Skip to main content
SpringerLink
Log in

Permeability and reactivity of Thermotoga maritima in latex bimodal blend coatings at 80°C: a model high temperature biocatalytic coating

  • Original Paper
  • Published:
Extremophiles Aims and scope Submit manuscript

Abstract

Thermostable polymers cast as thin, porous coatings or membranes may be useful for concentrating and stabilizing hyperthermophilic microorganisms as biocatalysts. Hydrogel matricies can be unstable above 65°C. Therefore a 55-μm thick, two layer (cell coat + polymer top coat) bimodal, adhesive latex coating of partially coalesced polystyrene particles was investigated at 80°C using Thermotoga maritima as a model hyperthermophile. Coating permeability (pore structure) was critical for maintaining T. maritima viability. The permeability of bimodal coatings generated from 0.8 v/v of a suspension of non-film-forming 800 nm polystyrene particles with high glass transition temperature (Tg= 94°C, 26.9% total solids) blended with 0.2 v/v of a suspension of film-forming 158 nm polyacrylate/styrene particles (Tg≈ −5°C, 40.9% total solids) with 0.3 g sucrose/g latex was measured in a KNO3 diffusion cell. Diffusivity ratio remained above 0.04 (Deff/D) when incubated at 80°C in artificial seawater (ASW) for 5 days. KNO3 permeability was corroborated by cryogenic-SEM images of the pore structure. In contrast, the permeability of a mono-dispersed acrylate/vinyl acetate latex Rovace SF091 (Tg~10°C) rapidly decreased and became impermeable after 2 days incubation in ASW at 80°C. Thermotoga maritima were entrapped in these coatings at a cell density of 49 g cell wet weight/liter of coating volume, 25-fold higher than the density in liquid culture. Viable T. maritima were released from single-layer coatings at 80°C but accurate measurement of the percentage of viable entrapped cells by plate counting was not successful. Metabolic activity could be measured in bilayer coatings by utilization of glucose and maltose, which was identical for latex-entrapped and suspended cells. Starch was hydrolyzed for 200 h by latex-entrapped cells due to the slow diffusion of starch through the polymer top coat compared to only 24 h by suspended T. maritima. The observed reactivity and stability of these coatings was surprising since cryo-SEM images suggested that the smaller low Tg polyacrylate/styrene particles preferentially bound to the T. maritima toga-sheath during coat formation. This model system may be useful for concentrating, entrapment and stabilization of metabolically active hyperthermophiles at 80°C.

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

Similar content being viewed by others

Abbreviations

ASW:

Artificial seawater

Deff/D:

Ratio of the diffusivity of KNO3 in a latex coating relative to its diffusivity, D, in water

T g :

Polymer particle glass transition temperature

v/v:

Volume fraction of bimodal latex emulsions blended

References

  • Adams MWW (1995) Large-scale growth of hyperthermophiles. In: Robb FT, Place AR, Sowers KR, Schrier HJ, DasSarma S, Fleischmann EM (eds) Archaea: a laboratory manual. Cold Spring Harbor, New York, pp 47–49

    Google Scholar 

  • Brown SH, Sjrholm C, Kelly RM (1993) Purification and characterization of a highly stable glucose isomerase produced by the extremely thermophilic eubacterium, Thermotoga maritima. Biotechnol Bioeng 41:878–886

    Google Scholar 

  • Cabral JB, Mota M, Tramper J (2001) Multiphase bioreactor design. Tayor & Francis, London

    Google Scholar 

  • Cantwell JB, Mills PDA, Jones E, Stewart RF (1995) Immobilized cells. European Patent Office 0288203B1

  • Charaniya SP (2004) Optimization of the catalytic activity of bilayer latex coatings for bacterial whole-cell oxidation and reduction. MS Thesis, University of Minnesota

