Skip to main content
SpringerLink
Log in

The DFPase from Loligo vulgaris in sugar surfactant-based bicontinuous microemulsions: structure, dynamics, and enzyme activity

  • Original Paper
  • Published:
European Biophysics Journal Aims and scope Submit manuscript

Abstract

The enzyme diisopropyl fluorophosphatase (DFPase) from the squid Loligo vulgaris is of great interest because of its ability to catalyze the hydrolysis of highly toxic organophosphates. In this work, the enzyme structure in solution (native state) was studied by use of different scattering methods. The results are compared with those from hydrodynamic model calculations based on the DFPase crystal structure. Bicontinuous microemulsions made of sugar surfactants are discussed as host systems for the DFPase. The microemulsion remains stable in the presence of the enzyme, which is shown by means of scattering experiments. Moreover, activity assays reveal that the DFPase still has high activity in this complex reaction medium. To complement the scattering experiments cryo-SEM was also employed to study the microemulsion structure.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11
Fig. 12
Fig. 13
Fig. 14

Similar content being viewed by others

References

  • Arriaga LR, Lopez-Montero I, Monroy F, Gil GO, Farago B, Hellweg T (2009a) Stiffening effect of cholesterol on disordered lipid phases: a combined NSE + DLS analysis of the bending elasticity of large unilamellar vesicles based on POPC. Bipohys J 96:3629–3637

    Article  CAS  Google Scholar 

  • Arriaga LR, Lopez-Montero I, Monroy F, Gil GO, Farago B, Hellweg T (2009b) Fluctuation dynamics of rigid vesicles: frustration of regular bulk dissipation into a sub-diffusive relaxation. Phys Rev E 80:031908

    Article  Google Scholar 

  • Arriaga LR, Rodriguez-Garcia R, Lopez-Montero I, Farago B, Hellweg T, Monroy F (2010) Dissipative curvature fluctuations in bilayer vesicles: coexistence of pure-bending and hybrid curvature-compression modes. Euro Phys J E 31:105–115

    Article  CAS  Google Scholar 

  • Aveyard R, Binks BP, Fletcher PDI, Buscall R, Davies S (1998) Surface and colloid chemistry of systems containing pure sugar surfactants. Langmuir 14:4699–4709

    Article  CAS  Google Scholar 

  • Baier J, Koetz J, Kosmella S, Tiersch B, Rehage H (2007) Polyelectrolyte-modified inverse microemulsions and their use as templates for the formation of magnetite nanoparticles. J Phys Chem B 111(29):8612–8618

    Article  PubMed  CAS  Google Scholar 

  • Bargeron CB (1974) Measurement of continuous distribution of spherical particles by intensity correlation spectroscopy: analysis by cumulants. J Chem Phys 61(5):2134–2138

    Article  CAS  Google Scholar 

  • Biehl R, Hoffmann B, Monkenbusch M, Falus P, Prevost S, Merkel R, Richter D (2008) Direct observation of correlated interdomain motion in alcohol dehydrogenase. Phys Rev Lett 101:138102-1–138102-4

    Article  Google Scholar 

  • Biswas R, Das AR, Pradhan T, Touraud D, Kunz W, Mahiuddin S (2008) Spectroscopic studies of catanionic reverse microemulsion: correlation with the superactivity of horseradish peroxidase enzyme in a restricted environment. J Phys Chem B 112(21):6620–6628

    Article  PubMed  CAS  Google Scholar 

  • Blum MM, Löhr F, Richardt A, Rüterjans H, Chen JCH (2006) Binding of a designed substrate analogue to diisopropyl fluorophosphatase: implications for the phosphotriesterase mechanism. J Am Chem Soc 128:12750–12757

    Article  PubMed  CAS  Google Scholar 

  • Blum MM, Mustyakimov M, Rüterjans H, Kehe K, Schoenborn BP, Langan P, Chen JCH (2009) Rapid determination of hydrogen positions and protonation states of diisopropyl fluorophosphatase by joint neutron and X-Ray diffraction refinement. PNAS 106:713–718

