Abstract
An esterase PE10 (279 aa) from Pelagibacterium halotolerans B2T was cloned and overexpressed in Escherichia coli Rosetta in a soluble form. The deduced protein was 29.91 kDa and the phylogenetic analysis of the deduced amino acids sequence showed it represented a new family of lipolytic enzymes. The recombinant protein was purified by Ni–NTA affinity chromatography column and the characterization showed its optimal temperature and pH were 45 °C and pH 7.5, respectively. Substrate specificity study showed PE10 preferred short chain p-nitrophenyl esters and exhibited maximum activity toward p-nitrophenyl acetate. In addition, PE10 was a halotolerant esterase as it was still active under 4 M NaCl. Three-dimensional modeling of PE10 suggested that the high negative electrostatic potential on the surface may relevant to its tolerance to high salt environment. With this halotolerance property, PE10 could be a candidate for industrial use.
Similar content being viewed by others
References
Akoh CC, Lee GC, Liaw YC, Huang TH, Shaw JF (2004) GDSL family of serine esterases/lipases. Prog Lipid Res 43:534–552
Arpigny JL, Jaeger KE (1999) Bacteria lipolytic enzymes: classification and properties. Biochem J 343:177–183
Bornscheuer UT (2001) Microbial carboxyl esterase: classification, properties and application in biocatalysis. FEMS Microbiol Rev 26:73–81
Bradford MM (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72:248–254
Cao Y, Liao L, Xu XW, Oren A, Wang C, Zhu XF, Wu M (2008) Characterization of alcohol dehydrogenase from the haloalkaliphilic archaeon Natronomonas pharaonis. Extremophiles 12:471–476
Danson MJ, Hough DW (1997) The structural basis of protein halophilicity. Comp Biochem Physiol 117:307–312
De Simone G, Mandrich L, Menchise V, Giordano V, Febbraio F, Rossi M, Pedone C, Manco G (2004) A substrate-induced switch in the reaction mechanism of a thermophilic esterase. J Biol Chem 279:6815–6823
Hess M, Katzer M, Antranikian G (2008) Extremely thermostable esterases from the thermoacidophilic euryarchaeon Picrophilus torridus. Extremophiles 12:351–364
Huo YY, Cheng H, Han XF, Jiang XW, Sun C, Zhang XQ, Zhu XF, Liu YF, Li PF, Ni PX, Wu M (2012) Complete genome sequence of Pelagibacterium halotolerans B2T. J Bacteriol 194:197–198
Jeanmougin F, Thompson JD, Gouy M, Higgins DG, Gibson TJ (1998) Multiple sequence alignment with Clustal X. Trends Biochem Sci 23:403–405
Jiao N, Zhang R, Zheng Qiang (2010) Coexistence of two different photosynthetic operons in Citromicrobium bathyomarinum JL354 as revealed by whole-genome sequencing. J Bacteriol 192:1169–1170
Khudary RA, Venkatachalam R, Katzer M, Elleuche S, Antranikian G (2010) A cold-adapted esterase of a novel marine isolate, Pseudoalteromonas arctica: gene cloning, enzyme purification and characterization. Extremophiles 14:273–285
Kim J, Dordick JS (1997) Unusual salt and solvent dependence of a protease from an extreme halophile. Biotechnol Bioeng 55:471–479
Kim JY, Choi GS, Kim YJ, Ryu YW, Kim GJ (2002) A new isolate Bacillus stearothermophilus JY144 expressing a novel esterase with high enantioselectivity to (R)-ketoprofen ethyl ester: strain selection and gene cloning. J Mol Catal B 18:133–145
Kim JY, Choi GS, Kim YJ, Lee JB, Jo DH, Ryu YW (2003) Screening, production and properties of a stereospecific esterase from Pseudomonas sp. S34 with high selectivity to (S)-ketoprofen ethyl ester. J Mol Catal B 22:29–35
Kushner DJ (1988) What is the “true” internal environment of halophilic and other bacteria? Can J Microbiol 34:482–486
Kwoun KH, Jung YJ, Choi WC, Ryu HS, Oh TK, Lee JK (2004) Sequence-based approach to finding functional lipases from microbial genome databases. FEMS Microbiol Lett 235:349–355
