Skip to main content
SpringerLink
Log in

Production and initial characterisation of the xylan-degrading system of Phanerochaete chrysosporium

  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

We report the optimum conditions for the degradation of oat spelt arabinoxylan and a preliminary characterisation of the inducible xylan-degrading system of the lignin-degrading white-rot fungus Phanerochaete chrysosporium. Xylanase activity was optimal at pH 5.0 and 50°C; see attached sheet the maximum reaction velocity (Vmax) of the system was 3.86 units (U) mg−1 protein with arabinoxylan as substrate and the substrate concentration giving half Vmax (S0.5) was 0.52 mg ml−1. At concentrations of arabinoxylan greater than 15 mg ml−1 excess substrate inhibition was observed. Xylose at 0.9 mm inhibited activity to the extent of 50%. Xylanase activity increased as a function of the dilution of the enzyme preparation prior to assay. It was resolved into four peaks by using a DEAE-Biogel column; the material in these peaks differed with respect to xylan solubilisation and the formation of reducing sugars. Electrofocusing gels allowed visualisation of several bands of activity corresponding to each peak. The arabinoxylan degradation system of P. chrysosporium is therefore composed of multiple components.

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

  • Bailey M, Puls J, Poutanen K (1991) Purification and properties of two xylanases from Aspergillus oryzae. Biotechnol Appl Biochem 13:380–389

    Google Scholar 

  • Biely P (1985) Microbial xylanolytic systems. Trends Biotechnol 3:286–290

    Google Scholar 

  • Birch O (1988) Extracellular enzymes from the lignin-degrading fungus Phanerochaete chrysosporium. Ph.D. Thesis, University of Manchester

  • Blanchette RA, Abad AR, Cease KR, Farrell RL, Lourien RE, Leather TD (1990) Enzyme immunocytochemistry and ultrastructural localization of cell wall components by enzyme-gold complexes. In: Kirk TK, Chang HM (eds) Biotechnology in pulp and paper manufacture: applications and fundamental investigations. Butterworth Heinemann, Boston, pp 69–81

    Google Scholar 

  • Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:28–254

    Google Scholar 

  • Comtat J (1983) Isolation, properties and postulated role of some of the xylanases from the basidiomycete Sporotrichum dimorphosporum. Carbohydr Res 118:215–231

    Google Scholar 

  • Datta A, Battermann A, Kirk T (1991) Identification of a specific manganese peroxidase among ligninolytic enzymes secreted by Phanerochaete chrysosporium during wood decay. Appl Environ Microbiol 57:1453–1460

    Google Scholar 

  • Dobozi MS, Szakacs G, Bruschi CV (1992) Xulanase activity of Phanerochaete chrysosporium. Appl Environ Microbiol 58:3466–3471

    Google Scholar 

  • Eriksson KE (1981) Microbial degradation of cellulose and lignin. SCPI International Symposium on Wood and Pulping Chemistry (Stockholm) vol 3, pp 60–65

  • Eriksson KE, Blanchette R, Ander P (1990) Microbial and enzymatic degradation of wood and wood components. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Gascon S, Lampen J (1968) Purification of a internal invertase of yeast. J Biol Chem 243:1567–1572

    Google Scholar 

  • Gilkes NR, Henrissat B, Kilburn DG, Miller RC, Warren RAJ (1991) Domains in microbial β-1,4-glycanases: sequence conservation, function, and enzyme families. Microbiol Rev 55:303–315

    Google Scholar 

  • John M, Schmidt J (1989) Xylanases and β-xylosidase of Trichoderma lignorum. Methods Enzymol 160:662–671

    Google Scholar 

  • Johnson WC, Lindsey AJ (1939) An improved universal buffer. Analyst 64:490–492

    Google Scholar 

  • Jurasek KL, Paice MG (1988) Xylanase A of Schizophyllum commune. Methods Enzymol 160:659–662

    Google Scholar 

  • Khan AW, Tremblay D, LeDuy A (1986) Assay of xylanase and β-xylosidase activities in bacterial and fungal cultures. Enzyme Microb Technol 8:373–377

    Google Scholar 

  • Kirk TK, Farrell RL (1987) Enzymatic “combustion”: the microbial degradation of lignin. Annu Rev Microbiol 41:465–505

    Article  CAS  PubMed  Google Scholar 

  • Kluepfel D, Daigneault N, Morosoli R, Shareck F (1992) Purification and characterization of a new xylanase (xylanase C) produced by Streptomyces lividans 66. Appl Microbiol Biotechnol 36:626–631

