Department of Systems Biotechnology, Konkuk University, Seoul, Gwangjin-gu, South Korea.
CKD Bio Research Institute, Ansan-si, Gyeonggi-do, South Korea.
Biotechnol Bioeng. 2021 Aug;118(8):3263-3268. doi: 10.1002/bit.27819. Epub 2021 May 21.
Here, we report a bienzymatic cascade to produce β-amino acids as an intermediate for the synthesis of the leading oral antidiabetic drug, sitagliptin. A whole-cell biotransformation using recombinant Escherichia coli coexpressing a esterase and transaminase were developed, wherein the desired expression level of each enzyme was achieved by promotor engineering. The small-scale reactions (30 ml) performed under optimized conditions at varying amounts of substrate (100-300 mM) resulted in excellent conversions of 82%-95% for the desired product. Finally, a kilogram-scale enzymatic reaction (250 mM substrate, 220 L) was carried out to produce β-amino acid (229 mM). Sitagliptin phosphate was chemically synthesized from β-amino acids with 82% yield and > 99% purity.
在这里，我们报告了一种双酶级联反应，用于生产β-氨基酸作为合成领先的口服抗糖尿病药物西他列汀的中间体。使用共表达酯酶和转氨酶的重组大肠杆菌进行全细胞生物转化，其中通过启动子工程实现了每种酶的所需表达水平。在优化条件下，在不同底物量（100-300 mM）下进行小规模反应（30 ml），导致所需产物的转化率达到 82%-95%。最后，进行了公斤级酶反应（250 mM 底物，220 L）以生产β-氨基酸（229 mM）。西他列汀磷酸盐通过β-氨基酸的化学合成得到，收率为 82%，纯度>99%。
