlying patterns and potential future directions.
单核钼酶催化多种反应，这些反应在氮、碳、砷和硫的循环中至关重要。几十年来，为了对这一庞大的酶家族及其所进行的化学反应有一个基本的了解，人们一直在研究这些关键酶的结构和功能。因此，获得大量具有活性的酶对于探索这个家族的生化能力是必要的。这篇综述总结了在表达单核钼酶过程中遇到的挑战，并提出了一些方法。本文还讨论了钼辅因子合成和掺入、表达条件的优化、活性与非活性酶的分离、额外辅基的掺入以及包含氧化还原酶成熟蛋白伴侣等方面的有效方法，这些方法都与当前的钼酶文献有关。本文总结了异源和同源表达研究，提供了潜在的未来方向。
