Abstract: Cofactor regeneration and repeated utilization showed important effect on the preparation of chiral compound by the redox biocatalysts. Cofactor regenerated by enzymatic catalysis had high efficiency and specificity，such cofactor regenerated system include single cell in vivo cofactor regenerated system，double cell coupling system and in vitro enzyme coupling system due to their catalysis circumstance. The construct methods of different cofactor regenerated system were presented particularly in this paper. The possible issues in practical application of these cofactor regenerated system were especially pointed out and the resolve methods were afforded，such as the solution of low water-soluble organic substrate by ionic liquid，the subsistence of micro-molecule cofactor by covalent immobilization and the utilization of endogenous cofactor regenerated enzyme in cell free extract. The advantage and limitation of different cofactor regenerated systems that applied in industry catalysis were also commented，more attention should be focused on the repeated and multi-batches utilization of the cofactor regenerated system.

Key words: chiral compound, cofactor regenerated system, enzyme coupling reaction, cell coupling, endogenous cofactor regenerated enzyme

摘要： 辅酶的再生与反复利用对氧化还原型生物催化剂在手性化合物中的生产具有重要作用。辅酶的酶催化法再生具有高效、专一的特点，根据其催化环境可分为单细胞内辅酶再生体系、双细胞耦合体系及胞外酶耦合体系。本文详细阐述了酶法再生中各种辅酶再生体系的构建方法，特别指出了辅酶再生体系使用中可能遇到的问题并给出了解决方法，如利用离子液体解决有机底物低水溶性的问题，利用共价固定方法解决小分子辅酶的留存问题，无细胞抽提液中内源性辅酶再生酶的利用等。除此之外，对每种辅酶再生方法与体系在工业催化中应用的优势与局限性作了评述，特别关注了辅酶再生体系反复多批次利用的问题。

关键词: 手性化合物, 辅酶再生体系, 酶耦合反应, 细胞耦合, 内源性辅酶再生酶