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Isolation and characterization of a new cellulosome-producing Clostridium thermocellum strain

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Abstract

The anaerobic thermophilic bacterium, Clostridium thermocellum, is a potent cellulolytic microorganism that produces large extracellular multienzyme complexes called cellulosomes. To isolate C. thermocellum organisms that possess effective cellulose-degrading ability, new thermophilic cellulolytic strains were screened from more than 800 samples obtained mainly from agriculture residues in Thailand using microcrystalline cellulose as a carbon source. A new strain, C. thermocellum S14, having high cellulose-degrading ability was isolated from bagasse paper sludge. Cellulosomes prepared from S14 demonstrated faster degradation of microcrystalline cellulose, and 3.4- and 5.6-fold greater Avicelase activity than those from C. thermocellum ATCC27405 and JW20 (ATCC31449), respectively. Scanning electron microscopic analysis showed that S14 had unique cell surface features with few protuberances in contrast to the type strains. In addition, the cellulosome of S14 was resistant to inhibition by cellobiose that is a major end product of cellulose hydrolysis. Saccharification tests conducted using rice straw soaked with sodium hydroxide indicated the cellulosome of S14 released approximately 1.5-fold more total sugars compared to that of ATCC27405. This newly isolated S14 strain has the potential as an enzyme resource for effective lignocellulose degradation.

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References

  • Bayer EA, Lamed R (1986) Ultrastructure of the cell surface cellulosome of Clostridium thermocellum and its interaction with cellulose. J Bacteriol 167:828–836

    PubMed  CAS  Google Scholar 

  • Bayer EA, Kenig R, Lamed R (1983) Adherence of Clostridium thermocellum to cellulose. J Bacteriol 156:818–827

    PubMed  CAS  Google Scholar 

  • Bayer EA, Shoham Y, Lamed R (2008) Cellulosome-enhanced conversion of biomass: on the road to bioethanol. In: Wall JD, Harwood CS, Demain AL (eds) Bioenergy. ASM Press, Washington, DC, pp 75–96

    Google Scholar 

  • Béguin P, Millet J, Aubert JP (1992) Cellulose degradation by Clostridium thermocellum: from manure to molecular biology. FEMS Microbiol Lett 9:523–528

    Article  Google Scholar 

  • Bensadoun A, Weinstein D (1976) Assay of proteins in the presence of interfering materials. Anal Biochem 70:241–250

    Article  PubMed  CAS  Google Scholar 

  • Brown SD, Raman B, McKeown CK, Kale SP, He Z, Mielenz JR (2007) Construction and evaluation of a Clostridium thermocellum ATCC 27405 whole-genome oligonucleotide microarray. Appl Biochem Biotechnol 137–140:663–674

    Article  PubMed  Google Scholar 

  • Dashtban M, Schraft H, Qin W (2009) Fungal bioconversion of lignocellulosic residues; opportunities & perspectives. Int J Biol Sci 5:578–595

    Article  PubMed  CAS  Google Scholar 

  • Demain AL, Newcomb M, David JH (2005) Cellulase, Clostridia, and Ethanol. Microbiol Mol Biol Rev 69:124–154

    Article  PubMed  CAS  Google Scholar 

  • Freier D, Mothershed CP, Wiegel J (1988) Characterization of Clostridium thermocellum JW20. Appl Environ Microbiol 54:204–211

    PubMed  CAS  Google Scholar 

  • Fujino T, Béguin P, Aubert JP (1993) Organization of a Clostridium thermocellum gene cluster encoding the cellulosomal scaffolding protein CipA and a protein possibly involved in attachment of the cellulosome to the cell surface. J Bacteriol 175:1891–1899

    PubMed  CAS  Google Scholar 

  • Gerhardt P, Murray RGE, Costilow RN, Nester EW, Wood WA, Krieg NR, Phillips GB (1981) Manual of methods for general bacteriology. American Society of Microbiology, Washington, DC

    Google Scholar 

  • Hungate RE (1969) A roll-tube method for cultivation of strict anaerobes. Methods Microbiol 3:117–132

    Article  Google Scholar 

  • Johnson EA, Reese T, Demain AL (1982a) Inhibition of Clostridium thermocellum cellulase by end products of cellulolysis. J Appl Biochem 4:64–71

    CAS  Google Scholar 

  • Johnson EA, Sakajoh M, Halliwell G, Media A, Demain AL (1982b) Saccharification of complex cellulosic substrates by the cellulase system from Clostridium thermocellum. Appl Environ Microbiol 43:1125–1132

    PubMed  CAS  Google Scholar 

  • Lamed R, Setter E, Kenig R, Bayer EA (1983) The cellulosome-a discrete cell surface organelle of Clostridium thermocellum which exhibits separate antigenic, cellulose-binding and various cellulolytic activities. Biotechnol Bioeng Symp 13:163–181

    CAS  Google Scholar 

  • Ljungdahl LG, Coughlan MP, Mayer F, Mori Y, Hon-nami H, Hon-nami K (1988) Macrocellulase complexes and yellow affinity substance from Clostridium thermocellum. In: Wood WA, Kellogg ST (eds) Methods in enzymology, vol 160. Academic Press, New York, pp 483–500

    Google Scholar 

  • Lynd LR, Weimer PJ, van Zyl WH, Pretorius IS (2002) Microbial cellulose utilization: fundamentals and biotechnology. Microbiol Mol Biol Rev 66:506–577

