Skip to main content
SpringerLink
Log in
SpringerLink
Log in

Purification and Characterization of the Total Cellulase Activities (TCA) of Cellulolytic Microorganisms

  • Protocol
  • First Online:
Cellulases

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1796))

  • 1395 Accesses

Abstract

Cellulose is the earth’s most abundant plant polysaccharide containing a large array of glucose units linked through β (1 → 4) linkages by existing in both crystalline and amorphous forms. Cellulose is widely distributed in plants, constituting up to 40–50% overall dry weight of the plant biomass. Majorly, microorganisms secrete three types of enzymes such as endoglucanases, exoglucanases, and beta-glucosidase for the hydrolysis of cellulose, contributing to the total cellulase activity. Industrially, the cellulolytic microorganisms are assessed based on their total cellulolytic activities. Similarly, total cellulase activity can also be used for the isolation and characterization of the cellulolytic microorganisms. In this chapter, we have specifically discussed about the methods used for the purification and characterization of the total cellulase activities of the microorganisms such as filter paper assay and cellulase zymogram assay. Our present chapter can be used as primer for characterizing cellulolytic abilities of cellulose-degrading microorganisms.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Lynd LR, Weimer PJ, Van Zyl WH, Pretorius IS (2002) Microbial cellulose utilization: fundamentals and biotechnology. Microbiol Mol Biol Rev 66(3):506–577

    Article  CAS  Google Scholar 

  2. Dashtban M, Maki M, Leung KT, Mao C, Qin W (2010) Cellulase activities in biomass conversion: measurement methods and comparison. Crit Rev Biotechnol 30(4):302–309

    Article  CAS  Google Scholar 

  3. Béguin P, Aubert J-P (1994) The biological degradation of cellulose. FEMS Microbiol Rev 13(1):25–58

    Article  Google Scholar 

  4. Horn SJ, Vaaje-Kolstad G, Westereng B, Eijsink V (2012) Novel enzymes for the degradation of cellulose. Biotechnol Biofuels 5(1):1

    Article  Google Scholar 

  5. Baldrian P, Valášková V (2008) Degradation of cellulose by basidiomycetous fungi. FEMS Microbiol Rev 32(3):501–521

    Article  CAS  Google Scholar 

  6. Bégum P, Lemaire M (1996) The cellulosome: an exocellular, multiprotein complex specialized in cellulose degradation. Crit Rev Biochem Mol Biol 31(3):201–236

    Article  Google Scholar 

  7. Liao JC, Mi L, Pontrelli S, Luo S (2016) Fuelling the future: microbial engineering for the production of sustainable biofuels. Nat Rev Microbiol 14(5):288–304

    Article  CAS  Google Scholar 

  8. Waterborg JH (2009) The Lowry method for protein quantitation. In: Walker JM (ed) The protein protocols handbook. Human Press, New York, pp 7–10

    Chapter  Google Scholar 

  9. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  10. Zhang YP, Hong J, Ye X (2009) Cellulase assays. Methods Mol Biol 581:213–231

    Article  CAS  Google Scholar 

  11. Xiao Z, Storms R, Tsang A (2004) Microplate-based filter paper assay to measure total cellulase activity. Biotechnol Bioeng 88(7):832–837

    Article  CAS  Google Scholar 

  12. Bakare M, Adewale I, Ajayi A, Shonukan O (2005) Purification and characterization of cellulase from the wild-type and two improved mutants of Pseudomonas fluorescens. Afr J Biotechnol 4(9)

    Google Scholar 

  13. Gaur R, Tiwari S (2015) Isolation, production, purification and characterization of an organic-solvent-thermostable alkalophilic cellulase from Bacillus vallismortis RG-07. BMC Biotechnol 15(1):19

    Article  Google Scholar 

  14. Du R, Su R, Zhang M, Qi W, He Z (2014) Cellulase recycling after high-solids simultaneous saccharification and fermentation of combined pretreated corncob. Front Energy Res 2:24

    Article  Google Scholar 

  15. Ang SK, Shaza E, Adibah Y, Suraini A, Madihah M (2013) Production of cellulases and xylanase by Aspergillus fumigatus SK1 using untreated oil palm trunk through solid state fermentation. Process Biochem 48(9):1293–1302

    Article  CAS  Google Scholar 

  16. Geib SM, Tien M, Hoover K (2010) Identification of proteins involved in lignocellulose degradation using in gel zymogram analysis combined with mass spectroscopy-based peptide analysis of gut proteins from larval Asian longhorned beetles, Anoplophora glabripennis. Insect Sci 17(3):253–264

    Article  CAS  Google Scholar 

  17. Thomson I (2015) Chamaiporn Champasri, Thongchai Champasri and Khanutsanan Woranam. Asian J Biochem 10(5):190–204

    Article  Google Scholar 

  18. Anuradha Jabasingh S, Varma S, Garre P (2014) Production and purification of cellulase from Aspergillus nidulans AJSU04 under solid-state fermentation using coir pith. Chem Biochem Eng Q 28(1):143–151

    Google Scholar 

  19. Gavrilov SN, Stracke C, Jensen K, Menzel P, Kallnik V, Slesarev A, Sokolova T, Zayulina K, Bräsen C, Bonch-Osmolovskaya EA (2016) Isolation and characterization of the first xylanolytic hyperthermophilic euryarchaeon Thermococcus sp. strain 2319x1 and its unusual multidomain glycosidase. Front Microbiol 7:552

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biology, Lakehead University, Thunder Bay, ON, Canada

    Ayyappa Kumar Sista Kameshwar & Wensheng Qin

Authors
  1. Ayyappa Kumar Sista Kameshwar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Wensheng Qin
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Wensheng Qin .

Editor information

Editors and Affiliations

  1. Section of Sustainable Biotechnology, Department of Chemistry and Bioscience, Aalborg University, Copenhagen, Denmark

    Mette Lübeck

Rights and permissions

Reprints and permissions

Copyright information

© 2018 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Kameshwar, A.K.S., Qin, W. (2018). Purification and Characterization of the Total Cellulase Activities (TCA) of Cellulolytic Microorganisms. In: Lübeck, M. (eds) Cellulases. Methods in Molecular Biology, vol 1796. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7877-9_18

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-7877-9_18

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7876-2

  • Online ISBN: 978-1-4939-7877-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation