Abstract

NADPH-dependent oxidoreductases are crucial biocatalysts for the industrial production of chiral compounds. For in situ recycling of required expensive cofactors in this biosynthetic process, NAD(P)⁺-dependent formate dehydrogenases (FDHs) are wanted to be employed as redox biocatalysts due to their greener and process friendly nature. However, their utilization is limited by their undesired cofactor preference that strongly prone to NAD⁺ more than NADP⁺ and catalytic efficiency. To mine NADP⁺-dependent FDHs in nature by the guidance of bioinformatic tools or re-engineering of their NAD⁺-dependent equivalents to get an applicable recycler are attractive topics in bioengineering. It can be said that, up to now, the attempts to switch the cofactor preference of the FDHs generally have resulted in NADP⁺-dependent enzymes that have not to catch the desired catalytic efficiencies or stability. In this review, all studies about the native NADP⁺-dependent FDHs and also engineered equivalents that reconstructed with different protein engineering approaches for altering the coenzyme specificity are outlined. To switch the coenzyme preference of FDHs or to find the native NADP⁺-dependent FDHs will be the hot topics in bioengineering until finding a feasible regenerator. Therefore, this study will be a useful guide to get a pathway for designing or discovering novel NADP⁺-dependent FDHs.

摘要

NADPH 依赖性氧化还原酶是手性化合物工业生产的关键生物催化剂。为了在该生物合成过程中原位回收所需的昂贵辅因子，NAD(P) ⁺依赖性甲酸脱氢酶 (FDH) 由于其更环保和工艺友好的性质而被希望用作氧化还原生物催化剂。然而，它们的利用受到它们不希望的辅助因子偏好的限制，这些辅助因子偏好强烈倾向于 NAD ⁺多于 NADP ⁺和催化效率。通过生物信息学工具的指导或对其 NAD ^{+ 的}重新设计，在自然界中挖掘 NADP ⁺依赖性 FDH获得适用的回收器的依赖等价物是生物工程中有吸引力的主题。可以说，到目前为止，改变 FDH 的辅因子偏好的尝试通常导致 NADP ⁺依赖性酶无法获得所需的催化效率或稳定性。在这篇综述中，概述了所有关于天然 NADP ⁺依赖性 FDH 的研究，以及用不同的蛋白质工程方法重建以改变辅酶特异性的工程等效物。切换 FDH 的辅酶偏好或寻找天然 NADP ⁺依赖 FDHs 将成为生物工程的热门话题，直到找到可行的再生器。因此，这项研究将是获得设计或发现新型 NADP ⁺依赖性 FDH途径的有用指南。