    Article  PubMed  CAS  Google Scholar 

  • Brûlet A, Lairez D, Lapp A, Cotton JP (2007) Improvement of data treatment in small-angle neutron scattering. J Appl Cryst 40:165–177

    Article  Google Scholar 

  • Bu Z, Biehl R, Monkenbusch M, Richter D, Callaway DJE (2005) Coupled protein domain motion in Taq polymerase revealed by neutron spin-echo spectroscopy. PNAS 102(49):17646–17651

    Article  PubMed  CAS  Google Scholar 

  • Burauer S, Sachert T, Sottmann T, Strey R (1999) On microemulsion phase behavior and monomeric solubility of surfactant. PCCP 1:4299–4306

    CAS  Google Scholar 

  • Burchard W, Richtering W (1989) Dynamic light scattering from polymer solutions. Progr Colloid Polymer Sci 80:151–163

    Article  CAS  Google Scholar 

  • Carnahan NF, Starling KE (1969) Equation of state for noninteracting rigid spheres. J Chem Phys 51(2):635–636

    Article  CAS  Google Scholar 

  • Coeur CL, Longeville S (2008) Microscopic protein diffusion at high concentration by neutron spin-echo spectroscopy. Chem Phys 345:298–304

    Article  Google Scholar 

  • Cotton JP (1991) Initial data treatment. In: Lindner P, Zemb T (eds), Neutron, X-ray and light scattering. Elsevier Science Publishers B.V.

  • Doster W, Longeville S (2007) Microscopic diffusion and hydrodynamic interactions of hemoglobin in red blood cells. Biophys J 93:1360–1368

    Article  PubMed  CAS  Google Scholar 

  • Endo H, Mihailescu M, Monkenbusch M, Allgaier J, Gompper G, Richter D, Jakobs B, Strey R, Grillo I (2001) Effect of amphiphilic block copolymers on the structure and phase behavior of oil-water-surfactant mixtures. J Chem Phys 115(1):580–600

    Article  CAS  Google Scholar 

  • Farago B, Gradzielski M (2001) The effect of the charge density of microemulsion droplets on the bending elasticity of their amphiphilic film. J Chem Phys 114(22):10105–10122

    Article  CAS  Google Scholar 

  • Farago B, Richter D, Huang JS, Safran SA, Milner ST (1990) Shape and size fluctuations of microemulsion droplets: the role of cosurfactant. Phys Rev Lett 65(26):3348–3351

    Article  PubMed  CAS  Google Scholar 

  • Farago B, Monkenbusch M, Goecking KD, Richter D, Huang JS (1995) Dynamics of microemulsions as seen by neutron spin echo. Physica B 213 & 214:712–717

    Article  Google Scholar 

  • Gäb J, Melzer M, Kehe K, Wellert S, Hellweg T, Blum MM (2010) Monitoring the hydrolysis of toxic organo-phosphates and -phosphonates by diisopropyl fluorophosphatase from Loligo vulgaris by 1D 1H-31P-HSQC NMR spectroscopy in aqueous solution and bicontinuous microemulsions. Anal Bioanal Chem 396:1213–1221

    Article  PubMed  Google Scholar 

  • Garcia Bernal JM, Garcia de la Torre J (1980) Transport properties and hydrodynamic centers of rigid macromolecules with arbitrary shapes. Biopolymers 19:751–766

    Article  CAS  Google Scholar 

  • Garcia de la Torre J, Bloomfield VA (1977) Hydrodynamic properties of macromolecular complexes. 1. Translation. Biopolymers 16:1747–1763

    Article  Google Scholar 

  • Garcia de la Torre J, Bloomfield VA (1981) Hydrodynamic properties of complex, rigid biological macromolecules: Theory and application. Quarterly Rev Biophys 14:81–139

