Skip to main content
SpringerLink
Log in

Purification and characterization of a high molecular weight endoxylanase from the solid-state culture of an alkali-tolerant Aspergillus fumigatus MKU1

  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Summary

A high molecular weight endoxylanase (XylF2) from the solid state culture of Aspergillus fumigatus MKU1 was purified to homogeneity by a combination of tube gel electrophoresis and electroelution methods. The purity was demonstrated by SDS-PAGE and the molecular mass of the XylF2 was found to be 66 kDa. The optimal pH and temperature for activity were 5.0 and 90 °C, respectively. The apparent K m and V max values of XylF2 with oat spelt xylan as substrate were 1.6 mg/ml and 3.25 mmol/min/mg protein respectively. The enzyme showed high activity towards oat spelt xylan while negligible activity was observed on carboxymethylcellulose. The activity of XylF2 was strongly inhibited by Hg2+, Ni2+, Zn2+, SDS and N-bromosuccinimide and stimulated by l-cysteine and iodoacetamide. The hydrolysis of oat spelt xylan by XylF2 released only xylo-oligosaccharides.

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

Similar content being viewed by others

References

  • Anthony T., Chandra Raj K., Rajendran A., Gunasekaran P., 2003 High molecular weight cellulase-free xylanase from alkali-tolerant Aspergillus fumigatus AR1 Enzyme and Microbial Technology 32:647–654

    Article  CAS  Google Scholar 

  • Bailey M.J., Biely P., Poutanen K., 1992 Interlaboratory testing of methods for asssay of xylanase activity Journal of Biotechnology 23:257–270

    Article  CAS  Google Scholar 

  • Baraznenok V.A., Becker E.G., Ankudimova N.V., Okunev N.N., 1999 Characterization of neutral xylanases from Chaetomium cellulolyticum and their biobleaching effect on eucalyptus pulp Enzyme and Microbial Technology 25:651–659

    Article  CAS  Google Scholar 

  • Belancic A., Scarpa J., Peirano A., D´ı ıaz R., Steiner J., Eyzaguirre J., 1995 Penicillium purpurogenum produces several xylanases: Purification and properties of two of the enzymes Journal of Biotechnology 41:71–79

    Article  CAS  Google Scholar 

  • Biely P., 1985 Microbial xylanolytic systems Trends in Biotechnology 247:286–290

    Article  Google Scholar 

  • Cardoso O.A.V., Filho E.X.F., 2003 Purification and characterization of a novel cellulase-free xylanase from Acrophialophora nainiana FEMS Microbiology Letters 223:309–314

    Article  CAS  Google Scholar 

  • Carmona E.C., Brochetto-Braga M.R., Pizzirani-Kleiner A.A., Jorge J.A., 1998 Purification and biochemical characterization of an endoxylanase from Aspergillus versicolor FEMS Microbiology Letters 166:311–315

    Article  CAS  Google Scholar 

  • Deshpande V., Hinge J., Ray M., 1990 Chemical modification of xylanases: evidence for essential tryptophan and Cyesteine residues at the active site Biochimica et Biophysica Acta 1031:172–177

    Google Scholar 

  • Elegir G., Szakacs G., Jeffries T.W., 1994 Purification, characterization and substrate specificities of multiple xylanases from Streptomyces sp. Strain B-12-2 Applied and Environmental Microbiology 60:2609–2615

    CAS  Google Scholar 

  • Filho E.X.F., Puls, J. & Coughlan, M.P. 1993 Biochemical characteristics of two endo-β-1,4-xylanasis produced by Penicillium capsulatum. Journal of Industrial Microbiology 11, 171–180

    Google Scholar 

  • Fournier A.R., Frederick M.M., Frederick J.R., Reilly P.J., 1985 Purification and characterization of endoxylanases from Aspergillus niger. Part III. An enzyme of pI 3.65 Biotechnology and Bioengineering 27:539–545

    Article  Google Scholar 

  • Gilbert H.J., Hazlewood G.P., 1993 Bacterial cellulases and xylanases Journal of General Microbiology 139:187–194

    CAS  Google Scholar 

  • Grabski A.C., Jeffries T.W., 1991 Production, purification and characterization of β-(1–4)-endoxylanase of Streptomyces roseiscleroticus Applied and Environmental Microbiology 57:987–992

    CAS  Google Scholar 

  • Khasin A., Alchanati I., Sholam Y., 1993 Purification and characterization of a thermostable xylanase from Bacillus stearothermophilus T-6 Applied and Environmental Microbiology 59:1725–1730

    CAS  Google Scholar 

  • Kulkarni N., Shendye A., Rao M., 1998 Molecular and biotechnological aspects of xylanases FEMS Microbiology Reviews 23:411–456

    Article  Google Scholar 

  • Kulkarni N., Vaidya A., Rao M., 1999 Extractive cultivation of recombinant Escherichia coli using aqueous two phase systems for production and separation of extracellular xylanase Biochemical and Biophysical Research Communications 255:274–278

    Article  CAS  Google Scholar 

  • Laemmli U.K., 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Nature 227:680–685

    Article  CAS  Google Scholar 

  • Lenartovicz V., de Souza C.G.M., Moreira F.G., Peralta R.M., 2002 Temperature effect in the production of multiple xylanases by Aspergillus fumigatus Journal of Basic Microbiology 42:388–395

    Article  CAS  Google Scholar 

  • Lin J., Ndlovu L.M., Singh S., Pillay B., 1999 Purification and biochemical characteristics of β-D-xylanase from a thermophilic fungus Thermomyces lanuginosus-SSBP Biotechnology and Applied Biochemistry 30:73–79

    CAS  Google Scholar 

  • Maheshwari R., Bharadwaj G., Bhat M.K., 2000 Thermophilic fungi: Their physiology and enzymes Microbiology and Molecular Biology Reviews 64:461–488

    Article  CAS  Google Scholar 

  • Miller G.L., 1959 Use of dinitrosalicylic acid reagent for the determination of reducing sugar Analytical Chemistry 31:426–428

    Article  CAS  Google Scholar 

  • Moreau A., Shareck F., Kluepfel D., Morosoli R., 1994 Increase in catalytic activity and thermostability of the xylanase A of Streptomyces lividans 1326 by site-specific mutagenesis Enzyme and Microbial Technology 16:420–424

    Article  CAS  Google Scholar 

  • Royer J.C., Nakas J.P., 1989 Xylanase production by Trichoderma longibrachiatum Enzyme and Microbial Technology 11:405–410

    Article  CAS  Google Scholar 

  • Ruiz M.C., Di Pietro A., Roncero M.I.G., 1997 Purification and characterization of an acidic endo-β-1,4-xylanase from the tomato vascular pathogen Fusarium oxysporum f. sp. lycopersici FEMS Microbiology Letters 148:75–82

    Article  CAS  Google Scholar 

  • Ryan S.E., Nolan K., Thompson R., Gubitz G.M., Savage A.V., Tuohy M.G., 2003 Purification and characterization of a new low molecular weight endoxylanase from Penicillium capsulatum Enzyme and Microbial Technology 33:775–785

    Article  CAS  Google Scholar 

  • Saha B.C., 2001 Xylanase from a newly isolated Fusarium verticillioides capable of utilizing corn fiber xylan Applied Microbiology and Biotechnology 56:762–766

    Article  CAS  Google Scholar 

  • Salles B.C., Cunha R.B., Fontes W., Sousa M.V., Filho E.X.F., 2000 Purification and characterization of a new xylanase from Acrophialophora nainiana Journal of Biotechnology 81:199–204

    Article  CAS  Google Scholar 

  • Sa-Pereira P., Duarte J., Costa-Ferreira M., 2000 Electroelution as a simple and fast protein purification method: isolation of an extracellular xylanase from Bacillus sp. CCMI 966 Enzyme and Microbial Technology 27:95–99

    Article  CAS  Google Scholar 

  • Senthilkumar S.R., Ashokumar B., Chandra Raj K., Gunasekaran P., 2004 Purification and characterization of a low molecular weight endoxylanase from solid state cultures of alkali-tolerant Aspergillus fischeri Biotechnology Letters 26:1283–1287

    Article  CAS  Google Scholar 

  • Shei J.C., Fratzke A.R., Frederick M.M., Frederick J.R., Reilly P.J., 1985 Purification and characterization of endoxylanases from Aspergillus niger. Part II. An enzyme of pI 4.5 Biotechnology and Bioengineering 27:533–538

    Article  CAS  Google Scholar 

  • Silva C.H.C., Puls J., de Sousa V.M., Filho E.X.F., 1999 Purification and characterization of a low molecular weight xylanase from solid-state cultures of Aspergillus fumigatus fresenius Revista de Microbiologia 30:114–119

    Google Scholar 

  • Silveira F.Q.P., Sousa M.V., Ricart C.A., Milagres A.M.F., Medeiros C.L., Filho E.X.F., 1999 A new xylanase from a Trichoderma harzianum strain Journal of Industrial Microbiology and Biotechnology 23:682–685

    Article  CAS  Google Scholar 

  • Thiagarajan, S., Jeya, M. & Gunasekaran, P. 2005 Purification and characterization of an endoxylanase from the solidstate culture of an alkalitolerant Aspergillus fumigatus MKU1 Indian Journal of Biotechnology: in press

  • Tuohy M.G., Laffey C.D., Coughlan M.P., 1994 Characterization of the individual components of the xylanolytic enzyme system of Talaromyces emersonii Bioresource Technology 50:37–42

    Article  CAS  Google Scholar 

  • Viikari L., Kantelinen A., Sundquist J., Linko M., 1994 Xylanases in bleaching: from an idea to the industry FEMS Microbiology Reviews 13:335–350

    Article  CAS  Google Scholar 

  • Wong K.K.Y., Tan L.U.L., Saddler J.N., 1988 Multiplicity of β-1,4-xylanases in microorganisms: functions and applications Microbiological Reviews 52:305–317

    CAS  Google Scholar 

Download references

Acknowledgement

Authors thank Department of Science and Technology, New Delhi, for the financial support through the project (No: SP/SO/A-20/2000).

Author information

Authors and Affiliations

  1. Department of Genetics, School of Biological Sciences, Madurai Kamaraj University, 625021, Madurai, India

    S. Thiagarajan, M. Jeya & P. Gunasekaran

Authors
  1. S. Thiagarajan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Jeya
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. P. Gunasekaran
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to P. Gunasekaran.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Thiagarajan, S., Jeya, M. & Gunasekaran, P. Purification and characterization of a high molecular weight endoxylanase from the solid-state culture of an alkali-tolerant Aspergillus fumigatus MKU1. World J Microbiol Biotechnol 22, 487–492 (2006). https://doi.org/10.1007/s11274-005-9061-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-005-9061-9

Keywords

Search

Navigation