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Production and Purification of a Solvent-Resistant Esterase from Bacillus licheniformis S-86

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Abstract

New thermophilic and organic-solvent-tolerant Bacillus licheniformis S-86 strain is able to produce two active and solvent-stable esterases. Production of type I and II esterases was substantially enhanced when oils and surfactants were supplied as carbon sources. Grape oil (0.1% v/v) and Tween 20 to 60 (0.1% v/v) had enhanced enzyme production between 1.6- and 2.2-folds. Type II esterase was purified to homogeneity in a five-step procedure. This esterase was purified 76.7-fold with a specific activity of 135 U mg−1. Molecular mass of the enzyme was estimated to be 38.4 kDa by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Type II esterase was active mostly on esters with short acyl chains, which allowed to classify the enzyme as a carboxylesterase with a K m of 80.2 mmol l−1 and a V max of 256.4 μmol min−1 mg−1 for p-nitrophenyl acetate. Also, B. licheniformis S-86 type II esterase displayed activity in presence of water-miscible organic solvents at 50% concentration and stability after 1-h incubation.

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Acknowledgments

The authors would like to thank E. Rodriguez for excellent technical assistance. The financial support from Pew Charitable Trust (USA), CONICET (PIP 6203/06, Argentina) and ANPCyT (BID N° 1728/ OC-AR) is gratefully acknowledged.

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Authors and Affiliations

  1. Planta Piloto de Procesos Industriales Microbiológicos (PROIMI), Av. Belgrano y Pasaje Caseros, T4001, MVB, Tucumán, Argentina

    Sebastián Torres & Mario D. Baigorí

  2. National Institute for Interdisciplinary Science and Technology, Trivandrum, 695 019, India

    Ashok Pandey

  3. CINDEFI (Institute of Applied Biotechnology) and INIFTA (CONICET), School of Sciences, La Plata University, Calle 50 y 115, CP 1900, La Plata, Argentina

    Guillermo R. Castro

  4. Department of Biomedical Engineering, Tufts University, 4 Colby Street, Medford, MA, 02155, USA

    Guillermo R. Castro

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  1. Sebastián Torres
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  2. Mario D. Baigorí
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Correspondence to Guillermo R. Castro.

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Torres, S., Baigorí, M.D., Pandey, A. et al. Production and Purification of a Solvent-Resistant Esterase from Bacillus licheniformis S-86. Appl Biochem Biotechnol 151, 221–232 (2008). https://doi.org/10.1007/s12010-008-8181-8

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