    Article  CAS  Google Scholar 

  • Garcia de la Torre J, Huertas ML, Carrasco B (2000) Calculation of hydrodynamic properties of globular proteins from their atomic-level structure. Biophys J 78(2):719–730

    Article  PubMed  CAS  Google Scholar 

  • Glatter O, Orthaber D, Stradner A, Scherf G, Fanun M, Garti N, Clement V, Leser ME (2001) Sugar-ester nonionic microemulsion: structural characterization. J Coll Inter Sci 241:215–225

    Article  CAS  Google Scholar 

  • Hartleib J, Rüterjans H (2001) High-yield expression, purification, and characterization of the recombinant diisopropylfluorophosphatase from Loligo vulgaris. Prot Expr Purif 21(1):210–219

    Article  CAS  Google Scholar 

  • Helfrich W (1973) Elastic properties of lipid bilayers: theory and possible experiments. Z Naturforschung 28c:693–703

    Google Scholar 

  • Hellweg T (2002) Phase structures of microemulsions. Curr Opin Colloid Interface Sci 7:50–56

    Article  CAS  Google Scholar 

  • Hellweg T, von Klitzing R (2000) Structural changes and complex dynamics in the single phase region of a dodecane/C12E5/Water microemulsion: a dynamic light scattering study. Physica A 283(3–4):349–358

    Article  CAS  Google Scholar 

  • Hellweg T, Hinssen H, Eimer W (1993) A dynamic light scattering study on the two proteolytic fragments of gelsolin. Biophys J 65:799–805

    Article  PubMed  CAS  Google Scholar 

  • Hellweg T, Eimer W, Krahn E, Schneider K, Müller A (1997) Hydrodynamic properties of nitrogenase–the MoFe protein from Azotobacter vinelandii studied by dynamic light scattering and hydrodynamic modelling. Biochim Biophys Acta 1337:311–318

    Article  PubMed  CAS  Google Scholar 

  • Hellweg T, Brûlet A, Sottmann T (2000) Dynamics in an oil-continuous droplet microemulsions as seen by quasielastic scattering techniques. Phys Chem Chem Phys 2(22):5168–5174

    Article  CAS  Google Scholar 

  • Hellweg T, Gradzielski M, Farago B, Langevin D (2001) Shape fluctuation of microemulsion droplets: a neutron spin-echo study. Colloids Surfaces A 183–185:159–169

    Article  Google Scholar 

  • Hirai M, Hirai RK, Iwase H, Arai S, Mitsuya S, Takeda T, Seto H, Nagao M (1999) Dynamics of w/o AOT microemulsion studied by neutron spin echo. J Phys Chem Solids 60(8–9):1359–1361

    Article  CAS  Google Scholar 

  • Hirai M, Hirai RK, Iwase v, Hayakawa T, Kawabata Y, Takeda T (2002) Effect of proteins on dynamics of water-in-oil AOT microemulsion. Appl Phys A 74:S1254–S1256

    Article  CAS  Google Scholar 

  • Holderer O, Frielinghaus H, Byelov D, Monkenbusch M, Allgaier J, Richter D (2005) Dynamic properties of microemulsins modified with homopolymers and diblock copolymers: the determination of bending moduli and renormalization effects. J Chem Phys 122:094908/1–094908/8

    Article  CAS  Google Scholar 

  • Holmberg K (2001) Natural surfactants. Curr Opin Colloid Interface Sci 6:148–159

    Article  CAS  Google Scholar 

  • Hoskin FCG (1971) Diisopropylphosphorofluoridate and tabun: enzymatic hydrolysis and nerve function. Sci Agric 172:1243–1245

    CAS  Google Scholar 

  • Hoskin FCG, Roush AH (1982) Hydrolysis of nerve gas by squid-type diisopropyl phosphorofluoridate hydrolyzing enzyme on agarose resin. Sci Agric 215:1255–1257

