Amine transaminases (ATAs) are powerful enzymes for the synthesis of chiral amines. Although the request for amines with more than one chiral center is increasing, their synthesis is still challenging. Here we show a casacde reaction combining an enoate reductase (ERED) and an amine transaminase (ATA-VibFlu), which allows access to optically pure (1R,3R)-1-amino-3-methylcyclohexane. Because all known wildtype EREDs show a (S)-selectivity for 3-methylcyclohex-2-enone and the ATA-VibFlu only showed a modest enantioselectivity, different variants of EREDs and ATAs were investigated and suitable mutant enzymes were identified. In whole cell biocatalyses using the ERED YqjM Cys26Asp/Ile69Thr and the ATA-VibFlu Leu56Ile (1R,3R)-1-amino-3-methylcyclohexane was obtained at high optical purity (97% de).

中文翻译：

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酶联反应合成（1R，3R）-1-氨基-3-甲基环己烷

胺转氨酶（ATAs）是用于合成手性胺的强大酶。尽管对具有多个手性中心的胺的需求在增加，但其合成仍具有挑战性。在这里，我们显示了一个结合了烯醇酸还原酶（ERED）和胺转氨酶（ATA-VibFlu）的casacde反应，该反应可以访问光学纯的（1R，3R）-1-氨基-3-甲基环己烷。由于所有已知的野生型ERED对3-甲基环己-2-烯酮均显示（S）选择性，而ATA-VibFlu仅显示适度的对映选择性，因此对ERED和ATA的不同变体进行了研究，并确定了合适的突变酶。使用ERED YqjM Cys26Asp / Ile69Thr和ATA-VibFlu Leu56Ile（1R，3R）-1-氨基-3-甲基环己烷以高光学纯度（97％de）获得全细胞生物催化。