Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, China.
Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, China.
Enzyme Microb Technol. 2019 Nov;130:109362. doi: 10.1016/j.enzmictec.2019.109362. Epub 2019 Jun 17.
Transaminases (TAs, EC 2.6.1.X) are a class of promising catalysts for the synthesis of chiral amines. The addition of exogenous expensive cofactor, pyridoxal 5'-phosphate (PLP) leads to the high cost and complicated reaction composition and thus limits the industrial implementation of TAs. In this study, a novel self-sufficient biocatalyst was developed based on transaminase and PLP covalent co-immobilization with the activity recovery of 83.6% and the specific activity of 343.0 U/g catalyst. The self-sufficient BgTA biocatalyst was employed in the continuous biosynthesis of (R)-sitagliptin in a recirculating packed bed reactor (RPBR) for 700 batches reaction without extra addition of PLP. The yield and e.e. of (R)-sitagliptin for each batch were above 90% and 99% respectively. The space-time yield was 40.0 g/(L·h) which was higher than ever reported. This is the first report about the covalent co-immobilization of TA and PLP with high activity and stability, and our work also contributed to the economic flexibility of chiral amine pharmaceutical intermediates biosynthesis.
转氨酶（TAs，EC 2.6.1.X）是一类很有前途的手性胺合成催化剂。外源性昂贵辅因子吡哆醛 5'-磷酸（PLP）的添加导致成本高、反应成分复杂，从而限制了 TAs 的工业实施。在本研究中，基于转氨酶和 PLP 的共价共固定化开发了一种新型自给型生物催化剂，其活性回收率为 83.6%，比酶活为 343.0 U/g 催化剂。自给型 BgTA 生物催化剂在循环填充床反应器（RPBR）中连续合成（R）-西他列汀，在没有额外添加 PLP 的情况下进行了 700 批反应。每个批次的（R）-西他列汀的产率和对映体过量值（ee 值）均高于 90%和 99%。时空产率为 40.0 g/（L·h），高于以往报道的任何结果。这是首次报道 TA 和 PLP 的共价共固定化具有高活性和稳定性，我们的工作也为手性胺药物中间体生物合成的经济灵活性做出了贡献。