Lauro FM, McDougald D, Thomas T, Williams TJ, Egan S, Rice S, DeMaere MZ, Ting L, Ertan H, Johnson J, Ferriera S, Lapidus A, Anderson I, Kyrpides N, Munk AC, Detter C, Han CS, Brown MV, Robb FT, Kjelleberg S, Cavicchioli R (2009) The genomic basis of trophic strategy in marine bacteria. Proc Natl Acad Sci 106:15527–15533
Miller TR, Delcher AL, Salzberg SL, Saunders E, Detter JC, Halden RU (2010) Genome sequence of the dioxin-mineralizing bacterium Sphingomonas wittichii RW1. J Bacteriol 192:6101–6102
Müller-Santos M, Souza EMD, Pedrosa FO, Mitchell DA, Longhi S, Carrière F, Canaan S, Krieger N (2009) First evidence for the salt-dependent folding and activity of an esterase from the halophilic archaea Haloarcula marismortui. BBA Mol Cell Bio L 1791:719–729
Nierman WC, Feldblyum TV, Laub MT, Paulsen IT, Nelson KE, Eisen J, Heidelberg JF, Alley MRK, Ohta N, Maddocki JR, Potocka I, Nelson WC, Newton A, Stephens C, Phadkei ND, Ely B, DeBoy RT, Dodson RJ, Durkin AS, Gwinn ML, Haft DH, Kolonay JF, Smit J, Craven MB, Khouri H, Shetty J, Berry K, Utterback T, Tran K, Wolf A, Vamathevan J, Ermolaeva M, White O, Salzberg SL, Venter JC, Shapiro L, Fraser CM (2001) Complete genome sequence of Caulobacter crescentus. Proc Natl Acad Sci 98:4136–4141
Oh HM, Giovannoni SJ, Ferriera S, Johnson J, Chol JC (2009) Complete genome sequence of Erythrobacter litoralis HTCC2594. J Bacteriol 191:2419–2420
Park HJ, Jeon JH, Kang SG, Lee JH, Lee SA, Kim HK (2007) Functional expression and refolding of new alkaline esterase, EM2L8 from deep-sea sediment metagenome. Protein Expres Purif 52:340–347
Pascale DD, Cusano AM, Autore F, Parrilli E, Prisco GD, Marino G, Tutino ML (2008) The cold-active Lip1 lipase from the Antarctic bacterium Pseudoalteromonas haloplanktis TAC125 is a member of a new bacterial lipolytic enzyme family. Extremophiles 12:311–323
Rao L, Zhao X, Pan F, Li Y, Xue Y, Ma Y, Lu JR (2009) Solution behavior and activity of a halophilic esterase under high salt concentration. PLoS ONE 4:e6980
Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425
Sellek GA, Chaudhuri JB (1999) Biocatalysis in organic media using enzymes from extremophiles. Enzyme Microb Tech 25:471–482
Silby MW, Cerdeño-Tárraga AM, Vernikos GS, Giddens SR, Jackson RW, Preston GM, Zhang XX, Moon CD, Gehrig SM, Godfrey SAC, Knight CG, Malone JG, Robinson Z, Spiers AJ, Harris S, Challis GL, Yaxley AM, Harris D, Seeger K, Murphy L, Rutter S, Squares R, Quail MA, Saunders E, Mavromatis K, Brettin TS, Bentley SD, Hothersall J, Stephens E, Thomas CM, Parkhill J, Levy SB, Rainey PB, Thomson NR (2009) Genomic and genetic analyses of diversity and plant interactions of Pseudomonas fluorescens. Genome Biol 10:R51
Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599
Taylor SJC, Brown RC, Keene PA (1999) Novel screening methods—the key to cloning commercially successful biocatalysts. Bioorg Med Chem 7:2163–2168
Van den Burg B (2003) Extremophiles as a source for novel enzymes. Curr Opin Microbiol 6:213–218
Wahler D, Reymond JL (2001) Novel methods for biocatalyst screening. Curr Opin Chem Biol 5:152–158
Xu XW, Huo YY, Wang CS, Oren A, Cui HL, Vedler E, Wu M (2011) Pelagibacterium halotolerans gen. nov., sp. nov. and Pelagibacterium luteolum sp. nov., novel members of the family Hyphomicrobiaceae. Int J Syst Evol Microbiol 61:1817–1822
Acknowledgments
This work was supported by the Open Research Program of the Key Laboratory of Marine Ecosystem and Biogeochemistry (LMEB) of the State Oceanic Administration (SOA) (grant LMEB201101).
Author information
Authors and Affiliations
Corresponding author
Additional information
Communicated by A. Driessen.
Rights and permissions
About this article
Cite this article
Jiang, X., Huo, Y., Cheng, H. et al. Cloning, expression and characterization of a halotolerant esterase from a marine bacterium Pelagibacterium halotolerans B2T . Extremophiles 16, 427–435 (2012). https://doi.org/10.1007/s00792-012-0442-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00792-012-0442-3