A new high-throughput screening method for determining active and enantioselective hydrolases

作者：Wang Bo; Tang Xiaoling; Ren Gangfeng; Liu Ji; Yu Hongwei^*

来源：Biochemical Engineering Journal, 2009, 46(3): 345-349.

DOI：10.1016/j.bej.2009.06.002

• Biocatalyst preparation
• Enzyme
• Microbial
• Protein engineering
• High-throughput screening
• Fluorescein sodium salt

A new high-throughput screening method using. fluorescein sodium salt as an indicator to obtain hydrolases with high enantioselectivity is developed, which is demonstrated to be sensitive and reliable. The results determined by the method correlate well with those from GC analysis. This method can be applied to determine activity and enantioselectivity of not only lipase and esterase, but also other enzymes which catalyze hydrolysis reaction releasing proton, such as the protease or amidase. Because of the application of small amount of optically pure enantiomers, screening large libraries of enzymes is allowed at low cost and in short time.

• 出版日期2009-11-1
• 单位浙江大学