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Biocatalysed synthesis of chiral amines: continuous colorimetric assays for mining amine-transaminases

机译：Biocatalysed合成手性胺:连续采矿的比色测定amine-transaminases

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In the course of our research aimed at the design of new biocatalytic processes for the enantioselective synthesis of chiral amines, we have developed new continuous assays for the screening of amine-transaminase collections. These assays are based on the use of hypotaurine as an irreversible amine donor. This beta-aminosulfinic acid is converted upon transamination into 2-oxoethylsulfinic acid, which instantaneously decomposes into acetaldehyde and sulfite ions that can be easily detected by spectrophotometry using Ellman's reagent. Two complementary assays were developed based on this titration method. Firstly, a direct assay allowed detection of various transaminases able to use hypotaurine as an amino donor. In a second coupled assay, l-alanine is used as a generic donor substrate of amine-transaminases and is regenerated using an auxiliary hypotaurine-transaminase. The powerful and complementary nature of both assays was demonstrated through the screening of a collection of 549 amine-transaminases from biodiversity, thus allowing the discovery of a variety of valuable new biocatalysts for use in synthetic processes.
机译：在我们的研究旨在设计新的biocatalytic流程拆分的合成手性胺,我们开发了新的连续化验吗筛选amine-transaminase集合。这些分析都是基于hypotaurine的使用作为一个不可逆转的胺捐献者。beta-aminosulfinic酸转化转氨作用到2-oxoethylsulfinic酸,瞬间分解成乙醛和亚硫酸盐离子很容易检测到使用Ellman的分光光度法试剂基于这个滴定方法。分析允许各种氨基转移酶的检测能够使用hypotaurine作为氨基供体。第二耦合分析,丙氨酸作为通用amine-transaminases供体基质使用一个辅助和再生hypotaurine-transaminase。互补性的化验证明了通过筛选549 amine-transaminases的集合生物多样性,从而允许的发现各种有价值的新生物催化剂在使用合成过程。

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