Hwang Bum-Yeol, Oh Jong-Min, Kim Juhan, Kim Byung-Gee
School of Chemical and Biological Engineering and Institute of Molecular Biology and Genetics, Seoul National University, Seoul, 151-742, Korea.
Biotechnol Lett. 2006 Aug;28(15):1181-5. doi: 10.1007/s10529-006-9069-8. Epub 2006 Jul 1.
A new growth-based screening method for the identification of enantioselective hydrolases, such as lipases and esterases, using pro-antibiotic substrates was devised. An enantioselective hydrolase could be identified by measuring growth rates of cells in liquid media containing (R)- or (S)-2-phenylbutyric chloramphenicol esters. This method can be applied to the screening of novel enantioselective microbes and to the high-throughput screening for the directed evolution of enantioselective hydrolytic enzymes.
设计了一种基于生长的新筛选方法,用于使用前体抗生素底物鉴定对映选择性水解酶,如脂肪酶和酯酶。通过测量含有(R)-或(S)-2-苯基丁酸氯霉素酯的液体培养基中细胞的生长速率,可以鉴定对映选择性水解酶。该方法可应用于新型对映选择性微生物的筛选以及对映选择性水解酶定向进化的高通量筛选。