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A unique polypeptide from the C-terminus of the exocellular esterase of Acinetobacter venetianus RAG-1 modulates the emulsifying activity of the polymeric bioemulsifier apoemulsan

Applied Microbiology and Biotechnology volume 71, pages 177–183 (2006)Cite this article

Abstract

An exocellular esterase from the oil-degrading Acinetobacter venetianus RAG-1 was previously shown to enhance the emulsification and emulsion stabilization properties of the amphipathic, aminopolysaccharide bioemulsifier, emulsan [Bach H, Berdichevsky Y, Gutnick D (2003) An exocellular protein from the oil-degrading microbe Acinetobacter venetianus RAG-1 enhances the emulsifying activity of the polymeric bioemulsifier emulsan. Appl Environ Microbiol 69:2608–15]. This enhancement was specific for the RAG-1 esterase and was independent of catalytic activity. In this report, fragments from both the N′- and C′-termini were cloned as fusions to the C-terminus of the maltose-binding protein (MBP) and were tested for enhancement activity in the presence of the deproteinated form of emulsan, apoemulsan. The activity could be localized to the C-terminal third of the protein which exhibited the same activity as the intact enzyme. MBP itself was completely inactive and could be cleaved from the fusion without affecting the subsequent emulsification. However, the enhancement completely depended on the presence of a unique C-terminal 20 amino acid peptide not found in any other protein in the databases. In addition, progressive removal of amino acids from the N-terminus of the active MBP polypeptide resulted in a concomitant loss of activity, indicating that enhancement is also proportional to the size of the peptide fragment. The middle third and the C-terminal third of the enzyme each contained a copy of the conserved Cardin–Weintraub consensus sequence for protein binding to heparin. These sequences were not detected in homologous esterases from a closely related strain, Acinetobacter calcoaceticus BD413.

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References

  • Alon RN, Gutnick DL (1993) Esterase from the oil-degrading Acinetobacter lwoffii RAG-1: sequence analysis and over-expression in Escherichia coli. FEMS Microbiol Lett 112:275–280

    CAS  Article  Google Scholar 

  • Alon RN, Mirny L, Sussman JL, Gutnick DL (1995) Detection of α/β-hydrolase fold in the cell surface esterases of Acinetobacter species using an analysis of 3D profiles. FEBS Lett 371:231–235

    CAS  Article  Google Scholar 

  • Aricescu AR, McKinnell IW, Halfter W, Stoker AW (2002) Heparan sulfate proteoglycans are ligands for receptor protein tyrosine phosphatase sigma. Mol Cell Biol 22:1881–1892

    CAS  Article  Google Scholar 

  • Armstrong CG, Doherty MJ, Cohen PT (1998) Identification of the separate domains in the hepatic glycogen-targeting subunit of protein phosphatase 1 that interact with phosphorylase a, glycogen and protein phosphatase 1. Biochem J 336(Pt 3):699–704

    CAS  Article  Google Scholar 

  • Bach H, Gutnick DL (2004) Potential applications of bioemulsifiers in the oil industry. In: Vazquez-Duhalt R, Quintero-Ramírez R (eds) Petroleum biotechnology, developments and perspectives. Elsevier, The Netherlands, pp. 233–281

    Chapter  Google Scholar 

  • Bach H, Berdichevsky Y, Gutnick D (2003) An exocellular protein from the oil-degrading microbe Acinetobacter venetianus RAG-1 enhances the emulsifying activity of the polymeric bioemulsifier emulsan. Appl Environ Microbiol 69:2608–2615

    CAS  Article  Google Scholar 

  • Belsky I, Gutnick DL, Rosenberg E (1979) Emulsifier of Arthrobacter RAG-1: determination of emulsifier-bound fatty acids. FEBS Lett 1979;101:175–178

    CAS  Article  Google Scholar 

  • Cardin AD, Weintraub HJ (1989) Molecular modeling of protein–glycosaminoglycan interactions. Arteriosclerosis 9:21–32

    CAS  Article  Google Scholar 

  • Chen L, Wu Y, Lee V, Kiani C, Adams ME, Yao Y, Yang BB (2002) The folded modules of aggrecan G3 domain exert two separable functions in glycosaminoglycan modification and product secretion. J Biol Chem 277:2657–2665

