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Cellulase Assays

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Biofuels

Part of the book series: Methods in Molecular Biology ((MIMB,volume 581))

Summary

Cellulose is a heterogeneous polysaccharide, and its enzymatic hydrolysis requires endoglucanase, exoglucanase (cellobiohydrolase), and β-glucosidase to work together. We summarize the most commonly used assays for individual enzymes and cellulase mixture.

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References

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

    Article  CAS  Google Scholar 

  2. Zhang Y-HP, Himmel M, and Mielenz JR (2006) Outlook for cellulase improvement: screening and selection strategies. Biotechnol. Adv. 24(5):452–481.

    Article  CAS  Google Scholar 

  3. Zhang Y-HP and Lynd LR (2004) Toward an aggregated understanding of enzymatic hydrolysis of cellulose: noncomplexed cellulase systems. Biotechnol. Bioeng. 88:797–824.

    Article  CAS  Google Scholar 

  4. Zhang Y-HP (2008) Reviving the carbohydrate economy via multi-product biorefineries. J. Ind. Microbiol. Biotechnol. 35(5):367–375.

    Article  CAS  Google Scholar 

  5. Wood TM and Bhat KM (1988) Methods for measuring cellulase activities. Methods Enzymol. 160:87–117.

    Article  CAS  Google Scholar 

  6. Ghose TK (1987) Measurement of cellulase activities. Pure Appl. Chem. 59:257–268.

    Article  CAS  Google Scholar 

  7. Zhang Y-HP and Lynd LR (2006) A functionally-based model for hydrolysis of cellulose by fungal cellulase. Biotechnol. Bioeng. 94:888–898.

    Article  CAS  Google Scholar 

  8. Hong J, Ye X, and Zhang Y-HP (2007) Quantitative determination of cellulose accessibility to cellulase based on adsorption of a nonhydrolytic fusion protein containing CBM and GFP with its applications. Langmuir 23(25):12535–12540.

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  10. Coward-Kelly G, Aiello-Mazzari C, Kim S, Granda C, and Holtzapple M (2003) Suggested improvements to the standard filter paper assay used to measure cellulase activity. Biotechnol. Bioeng. 82:745–749.

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

  12. Zhang Y-HP and Lynd LR (2003) Quantification of cell and cellulase mass concentrations during anaerobic cellulose fermentation: development of an ELISA-based method with application to Clostridium thermocellum batch cultures. Anal. Chem. 75:219–227.

    Article  CAS  Google Scholar 

  13. Zhang Y-HP and Lynd LR (2005) Regulation of cellulase synthesis in batch and continuous cultures of Clostridium thermocellum. J. Bacteriol. 187:99–106.

    Article  CAS  Google Scholar 

  14. Zhang Y-HP and Lynd LR (2004) Kinetics and relative importance of phosphorolytic and hydrolytic cleavage of cellodextrins and cellobiose in cell extracts of Clostridium thermocellum. Appl. Environ. Microbiol. 70:1563–1569.

    Article  CAS  Google Scholar 

  15. Hong J, Ye X, Wang Y, and Zhang Y-HP (2008) Bioseparation of recombinant cellulose binding module-protein by affinity adsorption on an ultra-high-capacity cellulosic adsorbent. Anal. Chem. Acta 621:193–199.

    Article  CAS  Google Scholar 

  16. Zhang Y-HP and Lynd LR (2005) Determination of the number-average degree of polymerization of cellodextrins and cellulose with application to enzymatic hydrolysis. Biomacromolecules 6:1510–1515.

    Article  CAS  Google Scholar 

  17. Zhang Y-HP, Cui J-B, Lynd LR, and Kuang LR (2006) A transition from cellulose swelling to cellulose dissolution by o-phosphoric acid: evidences from enzymatic hydrolysis and supramolecular structure. Biomacromolecules 7(2):644–648.

    Article  CAS  Google Scholar 

  18. Miller GL (1959) Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal. Chem. 31:426–428.

    Article  CAS  Google Scholar 

  19. Zhang Y-HP, Schell DJ, and McMillan JD (2007) Methodological analysis for determination of enzymatic digestibility of cellulosic materials. Biotechnol. Bioeng. 96(1):188–194.

    Article  CAS  Google Scholar 

  20. Kongruang S, Han MJ, Breton CIG, and Penner MH (2004) Quantitative analysis of cellulose-reducing ends. Appl. Biochem. Biotechnol. 113–116:213–231.

    Article  Google Scholar 

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Acknowledgments

This work was made possible in part by support from the Biological Systems Engineering Department of Virginia Polytechnic Institute and State University and USDA-CSREES (2006–38909–03484) and DOE BioEnergy Science Center (BESC).

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Authors and Affiliations

  1. Biological Systems Engineering Department, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA

    Y. H. Percival Zhang, Jiong Hong & Xinhao Ye

Authors
  1. Y. H. Percival Zhang
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  2. Jiong Hong
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  3. Xinhao Ye
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Editor information

Editors and Affiliations

  1. Bioconversion Science & Technology, Oak Ridge National Laboratory, Bethel Valley Road 1, Oak Ridge, 37831-6341, U.S.A.

    Jonathan R. Mielenz

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Zhang, Y.H.P., Hong, J., Ye, X. (2009). Cellulase Assays. In: Mielenz, J. (eds) Biofuels. Methods in Molecular Biology, vol 581. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-214-8_14

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  • DOI: https://doi.org/10.1007/978-1-60761-214-8_14

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-213-1

  • Online ISBN: 978-1-60761-214-8

  • eBook Packages: Springer Protocols

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