    Google Scholar 

  • Lachke AH (1988) 1,4-β-d-Xylan xylohydrolase of Sclerotium rolfsii. Methods Enzymol 160:679–684

    Google Scholar 

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

    PubMed  Google Scholar 

  • Llanos-Palop M, Vallés S, Piñaga F, Flors A (1991) Characterization of cellulase and xylanase activities of Clostridium celerecrescens. J Chem Technol Biotechnol 51:105–114

    Google Scholar 

  • Mason JC, Birch OM, Broda P (1990) Preparation of 14C-radiolabelled lignocelluloses from spring barley of differing maturities and their solubilization by Phanerochaete chrysosporium and Streptomyces cyaneus. J Gen Microbiol 136:227–232

    Google Scholar 

  • Mishra C, Keskar S, Rao M (1984) Production and properties of extracellular endoxylanase from Neurospora crassa. Appl Environ Microbiol 48:224–228

    Google Scholar 

  • Mishra C, Forrester I, Kelley B, Burgess R, Leatham G (1990) Lentinula edodes cultures grown on a commercial solid lignocellulose substrate. Appl Microbiol Biotechnol 33:226–232

    Google Scholar 

  • Mopper R, Gindler J (1973) A new noncorrosive dye reagent for automatic sugar chromatography. Anal Biochem 56:440–442

    Google Scholar 

  • Morrisey JH (1981) Silver stain for proteins in polyacrylamide gels. A modified procedure with enhanced uniform sensitivity. Anal Biochem 117:307–310

    Google Scholar 

  • Nakayama M, Tomita Y, Susuki H, Nisizawa K (1976) Partial proteolysis of some cellulase components from Trichoderma viride and the substrate specificity of the modified products. J Biochem 79:955–966

    Google Scholar 

  • Nawaz M, Gunasekaran M (1988) Effect of peat extract on the hydrolytic enzymes of Phanerochaete chrysosporium. Resour Conserv Recycl 1:197–205

    Google Scholar 

  • Poutanen K, Puls J (1988) Characteristics of Trichoderma reesei β-xylosidase and its use in the hydrolysis of solubilized xylans. Appl Microbiol Biotechnol 28:425–432

    Google Scholar 

  • Rapp P, Wagner F (1986) Production and properties of xylan degrading enzymes from Cellulomonas uda. Appl Environ Microbiol 54:746–752

    Google Scholar 

  • Sinner M, Puls J (1978) Non-corrosive dye reagent for detection of reducing sugars in borate complex ion-exchange chromatography. J Chromatog 156:197–204

    Google Scholar 

  • Tenkanen M, Puls J, Poutanen K (1992) Two major xylanases of Trichoderma reesei. Enzyme Microb Technol 14:566–574

    Google Scholar 

  • Tuohy MG, Coughlan MP (1992) Production of thermostable xylan-degrading enzymes by Talaromyces emersonii. Bioresour Technol 39:131–137

    Google Scholar 

  • Wang P, Broda P (1992) Stable defined substrate for turbidimetric assay of endoxylanases. Appl Environ Microbiol 58:3433–3436

    Google Scholar 

Download references

Author information

Author notes

  1. José L. Copa-Patiño

    Present address: Departmento de Microbiología y Parasitología, Universidad de Alcalá de Henares, Alcalá de Henares 28871, Madrid, Spain

  2. Young G. Kim

    Present address: Department of Food Science, Miryang National Technical College, 1028 Nayi-Dong Miryang, Gyeongnam, South Korea

Authors and Affiliations

  1. Department of Biochemistry and Applied Molecular Biology, University of Manchester Institute of Science and Technology, P.O. Box 88, M60 1QD, Manchester, UK

    José L. Copa-Patiño, Young G. Kim & Paul Broda

Authors
  1. José L. Copa-Patiño
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Young G. Kim
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Paul Broda
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Correspondence to: P. Broda

Rights and permissions

Reprints and permissions

About this article

Cite this article

Copa-Patiño, J.L., Kim, Y.G. & Broda, P. Production and initial characterisation of the xylan-degrading system of Phanerochaete chrysosporium . Appl Microbiol Biotechnol 40, 69–76 (1993). https://doi.org/10.1007/BF00170431

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00170431

Keywords

Search

Navigation