    CAS  Article  Google Scholar 

  • Coutinho JB, Gilkes NR, Warren RA, Kilburn DG, Miller Jr RC (1992) The binding of Cellulomonas fimi endoglucanase C (CenC) to cellulose and Sephadex is mediated by the N-terminal repeats. Mol Microbiol 6:1243–1252

    CAS  Article  Google Scholar 

  • Desai JD, Banat IM (1997) Microbial production of surfactants and their commercial potential. Microbiol Mol Biol Rev 61:47–64

    CAS  Article  Google Scholar 

  • Din N et al (1994) The cellulose-binding domain of endoglucanase A (CenA) from Cellulomonas fimi: evidence for the involvement of tryptophan residues in binding. Mol Microbiol 11:747–755

    CAS  Article  Google Scholar 

  • Ferreira LM, Wood TM, Williamson G, Faulds C, Hazlewood GP, Black GW, Gilbert HJ (1993) A modular esterase from Pseudomonas fluorescens subsp. cellulosa contains a non-catalytic cellulose-binding domain. Biochem J 294(Pt 2):349–355

    CAS  Article  Google Scholar 

  • Fromm JR, Hileman RE, Caldwell EE, Weiler JM, Linhardt RJ (1997) Pattern and spacing of basic amino acids in heparin binding sites. Arch Biochem Biophys 343:92–100

    CAS  Article  Google Scholar 

  • Gutierrez R, Cederlund E, Hjelmqvist L, Peirano A, Herrera F, Ghosh D, Duax W, Jornvall H, Eyzaguirre J (1998) Acetyl xylan esterase II from Penicillium purpurogenum is similar to an esterase from Trichoderma reesei but lacks a cellulose binding domain. FEBS Lett 423:35–38

    CAS  Article  Google Scholar 

  • Gutnick DL, Minas W (1987) Perspectives on microbial surfactants. Biochem Soc Trans 15(Suppl):22S

    CAS  PubMed  Google Scholar 

  • Hata A, Ridinger DN, Sutherland S, Emi M, Shuhua Z, Myers RL, Ren K, Cheng T, Inoue I, Wilson DE (1993) Binding of lipoprotein lipase to heparin. Identification of five critical residues in two distinct segments of the amino-terminal domain. J Biol Chem 268:8447–8457

    CAS  PubMed  Google Scholar 

  • Hileman RE, Fromm JR, Weiler JM, Linhardt RJ (1998) Glycosaminoglycan–protein interactions: definition of consensus sites in glycosaminoglycan binding proteins. Bioessays 20:156–167

    CAS  Article  Google Scholar 

  • Kaplan N, Zosim Z, Rosenberg E (1987) Reconstitution of emulsifying activity of Acinetobacter calcoaceticus BD4 emulsan by using pure polysaccharide and protein. Appl Environ Microbiol 53:440–446

    CAS  Article  Google Scholar 

  • Kassam G, Manro A, Braat CE, Louie P, Fitzpatrick SL, Waisman DM (1997) Characterization of the heparin binding properties of annexin II tetramer. J Biol Chem 272:15093–15100

    CAS  Article  Google Scholar 

  • Kok RG, Christoffels VM, Vosman B, Hellingwerf KJ (1993) Growth-phase-dependent expression of the lipolytic system of Acinetobacter calcoaceticus BD413: cloning of a gene encoding one of the esterases. J Gen Microbiol 139:2329–2342

    CAS  Article  Google Scholar 

  • Lookene A, Nielsen MS, Gliemann J, Olivecrona G (2000) Contribution of the carboxy-terminal domain of lipoprotein lipase to interaction with heparin and lipoproteins. Biochem Biophys Res Commun 271:15–21

    CAS  Article  Google Scholar 

  • Reddy PG, Allon R, Mevarech M, Mendelovitz S, Sato Y, Gutnick DL (1989) Cloning and expression in Escherichia coli of an esterase-coding gene from the oil-degrading bacterium Acinetobacter calcoaceticus RAG-1. Gene 76:145–152

    CAS  Article  Google Scholar 

  • Reisfeld A, Rosenberg E, Gutnick DL (s1972) Microbial degradation of crude oil: factors affecting the dispersion in sea water by mixed and pure cultures. Appl Microbiol 24:363–368