    CAS  Google Scholar 

  • Huang JS, Milner ST, Farago B, Richter D (1987) Study of dynamics of microemulsion droplets by neutron spin-echo spectroscopy. Phys Rev Lett 59(22):2600–2603

    Article  PubMed  CAS  Google Scholar 

  • Iwanaga T, Suzuki M, Kunieda H (1998) Effect of added salts or polyols on the liquid crystalline structures of polyoxyethylene-type nonionic surfactants. Langmuir 14:5775–5781

    Article  CAS  Google Scholar 

  • Jahn W, Strey R (1988) Microstructure of microemulsions by freeze-fracture electron-microscopy. J Phys Chem 92:2294–2301, Keine Kopie

    Google Scholar 

  • Kahlweit M et al (1987) How to study microemulsions. J Colloid Interface Sci 118(2):436–453

    Article  CAS  Google Scholar 

  • Kahlweit M, Strey R (1985) Phasenverhalten ternärer Systeme des Typs H2O-Öl-nichtionisches Amphiphil (Mikroemulsionen). Angew Chem 97:655–669

    Article  CAS  Google Scholar 

  • Kawabata Y, Seto H, Nagao M, Takeda T (2007) Pressure effects on bending elasticities of surfactant monolayers in a ternary microemulsion composed of aerosol-OT/D2O/decane. J Chem Phys 127:044705-1–044705-9

    Article  Google Scholar 

  • Kirkwood JG (1949) The statistical mechanical theory of irreversible processes in solutions of flexible macromolecules. Recueil Trav Chim 68:649–660

    Article  CAS  Google Scholar 

  • Koeper I, Bellissent-Funel MC (2000) Hindered protein dynamics in the presence of a cryoprotecting agent. Appl Phys A 74:S1257–S1259

    Article  Google Scholar 

  • Koepke J, Scharff EI, Lücke C (2003) Statistical analysis of crystallographic data obtained from squid ganglion DFPase at 0.85 Å resolution. Acta Crystallographica D 59:1744–1754

    Google Scholar 

  • Kohlbrecher J (2008) SASfit: A program for fitting simple structural models to small angle scattering data. Paul Scherrer Institut, Laboratory for Neutron Scattering, CH-5232 Villigen, Switzerland

  • Komves C, Osborne D, Russell AJ (1994) Degredation of pesticides in a contiuous-flow 2-phase microemulsion reactor. Biotechnol Prog 10(3):340–343

    Article  Google Scholar 

  • Koppel DE (1972) Analysis of macromolecular polydispersity in intensity correlation spectroscopy: the method of cumulants. J Chem Phys 57(11):4814–4820

    Article  CAS  Google Scholar 

  • Larsson KM, Adlercreutz P, Mattiasson B, Olsson U (1991) Enzyme catalysis in uni- and bi-continuous microemulsions: dependence of kinetics on substrate partitioning. J Chem Soc Faraday Trans 87:465–471

    Article  CAS  Google Scholar 

  • Lee KM, Biellmann JF (1987) Enzyme and organic solvents: horse liver alcohol dehydrogenase in non-ionic microemulsion: stability and activity. FEBS Lett 223(1):33–36

    Article  PubMed  CAS  Google Scholar 

  • Lutter S, Koetz J, Tiersch B, Kosmella S (2009) Polymer-modified bicontinuous microemulsions used as a template for the formation of nanorods. J Dispersion Sci Technol 30:745–752

    Article  CAS  Google Scholar 

  • Mihailescu M, Monkenbusch M, Endo H, Allgaier J, Gompper G, Richter D, Jakobs B, Sottmann T, Farago B (2001) Dynamics of bicontinuous microemulsion phases with and without amphiphilic block-copolymers. J Chem Phys 115(20):9563–9577

    Article  CAS  Google Scholar 

  • Milner ST, Safran SA (1987) Dynamical fluctuations of droplet microemulsions and vesicles. Phys Rev A 36(9):4371–4379

