Abstract

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.

Graphic abstract

中文翻译：

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功能性单核钼酶：在分子克隆，表达和分离中的挑战和胜利。

摘要

单核钼酶催化氮，碳，砷和硫循环过程中必不可少的各种反应。数十年来，已经对这些关键酶的结构和功能进行了研究，以开发出这一庞大的酶家族及其所执行的化学方法的基础知识。因此，获得大量活性酶对于探索该家族的生化能力是必要的。这篇小型综述总结了在过程中遇到的挑战中过表达单核钼酶的方法。钼辅因子合成和掺入的有效方法，优化表达条件，改善活性与非活性酶的分离，掺入其他辅基，关于钼酶的文献，讨论了氧化还原酶成熟蛋白伴侣蛋白的组成和组成。本文总结了异源和同源表达研究，提供了潜在的模式和潜在的未来方向。

图形摘要