Screening of Exiguobacterium acetylicum from Soil Samples Showing Enantioselective and Alkalotolerant Esterase Activity

  • Hwang Bum-Yeol (Institute for Molecular Biology and Genetics, and School of Chemical Engineering, Seoul National University) ;
  • Kim Ji-Hyun (Institute for Molecular Biology and Genetics, and School of Chemical Engineering, Seoul National University) ;
  • Kim Juhan (Institute for Molecular Biology and Genetics, and School of Chemical Engineering, Seoul National University) ;
  • Kim Byung-Gee (Institute for Molecular Biology and Genetics, and School of Chemical Engineering, Seoul National University)
  • Published : 2005.08.01

Abstract

About 3,000 bacterial colonies with esterase activities were isolated from soil samples by enrichment culture and halo-size on Luria broth-tributyrin (LT) plates. The colonies were assayed for esterase activity in microtiter plates using enantiomerically pure (R)- and (S)-2-phenylbutyric acid resorufin ester (2PB-O-res) as substrates. Two enantioselective strains (JH2 and JH13) were selected by the ratio of initial rate of hydrolysis of enantiomerically pure (R)- and (S)-2-PB-O-res. When cell pellets were used, both strains showed high apparent enantioselectivity ($E_{app}>100$) for (R)-2PB-O-res and were identified as Exiguobacterium acetylicum. The JH13 strain showed high esterase activity on p-nitrophenyl acetate (pNPA), but showed low lipase activity on p-nitrophenyl palmitate (pNPP). The esterase was located in the soluble fraction of the cell extract. The crude intracellular enzyme preparation was stable at a pH range from 6.0 to 11.0.

Keywords

References

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