    Article  PubMed  CAS  Google Scholar 

  • Möller A, Lang P, Findenegg GH, Keiderling U (1998) Location of butanol in mixed micelles with alkyl glucosides studied by SANS. J Phys Chem B 102(45):8958–8964

    Article  Google Scholar 

  • Monkenbusch M (1997) The Jülich neutron spin echo spectrometer. Neutron News 8(1):25

    Article  Google Scholar 

  • Monkenbusch M, Schatzler R, Richter D (1997) The Julich neutron spin-echo spectrometer–design and performanc. Nucl Instrum Methods Phys Res Sect A 399(2–3):301–323

    Article  CAS  Google Scholar 

  • Pedersen JS (1997) Analysis of small-angle scattering data from colloids and polymer solutions: modeling and least-square fitting. Adv Coll Inter Sci 70:171–210

    Article  CAS  Google Scholar 

  • Provencher SW (1982a) A constrained regularization method for inverting data represented by linear algebraic or integral equations. Comput Phys Com 27:213–217

    Article  Google Scholar 

  • Provencher SW (1982b) Contin: a general purpose constrained regularization program for inverting noisy linear algebraic and integral equations. Comput Phys Com 27:229–242

    Article  Google Scholar 

  • Richardt A, Blum MM (2008) Decontamination of Warfare Agents Wiley-VCH, Weinheim ISBN-10:3-527-31756-2

  • Riseman J, Kirkwood JG (1950) The intrinsic viscosity, translational and rotatory diffusion constants of rod-like macromolecules in solution. J Chem Phys 18(4):512–516

    Article  CAS  Google Scholar 

  • Rojas O, Koetz J, Kosmella S, Tiersch B, Kramer M, Wacker P (2009) Structural studies of ionic liquid-modified microemulsions. J Coll Interf Sci 33:782–790

    Article  Google Scholar 

  • Safran SA (1999) Curvature elasticity of thin films. Adv Phys 48(4):395–448 Review

    Google Scholar 

  • Scharff EI, Koepke J, Fritzsch G, Lücke C, Rüterjans H (2001) Crystal structure of diisopropylfluorophosphatase from Loligo vulgaris. Structure 9:493–502

    Article  PubMed  CAS  Google Scholar 

  • Sottmann T, Strey R (1997) Ultralow interfacial tensions in water-n-alkane-surfactant systems. J Chem Phys 106(20):8606–8615

    Article  CAS  Google Scholar 

  • Stamatis H, Xenakis A, Kolisis FN (1999) Bioorganic reactions in microemulsions: the case of lipases. Biotechnl Adv 17:293–318

    Article  CAS  Google Scholar 

  • Stradner A, Glatter O, Schurtenberger P (2000) A hexanol-induced sphere-toflexible cylinder transition in aqueous alkyl polyglucoside solutions. Langmuir 16(12):5354–5364

    Article  CAS  Google Scholar 

  • Strey R (1994) Microemulsion microstructure and interfacial curvature. Coll Polymer Sci 272:1005–1019

    Article  CAS  Google Scholar 

  • Strey R (1996) Phase behavior and interfacial curvature in water-oil-surfactant systems. Curr Opin Colloid Interface Sci 1:402–410

    Article  CAS  Google Scholar 

  • Stubenrauch C (2001) Sugar surfactants–aggregation, interfacial, and adsorption phenomena. Curr Opin Colloid Interface Sci 6:160–170

    Article  CAS  Google Scholar 

  • Teubner M, Strey R (1987) Origin of the scattering peak in microemulsions. J Chem Phys 87(5):3195–3200

    Article  CAS  Google Scholar 

  • Wellert S, Karg M, Imhof H, Steppin A, Altmann HJ, Dolle M, Richardt A, Tiersch B, Koetz J, Lapp A, Hellweg T (2008) Structure of Biodiesel based bicontinuous microemulsions for environmentally compatible decontamination as seen by small angle neutron scattering and freeze fracture electron microscopy. J Colloid Interf Sci 325:250–258

