Computational and experimental identification of an exceptionally efficient ethyl ester synthetase with broad...

Transesterification plays a crucial role in the synthesis of diverse esters in organic synthesis but is barely reported in biocatalysis. Here, we computationally identify the salicylic acid-binding protease 2 (SABP2) as an efficient ethyl ester bond synthetase by QM/MM MD and free energy simulations and present the practical and effective utilization of SABP2 as an eco-friendly biocatalyst for transesterification reactions by a series of experiments. Our findings demonstrate that SABP2 efficiently catalyzes the transesterification reaction between the carboxyl acid group of promiscuous aromatic substrates and ethanol to produce the corresponding ethyl esters. Notably, while SABP2 exhibits its native capability to catalyze the hydrolysis of the methyl salicylate (MeSA), the transesterification rate (producing ethyl salicylate, EtSA) is about 3500 times higher than the hydrolysis rate. Additionally, a range of aromatic methyl esters are employed in the transesterification process, resulting in high yields (up to 98.9 %) of the corresponding ethyl esters. These results indicate a broad substrate scope for SABP2-catalyzed transesterification reactions, demonstrating its potential as a valuable biocatalyst for ester synthesis in industry.