Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, People's Republic of China; Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou 310014, People's Republic of China.
Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, People's Republic of China; Engineering Research Center of Bioconversion and Biopurification of Ministry of Education, Zhejiang University of Technology, Hangzhou 310014, People's Republic of China.
J Biotechnol. 2019 Apr 10;295:49-54. doi: 10.1016/j.jbiotec.2019.02.008. Epub 2019 Mar 7.
(R)-3-amino-1-butanol is a key intermediate of Dolutegravir for the treatment of HIV/AIDS and its green and efficient biosynthesis has attracted a great deal of attention. Transaminases are currently used as promising biocatalyst for the synthesis of chiral amines. However, many transaminases have (S)-specificity and (R)-selective transaminases were less exploited and studied, making the production of (R)-amines remain challenging. In this study, a novel transaminase from Actinobacteria sp. (As-TA) was obtained and applied for the biosynthesis of (R)-3-amino-1-butanol by transferring the amino group from isopropylamine to 4-hydroxy-2-butanone. After optimization of the reaction condition and using a substrate fed-batch strategy, the conversion of 100, 200, 300, 400 and 500 mM 4-hydroxy-2-butanone reached 100%, 94.9%, 86.1%, 76.1% and 70.9%, respectively. (R)-3-amino-1-butanol with a maximum yield of 29.6 g/L and 99.9% e.e. value was obtained. This was the first time demonstrating the successful biosynthesis of (R)-3-amino-1-butanol with transaminase as biocatalyst and the obtained As-TA enriched the enzyme pool of transaminase with (R)-specificity.
（R）-3-氨基-1-丁醇是治疗 HIV/AIDS 的多替拉韦的关键中间体，其绿色高效的生物合成引起了广泛关注。转氨酶目前被用作手性胺合成的有前途的生物催化剂。然而，许多转氨酶具有（S）-特异性，而（R）-选择性转氨酶的研究和利用较少，使得（R）-胺的生产仍然具有挑战性。在这项研究中，从放线菌属（As-TA）获得了一种新型转氨酶，并通过将氨基从异丙胺转移到 4-羟基-2-丁酮来应用于（R）-3-氨基-1-丁醇的生物合成。在优化反应条件并采用底物补料分批策略后，分别达到 100、200、300、400 和 500 mM 4-羟基-2-丁酮的转化率为 100%、94.9%、86.1%、76.1%和 70.9%。（R）-3-氨基-1-丁醇的最大产量为 29.6 g/L，ee 值为 99.9%。这是首次成功地使用转氨酶作为生物催化剂合成（R）-3-氨基-1-丁醇，获得的 As-TA 丰富了具有（R）-特异性的转氨酶酶库。
