Transaminase Engineering and Process Development for a Whole-Cell Neat Organic Process to Produce (R)‑α-Phenylethylamine

The production of (R)-α-phenylethylamine ((R)-α-PEA) from acetophenone is a classic reaction for the characterization of transaminases. However, developing a commercially viable transaminase process to manufacture (R)-α-PEA usually suffers from two drawbacks. One is related to the biocatalyst itself...

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Published in:Organic process research & development 2022-07, Vol.26 (7), p.2004-2012
Main Authors: Cai, Baoqin, Wang, Jiyong, Hu, Hu, Liu, Sitong, Zhang, Chengxiao, Zhu, Ying, Bocola, Marco, Sun, Lei, Ji, Yaoyao, Zhou, Ameng, He, Kuifang, Peng, Qinli, Luo, Xiao, Hong, Ruimei, Wang, Juanjuan, Shang, Chuanyang, Wang, Zikun, Yang, Zhuhong, Bong, Yong Koy, Daussmann, Thomas, Chen, Haibin
Format: Article
Language:English
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Summary:The production of (R)-α-phenylethylamine ((R)-α-PEA) from acetophenone is a classic reaction for the characterization of transaminases. However, developing a commercially viable transaminase process to manufacture (R)-α-PEA usually suffers from two drawbacks. One is related to the biocatalyst itself, since transaminases are easily inhibited by (R)-α-PEA at low concentrations. The other drawback is a common low space-time yield of typical transaminase processes, because the reaction equilibrium greatly favors the formation of acetophenone over (R)-α-PEA. In this study, an (R)-selective amine transaminase (TA) from Aspergillus fumigatus Af293 was engineered by a directed evolution for an efficient process to produce (R)-α-PEA. The evolved variant showed an over 3000-fold increase in activity and a tolerance with 2.0 M isopropylamine as well as the complete absence of inhibition by (R)-α-PEA. At the same time, using this evolved TA variant, a continuous neat organic process using whole-cells was developed where the biocatalyst and remaining acetophenone can be efficiently separated from (R)-α-PEA and reused repetitively. This not only decreases the overall cost and waste generation but also achieves a very high space-time yield of up to 168 g L–1 d–1 of (R)-α-PEA in an industrial pilot scale setup.
ISSN:1083-6160
1520-586X
DOI:10.1021/acs.oprd.1c00409