    Article  PubMed  CAS  Google Scholar 

  • Mayer F, Coughlan MP, Mori Y, Ljungdahl L (1987) Macromolecular organization of the cellulolytic enzyme complex of Closridium thermocellum as revealed by electron microscopy. Appl Eviron Microbiol 53:2785–2792

    CAS  Google Scholar 

  • McCann MC, Carpita NC (2008) Designing the deconstruction of plant cell walls. Curr Opin Plant Biol 11:314–320

    Article  PubMed  CAS  Google Scholar 

  • Morag E, Bayer EA, Lamed R (1992) Affinity digestion for the near-total recovery of purified cellulosome from Clostridium thermocellum. Enzyme Microb Technol 14:289–292

    Article  CAS  Google Scholar 

  • Mori Y (1992) Comparison of the Cellulolytic Systems of Clostridium thermocellum YM4 and JW20. Biotechnol Lett 14:131–136

    Article  CAS  Google Scholar 

  • Nelson N (1944) A photometric adaptation of the Somogyi method for the determination of glucose. J Biol Chem 153:375–380

    CAS  Google Scholar 

  • Ng TK, Weimer PJ, Zeikus JG (1977) Cellulolytic and physiological properties of Clostridium thermocellum. Arch Microbiol 114:1–7

    Article  PubMed  CAS  Google Scholar 

  • Ohmiya K, Sakka K, Kimura T, Morimoto K (2003) Application of microbial genes to recalcitrant biomass utilization and environmental conservation. J Biosci Bioeng 95:549–561

    PubMed  CAS  Google Scholar 

  • Ozkan M, Desai SG, Zhang Y, Stevenson DM, Beane J, White EA, Guerinot ML, Lynd LR (2001) Characterization of 13 newly isolated strains of anaerobic, cellulolytic, thermophilic bacteria. J Ind Microbiol Biotechnol 27:275–280

    Article  PubMed  CAS  Google Scholar 

  • Raman B, Pan C, Hurst GB, Rodriguez M Jr, McKeown CK, Lankford PK, Samatova NF, Mielenz JR (2009) Impact of pretreated Switchgrass and biomass carbohydrates on Clostridium thermocellum ATCC 27405 cellulosome composition: a quantitative proteomic analysis. PLoS One 4:e5271

    Article  PubMed  Google Scholar 

  • Schwarz WH (2001) The cellulosome and cellulose degradation by anaerobic bacteria. Appl Microbiol Biotechnol 56:634–649

    Article  PubMed  CAS  Google Scholar 

  • Sukhumavasi J, Ohmiya K, Shimizu S, Ueno K (1988) Clostridium Jousi sp. nov., a cellulolytic, moderate thermophilic species from Thai compost. Int J Syst Bacteriol 38:179–182

    Article  Google Scholar 

  • Sun Y, Cheng J (2002) Hydrolysis of lignocellulosic materials for ethanol production: a review. Bioresour Technol 83:1–11

    Article  PubMed  CAS  Google Scholar 

  • Viljoen JA, Fred EB, Peterson WH (1926) The fermentation of cellulose by thermophilic bacteria. J Agric Sci 16:1–17

    CAS  Google Scholar 

  • Virunanon C, Chantaroopamai S, Denduangbaripant J, Chulalaksananukul W (2008) Solventogenic-cellulolytic Clostridia from 4-step-screening process in agricultural waste and cow intestinal tract. Anaerobe 14:109–117

    Article  PubMed  CAS  Google Scholar 

  • Wood WA, Kellogg ST (1988) Methods in enzymology, vol 160. Academic Press, New York

    Google Scholar 

Download references

Acknowledgments

C. Tachaapaikoon was supported by the Visiting Research Fellowship Program of Japan International Research Center for Agricultural Sciences (JIRCAS) and Thailand Research Fund (TRF Grant). This work was conducted as part of a research project funded by a grant (Development of Biomass Utilization Technologies for Revitalizing Rural Areas) from the Ministry of Agriculture, Forestry, and Fisheries of Japan.

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Authors and Affiliations

  1. Post-harvest Science and Technology Division, Japan International Research Center for Agricultural Sciences (JIRCAS), 1-1 Ohwashi, Tsukuba, Ibaraki, 305-8686, Japan

    Chakrit Tachaapaikoon, Akihiko Kosugi, Rattiya Waeonukul, Takamitsu Arai, Yoshinori Murata & Yutaka Mori

  2. Pilot Plant Development and Training Institute (PDTI), King Mongkut’s University of Technology Thonburi (KMUTT), Bangkok, Thailand

    Chakrit Tachaapaikoon & Patthra Pason

  3. School of Bioresources and Technology, King Mongkut’s University of Technology, Thonburi (KMUTT), Bangkok, Thailand

    Rattiya Waeonukul, Khanok Ratanakhanokchai & Khin Lay Kyu

Authors
  1. Chakrit Tachaapaikoon
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  2. Akihiko Kosugi
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  3. Patthra Pason
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  4. Rattiya Waeonukul
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  5. Khanok Ratanakhanokchai
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  6. Khin Lay Kyu
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  7. Takamitsu Arai
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  8. Yoshinori Murata
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  9. Yutaka Mori
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Corresponding author

Correspondence to Yutaka Mori.

Additional information

Chakrit Tachaapaikoon and Akihiko Kosugi contributed equally to this work.

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Tachaapaikoon, C., Kosugi, A., Pason, P. et al. Isolation and characterization of a new cellulosome-producing Clostridium thermocellum strain. Biodegradation 23, 57–68 (2012). https://doi.org/10.1007/s10532-011-9486-9

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  • DOI: https://doi.org/10.1007/s10532-011-9486-9

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