Watkinson, Sarah C., Burton, Kerry S. and Wood, D. A. (David A.). (2001) Characteristics of intracellular peptidase and proteinase activities from the mycelium of a cord-forming wood decay fungus Serpula lacrymans. Mycological Research, Vol.10 (No.6). pp. 698-704. ISSN 0953-7562

Preview

PDF WRAP_Watkinson_Characteristics_intracellular.pdf - Requires a PDF viewer such as GSview, Xpdf or Adobe Acrobat Reader Download (377Kb)

Abstract

Serpula lacrymans is a basidiomycete cord-forming wood decay fungus which reallocates nitrogen within an extensive perennial mycelial system in response to spatial discontinuities in external nutrient supply. Intracellular stored protein is mobilised by conversion to amino acids at nutrient-poor sites within a mycelium or when a whole mycelium is starved. Intracellular peptidase and proteinase activities of the mycelium were investigated with the aim of identifying proteases specifically activated in response to a nitrogen demand. Mycelium for enzyme extraction grown as surface mats in static liquid culture was homogenised, and the extract used in assays for proteinase and peptidase with various synthetic peptide substrates conjugated to 4-nitroaniline. Activities against different substrates were characterised with respect to pH, inhibitor sensitivity, requirements for divalent metal ions, isoelectric point, and by changes in activities in starved mycelium. Four different activities were found, comprising two peptidases one of which had metalloprotease characteristics, a serine-type proteinase, and a proteinase active at pH 2.5 which was not affected by any of the inhibitors tried. Both the latter were most active in starved mycelium. Isoelectric focusing showed peaks with activities corresponding to the serine-type proteinase and one of the manganese-activated peptidases. Possible roles for these enzymes in nitrogen reallocation during mycelial foraging are discussed.

Item Type:	Journal Article
Subjects:	S Agriculture > SB Plant culture
Divisions:	Faculty of Science > Life Sciences (2010- ) > Warwick HRI (2004-2010)
Library of Congress Subject Headings (LCSH):	Serpula lacrymans, Mycelium, Peptidase, Proteinase, Nitrogen cycle, Fungi -- Hyphae, Basidiomycetes
Journal or Publication Title:	Mycological Research
Publisher:	Cambridge University Press
ISSN:	0953-7562
Date:	June 2001
Volume:	Vol.10
Number:	No.6
Page Range:	pp. 698-704
Identification Number:	10.1017/S0953756201003938
Status:	Peer Reviewed
Access rights to Published version:	Open Access
Funder:	Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC), Great Britain. Ministry of Agriculture, Fisheries and Food (MAFF)
References:	D.S. Auld, Removal and replacement of metal ion in metallopeptidases, Methods in Enzymology A. J. Barrett 248 (1995), pp. 228–242. R.J. Beynon and J.S. Bond, Proteolytic Enzymes: a practical approach, IRL Press, Oxford (1994). L. Boddy, Saprotrophic cord-forming fungi: warfare strategies and other ecological aspects, Mycological Research 97 (1993), pp. 641–655. L. Boddy and S.C. Watkinson, Wood decomposition, higher fungi, and their role in nutrient redistribution, Canadian Journal of Botany 73 (Suppl.) (1995), pp. S1377–S1383. M.M. Bradford, A rapid and sensitive method for the quantitation of microgramme quantities of protein using the principle of protein-dye binding, Analytical Biochemistry 72 (1976), pp. 248–254. K.S. Burton, M.D. Partis, D.A. Wood and C.F. Thurston, Accumulation of serine proteinase in senescent sporophores of the cultivated mushroom, Agaricus bisporus, Mycological Research 101 (1997), pp. 146–152. K.S. Burton, D.A. Wood, C.F. Thurston and P.J. Barker, Purification and characterization of a serine proteinase from senescent sporophores of the commercial mushroom Agaricus bisporus, Journal of General Microbiology 139 (1993), pp. 1379–1386. F.H. Carpenter and J.M. Vahl, Leucine aminopeptidase (bovine lens), Journal of Biological Chemistry 248 (1973), pp. 294–304. J. Dighton, Nutrient cycling by saprotrophic fungi in terrestrial habitats. In: D.T. Wicklow and B. Soderstrom, Editors, The Mycota. Vol. IV. Environmental and Microbial Relationships, Springer Verlag, Berlin (1997), pp. 271–279. G.W. Gooday, Cell walls. In: N.A.R. Gow and G.M. Gadd, Editors, The Growing Fungus, Chapman & Hall, London (1995), pp. 43–62. G.G. Khatchatourians, Biochemistry and molecular biology of entomopathogenic fungi. In: D.H. Howard and J.D. Miller, Editors, The Mycota. Vol. VI. Human and Animal Relationships, Springer Verlag, Berlin (1996), pp. 331–363. J.R. Leake and D.J. Read, Mycorrhizal fungi in terrestrial habitats. In: D.T. Wicklow and B. Soderstrom, Editors, The Mycota. Vol. IV. Environmental and Microbial Relationships, Springer Verlag, Berlin (1997), pp. 281–301. M.J. North, Comparative biochemistry of proteinases of eukaryotic organisms, Microbiological Reviews 46 (1982), pp. 308–340. J.J.R. Frausto da Silva and R.J.P. Williams, The Biological Chemistry of the Elements: the inorganic chemistry of life, Clarendon Press, Oxford (1991). C.E. Venables and S.C. Watkinson, Medium-induced changes in patterns of free and combined amino acids in mycelium of Serpula lacrymans, Mycological Research 92 (1989), pp. 273–277. C.E. Venables and S.C. Watkinson, Production and localisation of proteinases in colonies of timber decaying basidiomycete fungi, Journal of General Microbiology 135 (1989), pp. 1369–1374. R.V. Wadekar, M.J. North and S.C. Watkinson, Proteolytic activities in two wood-decaying basidiomycete fungi, Serpula lacrymans and Coriolus versicolor, Microbiology 141 (1995), pp. 1575–1583. S.C. Watkinson, Inhibition of growth and development of Serpula lacrymans by the non-metabolised amino acid analogue 2-aminoisobutyric acid, FEMS Microbiology Letters 24 (1984), pp. 247–250. S.C. Watkinson, Metabolism and differentiation in basidiomycete mycelium. In: N.A.R. Gow, Editor, The Fungal Colony, Cambridge University Press, Cambridge (1999), pp. 126–156. S.C. Watkinson, E.M. Davison and J. Bramah, The effect of nitrogen availability on growth and celluloysis by Serpula lacrymans, New Phytologist 89 (1981), pp. 295–305.
URI:	http://wrap.warwick.ac.uk/id/eprint/816

Request changes to a record

Actions (login required)

View Item