    CAS  Article  Google Scholar 

  • Rosenberg E, Zuckerberg A, Rubinovitz C, Gutnick DL (1972) Emulsifier of Arthrobacter RAG-1: isolation and emulsifying properties. Appl Environ Microbiol 37:402–408

    CAS  Article  Google Scholar 

  • Sambrook JF, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual. 2nd ed. Cold Spring Harbor Laboratory Press, New York

    Google Scholar 

  • Shabtai Y, Gutnick DL. Exocellular esterase and emulsan release from the cell surface of Acinetobacter calcoaceticus. J Bacteriol 1985;161:1176–1181

    CAS  Article  Google Scholar 

  • Shabtai Y, Pines O, Gutnick DL. Emulsan: a case study of microbial capsules as industrial products. Dev Ind Microbiol 1986;26:291–307

    Google Scholar 

  • Toren A, Navon V, Ron EZ, Rosenberg E (2001) Emulsifying activities of purified alasan proteins from Acinetobacter radioresistens KA53. Appl Environ Microbiol 67:1102–1106

    CAS  Article  Google Scholar 

  • Vincent P, Shareck F, Dupont C, Morosoli R, Kluepfel D (1997) New alpha-L-arabinofuranosidase produced by Streptomyces lividans: cloning and DNA sequence of the abfB gene and characterization of the enzyme. Biochem J 322(Pt 3):845–852

    CAS  Article  Google Scholar 

  • Wang J, Stuckey JA, Wishart MJ, Dixon JE (2002) A unique carbohydrate binding domain targets the lafora disease phosphatase to glycogen. J Biol Chem 277:2377–2380

    CAS  Article  Google Scholar 

  • Watanabe T, Kasahara N, Aida K, Tanaka H (1992) Three N-terminal domains of beta-1,3-glucanase A1 are involved in binding to insoluble beta-1,3-glucan. J Bacteriol 174:186–190

    CAS  Article  Google Scholar 

  • Wu J, Liu J, Thompson I, Oliver CJ, Shenolikar S, Brautigan DL (1998) A conserved domain for glycogen binding in protein phosphatase-1 targeting subunits. FEBS Lett 439:185–191

    CAS  Article  Google Scholar 

  • Yamamoto M, Ezure T, Watanabe T, Tanaka H, Aono R (1998) C-terminal domain of beta-1,3-glucanase H in Bacillus circulans IAM1165 has a role in binding to insoluble beta-1,3-glucan. FEBS Lett 433:41–43

    CAS  Article  Google Scholar 

  • Zosim Z, Gutnick D, Rosenberg E (1982) Properties of hydrocarbon-in-water emulsions stabilized by Acinetobacter RAG-1 emulsan. Biotechnol Bioeng 24:281–292

    CAS  Article  Google Scholar 

  • Zuckerberg A, Diver A, Peeri Z, Gutnick DL, Rosenberg E (1979) Emulsifier of Arthrobacter RAG-1: chemical and physical properties. Appl Environ Microbiol 37:414–420

    CAS  Article  Google Scholar 

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Acknowledgements

This work was supported in part by a grant from the CDR program of the US AID. We thank Rina Avigad for excellent technical assistance and David Nakar for constructive suggestions and advice throughout the course of this research. This work is in partial satisfaction of the requirements for the Ph.D. degree in Microbiology awarded to H.B. at Tel-Aviv University.

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  1. Division of Infectious Diseases, Vancouver General Hospital, 2733 Heather St.,, Vancouver, BC, V5Z 3J5, Canada

    Horacio Bach

  2. Dept. of Molecular Microbiology and Biotechnology, Faculty of Life Science, Tel Aviv University, Ramat Aviv, 69978, Israel

    David L. Gutnick

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  1. Horacio Bach
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Correspondence to David L. Gutnick.

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Bach, H., Gutnick, D.L. A unique polypeptide from the C-terminus of the exocellular esterase of Acinetobacter venetianus RAG-1 modulates the emulsifying activity of the polymeric bioemulsifier apoemulsan. Appl Microbiol Biotechnol 71, 177–183 (2006). https://doi.org/10.1007/s00253-005-0161-0

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  • DOI: https://doi.org/10.1007/s00253-005-0161-0

Keywords

  • Emulsification
  • Amino Acid Peptide
  • Acinetobacter Calcoaceticus
  • Column Buffer
  • Recombinant Esterase