    Article  CAS  Google Scholar 

  • Wood K, Caronna C, Fouquet P, Häussler W, Natali F, Ollivier J, Orecchini A, Plazanet M, Zaccai G (2008) A benchmark for protein dynamics: Ribonuclease as measured by neutron scattering in a large wavevector-energy transfer range. Chem Phys 345:305–314

    Article  CAS  Google Scholar 

  • Zilman AG, Granek R (1996) Undulations and dynamic structure factor of membranes. Phys Rev Lett 77:4788–4791

    Article  PubMed  CAS  Google Scholar 

  • Zouni A, Kern J, Frank J, Hellweg T, Behlke J, Saenger W, Irrgang KD (2005) Size determination of cyanobacterial and higher plant photosystem II by gel permeation chromatography, light scattering, and ultracentrifugation. Biochem 44(11):4572–4581

    Article  CAS  Google Scholar 

Download references

Acknowledgments

J.G. and M.M.B. were supported by the German Ministry of Defense under contract E/UR3G/6G115/6A801. T.H., R.S., C.S. also were financed by the German Ministry of Defense (Contracts E/E590/9Z004/5F159 and E/E590/8Z002/4F170). The SANS experiments were supported by the European Union through the NMI3 program. We are grateful to the Laboratoire Léon Brillouin and the Jülich Center for Neutron Scattering at the FRM II for providing beamtime.

Author information

Author notes

  1. Jürgen Gäb

    Present address: Institute of Pharmaceutical Chemistry, Phillipps University Marburg, 35032, Marburg, Germany

  2. Marc-Michael Blum

    Present address: Bioscience Division, Los Alamos National Laboratory, PO Box 1663, Mailstop G758, Los Alamos, NM, 87545, USA

Authors and Affiliations

  1. TU Berlin, Institut für Chemie, Stranski-Laboratorium für Physikalische und Theoretische Chemie, Straße des 17.Juni 124, 10623, Berlin, Germany

    Stefan Wellert

  2. Institut für Chemie, Universität Potsdam, 14469, Potsdam, Germany

    Brigtte Tiersch & Joachim Koetz

  3. Wehrwissenschaftliches Institut für Schutztechnologien–ABC-Schutz, Humboldtstraße, 29633, Munster, Germany

    André Richardt

  4. Laboratoire Léon Brillouin, C.E.A.-C.E.N. Saclay, 91191, Gif-sur-Yvette Cedex, France

    Alain Lapp

  5. Jülich Center for Neutron Scattering, FRM II Garching, Lichtenbergstr. 1, 85747, Garching, Germany

    Olaf Holderer

  6. Blum-Scientific Services, Cecilienstrasse 3, 22301, Hamburg, Germany

    Jürgen Gäb & Marc-Michael Blum

  7. Soft Matter und Funktionale Materialien, Helmholtz-Zentrum Berlin für Materialien und Energie, Hahn-Meitner Platz1, 14109, Berlin, Germany

    Ralf Stehle

  8. Physikalische und Biophysikalische Chemie, Universitätsstr. 25, 33615, Bielefeld, Germany

    Christoph Schulreich & Thomas Hellweg

Authors
  1. Stefan Wellert
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Brigtte Tiersch
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Joachim Koetz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. André Richardt
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Alain Lapp
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Olaf Holderer
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jürgen Gäb
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Marc-Michael Blum
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Christoph Schulreich
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Ralf Stehle
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Thomas Hellweg
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Stefan Wellert or Thomas Hellweg.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wellert, S., Tiersch, B., Koetz, J. et al. The DFPase from Loligo vulgaris in sugar surfactant-based bicontinuous microemulsions: structure, dynamics, and enzyme activity. Eur Biophys J 40, 761–774 (2011). https://doi.org/10.1007/s00249-011-0689-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00249-011-0689-0

Keywords

Search

Navigation