  • Chhabra SR, Shockley KR, Conners SB, Scott KL, Wolfinger RD, Kelly RM (2003) Carbohydrate-induced differential gene expression patterns in the hyperthermophilic bacterium Thermotoga maritima. J Biol Chem 278:7540–7552

    Google Scholar 

  • Cussler EL (1998) Diffusion: mass transfer in fluid systems, 2nd edn. Cambridge University Press, Cambridge, pp 22–24

    Google Scholar 

  • De Rosa M, Gambacorta A, Lama L, Nicolaus B (1981) Immobilization of thermophilic microbial cells in crude egg white. Biotechnol Lett 3:183–186

    Google Scholar 

  • Di Lernia I, Schiraldi C, Generoso M, De Rosa M (2002) Trehalose production at high temperature exploiting an immobilized cell bioreactor. Extremophiles 6:341–347

    Google Scholar 

  • Dos Santos FD, Fabre P, Drujon X, Meunier G, Leibler L (2000) Films from soft-core/hard-shell hydrophobic latexes: structure and thermomechanical properties. J Polymer Sci B 38:2989–3000

    Google Scholar 

  • Drioli E, Giorno L (1999) Biocatalytic membrane reactors. Taylor & Francis, London

    Google Scholar 

  • Geankoplis CJ (1984) Mass transport phenomena. Edward Brothers, Columbus

    Google Scholar 

  • Gebhard MS, Lesko PM, Brown AB, Young DH (2004) Porous non-friable polymer films. US Patent 6,750,050

  • Hicks PM, Kelly RM (1999) Thermophilic microorganisms. In: Flickinger MC, Drew SW (eds) Encyclopedia of bioprocess technology: fermentation, biocatalysis, and bioseparation. Wiley Interscience, New York, pp 2536–2552

    Google Scholar 

  • Holst O, Manelius Å, Krahe M, Märkl H, Raven N, Sharp R (1997) Thermophiles and fermentation technology. Comp Biochem Physiol 118A:415–422

    Google Scholar 

  • Huang Z, Thiagarajan V, Lyngberg OK, Scriven LE, Flickinger MC (1999) Microstructure evolution in polymer latex coatings for whole-cell biocatalyst applications. J Coll Interface Sci 215:226–243

    Google Scholar 

  • Jons S, Ries P, McDonald CJ (1999) Porous latex composite membranes: fabrication and properties. J Membr Sci 155:79–99

    Google Scholar 

  • Kalinina O, Kumacheva E (1999) A “core-shell” approach to producing 3D polymer nanocomposites. Macromolecules 32:4122–4129

    Google Scholar 

  • Kester DR, Duedall IW, Connors DN, Pyrkowicz RM (1967) Preparation of artificial sea water. Limnol Oceanogr 12:176–178

    Google Scholar 

  • Liebl W, Stemplinger I, Ruile P (1997) Properties and gene structure of the Thermotoga maritima α-amylase AmyA, a putative lipoprotein of a hyperthermophilic bacterium. J Bacteriol 179:941–948

    Google Scholar 

  • Lyngberg OK, Thiagaragan V, Stemke DJ, Schottel JL, Scriven LE, Flickinger MC (1999a) A patch coating method for preparing biocatalytic films with E. coli. Biotechnol Bioeng 62:44–55

    Google Scholar 

  • Lyngberg OK, Stemke DJ, Schottel JL, Flickinger MC (1999b) A simple single use luciferase based mercury biosensor using latex-film immobilized Escherichia coli HB101. J Ind Microbiol Biotechnol 23:668–676

    Google Scholar 

  • Lyngberg OK, Ng CP, Thiagarajan V, Scriven LE, Flickinger MC (2001) Engineering the microstructure and permeability of thin multilayer latex biocatalytic coatings containing E. coli. Biotechnol Prog 17:1169–1179

    Google Scholar 

  • Ma Y (2002) High-resolution cryo-scanning electron microscopy of latex film formation. PhD Thesis, University of Minnesota

