Vibrio sp. GMD509의 신규 esterase의 ofloxacin ester의 광학특이성 분해 특성분석

Abstract

Vibrio sp. GMD509, a lipolytic marine bacterium isolated from the egg of sea hare. The esterase-encoding gene was cloned by screening genomic library of Vibrio sp. GMD509 and the sequence was determined. Similarity analysis revealed that the esterase gene consisted of 1017 bp (338 aa) with an estimated molecular mass of 37 kDa. The consensus sequence, G-X1-S-X2-G found in most serine esterase/lipases was also conserved, and the comparison of the deduced amino acids showed a high similarity to the enzymes of dienelactone hydrolase family. The esterase was overexpressed in E. coli and purified to homogeneity. The purified enzyme (rVlip509) was applied to the chiral resolution process of ofloxacin ester to test the potential usage as a biocatalyst. As a result, rVlip509 could hydrolyzed preferentially (S)-ofloxacin ester with eep value of 42.9 %. The novel esterase in this study might be effectively used as a novel biocatalyst for production of levofloxacin in the pharmaceutical industry.