  • Pantazaki AA, Pritsa AA, Kyriakidis DA (2002) Biologically relevant enzymes from Thermus thermophilus. Appl Microbiol Biotechnol 58:1–12

    Article  Google Scholar 

  • Park H-S, Kayser KJ, Kwak J-H, Kilbane JJ (2004) Heterologous gene expression in Thermus thermophilus: β-galactosidase, dibenzothiophene monooxygenase, PNB carboxy esterase, 2-aminobiphenyl-2,3-diol dioxygenase, and chloramphenicol acetyl transferase. J Ind Microbiol Biotechnol 31:189–197

    Google Scholar 

  • Pörtner R, Märkl H (1996) Immobilization of the extremely thermophilic archaeon Pyrococcus furiosus in macro-porous carriers. In: Wijffels RH, Buitelaar RM, Bucke C, Tramper J (eds) Immobilized cells: basics and applications. Elsevier, Amsterdam, pp 424–430

    Google Scholar 

  • Rainina EI, Pusheva MA, Ryabokon AM, Bolotina NP, Lozinsky VI, Varfolomeyev SD (1994) Microbial cells immobilized in poly(vinyl alcohol) cryogels: biocatalytic reduction of CO2 by the thermophilic homoacetogenic bacterium Acetogenium kiuvi Biotechnol Appl Biochem 19:321–329

    Google Scholar 

  • Ryabokon AM, Kevbrina MV, Pusheva MA, Zubov AL, Lozinsky VI, Rainina EI (1996) Ecologically pure cultures of acetate synthesis on diverse gaseous substrates by homoacetogenic bacteria, entrapped in poly(vinyl alcohol) cryogel. In: Wijffels RH, Buitelaar RM, Bucke C, Tramper J (eds) Immobilized cells: basics and applications. Elsevier, Amsterdam, pp 106–111

    Google Scholar 

  • Sanchez Marcano JG, Tsotsis TT (2002) Catalytic membranes and membrane reactors. Wiley-VCH, Weinheim, Germany

    Google Scholar 

  • Schiraldi C, Di Lernia I, Giuliano M, Generoso M, D’ Agnostino A, De Rosa M (2003) Evaluation of a high temperature immobilized enzyme reactor for production of non-reducing oligosaccharides. J Ind Microbiol Biotechnol 30:302–307

    Google Scholar 

  • Schröder C, Selig M, Schönheit P (1994) Glucose fermentation to acetate, CO2, and H2 in the anaerobic hyperthermophilic eubacterium Thermotoga maritima: involvement of the Embden-Mayerhof pathway. Arch Microbiol 161:460–470

    Google Scholar 

  • Sharp RJ, Raven NDH (1997) Isolation and growth of hyperthermophiles. In: Rhodes PM, Standbury PF (eds) Applied microbial physiology. IRL, Oxford, pp 23–52

    Google Scholar 

  • Singh A, Goel R, Johri BN (1990) Production of cellulolytic enzymes by immobilized Sporotrichium thermophile. Enz Microbiol Technol 12:464–468

    Google Scholar 

  • Solheid C (2003) Characterization of bilayer latex coatings containing viable Gluconobacter oxydans for the oxidation of D-sorbitol to L-sorbose. MS Thesis, University of Minnesota

  • Swope KL, Flickinger MC (1996) Activation and regeneration of whole cell biocatalysts: initial and periodic induction behavior in starved E. coli after immobilization in thin synthetic films. Biotechnol Bioeng 51:360–370

    Google Scholar 

  • Thiagarajan VS, Huang Z, Scriven LE, Schottel JL, Flickinger MC (1999) Microstructure of a biocatalytic latex coating containing viable Escherichia coli cells. J Coll Interface Sci 215:244–257

    Google Scholar 

  • Tzitznou A, Keddie JL (2000) Film formation of latex blends with bimodal particle size distributions: consideration of particle deformability and continuity of the dispersed phase. Macromolecules 33:2695–2708

    Google Scholar 

  • Tzitznou A, Keddie JL, Geurts JM, Peters ACIA, Satguru R (2000) Film formation of latex blends with bimodal particle size distributions: consideration of particle deformability and continuity of the dispersed phase. Macromol 33:2695–2708

    Google Scholar 

  • Van de Ban E, Willemen H, Wassink H, Haaker H, Laane C (1998) Bioreductions by Pyrococcus furiosus at elevated temperature. In: Ballesteros A, Plou FJ, Iborra JL, Halling PJ (eds) Progress in biotechnology 15: stability and stabilization of biocatalysts. Elsevier, Amsterdam

    Google Scholar 

  • Van Niel EWJ, Claassen PAM, Stams AJM (2003) Substrate and product inhibition of hydrogen production by the extreme thermophile Caldicellulosiruptor saccharolyticus. Biotechnol Bioeng 81:255–262

    Article  PubMed  Google Scholar 

  • Van Ooteghem SA, Beer SK, Yue PC (2002) Hydrogen production by the thermophilic bacterium Thermotoga neapolitana. Appl Biochem Biotechnol 98–100:177–189

    Google Scholar 

  • Webb C, Devakos GA (1996) Studies in viable cell immobilization. Academic Press, RG Landes Co, Austin

    Google Scholar 

  • Wijffels RH (ed) (2001) Immobilized cells. Springer, Berlin

    Google Scholar 

  • Worden RM, Subramanian R, Bly MJ, Winter S, Aronson CL (1991) Growth kinetics of Bacillus stearothermophilus BR219. Appl Biochem Biotechnol 28/29:267–275

    Google Scholar 

Download references

Acknowledgements

The authors would like to thank Professor Robert M. Kelly, Tina D. Rinker, and Paula M. Hicks, North Carolina State University, Raleigh, North Carolina, for their assistance and encouragement in beginning this work, Sridevi Nagarajan for developing the anaerobic plate counting method, Erwin Sutanto for cryo-FESEM images of bimodal blend coatings, and Marcello Fidaleo for helpful comments on preparation of the manuscript. Latex polymers were generously supplied by Matthew Gebhard, Rohm and Haas, Co., Spring House, PA, USA. This work was supported by NIH grant T32/GM08347, DSO DARPA contract N66001-020C-8046, Joe Bielitski Program Director, and the University of Minnesota BioTechnology Institute.

Author information

Author notes

  1. Olav K. Lyngberg

    Present address: Bristol-Myers Squibb, One Squibb Drive, New Brunswick, NJ, 08903-0191, USA

  2. Yue Ma

    Present address: Intel Corporation, 2501 NW 229th Street, Hillsboro, OR, 97124, USA

Authors and Affiliations

  1. Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, MN, 55455, USA

    Olav K. Lyngberg, Yue Ma & L. E. Scriven

  2. Biotechnology Institute, University of Minnesota, 140 Gortner Laboratories, Saint Paul, MN, 55108-6106, USA

    Olav K. Lyngberg, Chris Solheid, Salim Charaniya, Venkata Thiagarajan & Michael C. Flickinger

  3. Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Saint Paul, MN, 55108-6106, USA

    Michael C. Flickinger

Authors
  1. Olav K. Lyngberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Chris Solheid
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Salim Charaniya
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Yue Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Venkata Thiagarajan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. L. E. Scriven
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Michael C. Flickinger
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Michael C. Flickinger.

Additional information

Communicated by J. Wiegel

Rights and permissions

Reprints and permissions

About this article

Cite this article

Lyngberg, O.K., Solheid, C., Charaniya, S. et al. Permeability and reactivity of Thermotoga maritima in latex bimodal blend coatings at 80°C: a model high temperature biocatalytic coating. Extremophiles 9, 197–207 (2005). https://doi.org/10.1007/s00792-005-0434-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00792-005-0434-7

Keywords

Search

Navigation