Abstract
The chitinase andN-acetylglucosaminidase activities in cell-wall-bound and free fractions in the dimorphic fungusBenjaminiella poitrasii were studied as a function of morphological (unicellular yeast-mycelium) transition. The specific activities of chitinases of cell-wall-free, particularly in the membrane fraction, were significantly different in the yeast and mycelial forms. During the yeast-mycelium transition, theN-acetylglucosaminidase activity isolated in a membrane preparation increased steadily. The activity of the yeast cells (0.83±0.17 nkat/mg protein) increased 17-fold to 14.2±1.7 nkat/mg protein in 1-d-old mycelial cells. The endochitinase activity increased 12-fold between 6 and 12 h and thereafter practically remained unchanged up to 24 h. A reverse trend in the chitinolytic activities was observed during the mycelium-yeast transition. Isoelectrofocussing (pH range 3.5–10) of mixed membrane fraction free of particulate fraction of parent and morphological (y-5, yeast-form) mutant cells separated endochitinase andN-acetylglucosaminidase activity into two pH ranges,viz. 4.3–5.7 and 6.1–7.7, respectively. The predominantN-acetylglucosaminidase activity observed at pH 6.9 and 7.1 for the parent strain membrane fraction was undetected in the mutant preparation. The results suggested that the membrane-bound (either tightly or loosely) chitinolytic enzymes, particularly,N-acetylglucosaminidase, significantly contributed to the morphological changes inB. poitrasii.
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Adams D.J., Causier B.E., Mellor K.J., Keer V., Milling R., Dada J.: Regulation of chitin synthase and chitinase in fungi. pp. 15–25 in R.A.A. Muzzarelli (Ed.):Chitin Enzymology. European Chitin Society, Ancona 1993.
Balasubramanian R., Manocha M.S.: Cytosolic and membrane-bound chitinases of two mucoraceous fungi: a comparative study.Can. J. Microbiol.132, 331–338 (1992).
Binks P.R., Robson G.D., Goosey M.W., Humphreys A.M., Trinci A.P.J.: Chitin synthesis inFusarium graminearum and its inhibition by edifenphos.J. Gen. Microbiol.137, 615–620 (1990).
Deshpande M.V.: Proteinases in fungal morphogenesis.World J. Microbiol. Biotechnol.8, 242–250 (1992).
Deshpande M.V., O'Donnell R., Gooday G.W.: Regulation of chitin synthase activity in the dimorphic fungusBenjaminiella poitrasii by external osmotic pressure.FEMS Microbiol. Lett.152, 327–332 (1997).
Gooday G.W., Zhu W.-Y., O'Donnell R.W.: What are the roles of chitinases in the growing fungus?FEMS Microbiol. Lett.100, 387–392 (1992).
Humphreys A.M., Gooday G.W.: Properties of chitinase activities fromMucor mucedo: evidence for a membrane-bound zymogenic form.J. Gen. Microbiol.130, 1359–1366 (1984a).
Humphreys A.M., Gooday G.W.: Phospholipid requirement of microsomal chitinase fromMucor mucedo.Curr. Microbiol.11, 187–190 (1984b).
Jackson D.J., Saunders V.A., Gooday G.W., Humphreys A.M.: Chitinase activities from yeast and hyphal cells ofCandida albicans.Mycol. Res.100, 321–327 (1996).
Khale A., Deshpande M.V.: Dimorphism inBenjaminiella poitrasii: cell wall chemistry of parent and two stable yeast mutants.Antonie van Leeuwenhoek62, 299–307 (1992).
Khale A., Srinivasan M.C., Deshpande M.V.: Significance of NADP/NAD-glutamate dehydrogenase ratio in the dimorphic behavior ofBenjaminiella poitrasii and its morphological mutants.J. Bacteriol.174, 3723–3728 (1992).
Khale A., Srinivasan M.C., Deshmukh S.S., Deshpande M.V.: Dimorphism inBenjaminiella poitrasii: isolation and characterization of morphological mutants.Antonie van Leeuwenhoek57, 37–41 (1990).
Kopeček P., Raclavský V.: Comparison of chitin content in the apical and distal parts of fungal hyphhae inBasidiobolus ranarum, Neurospora crassa andCoprinus sterquilinus.Folia Microbiol.44, 397–400 (1999).
Kuranda M.J., Robbins P.W.: Chitinase is required for cell separation during growth ofSaccharomyces cerevisiae.J. Biol. Chem.266. 19758–19767 (1991).
Lima L.H.C., De Marco J.L., Ulhoa C.J., Felix C.R.: Synthesis of aTrichoderma chitinase which affects theSclerotium rolfsii andRhizoctonia solani cell walls.Folia Microbiol.44, 45–49 (1999).
O'Donnell R.W.: Chitinolytic enzymes ofCandida albicans. PhD Thesis. University of Aberdeen UK (1991).
Pedraza-Reyes M., Lopez-Romero E.: Purification and some properties of two forms of chitinase from mycelial cells ofMucor rouxii.J. Gen. Microbiol.135, 211–218 (1989).
Pedraza-Reyes M., Lopez-Romero E.: Detection of nine chitinase species in germinating cells ofMucor rouxii.Curr. Microbiol.22, 43–46 (1991).
Rast D.M., Horsch M., Furter R., Gooday G.W.: Complex chitinolytic system in exponentially growing mycelium ofMucor rouxii: properties and function.J. Gen. Microbiol.137, 2797–2810 (1991).
Reese E.T., Maguire A.: Increase in cellulase yields by addition of surfactants to cellobiose cultures ofTrichoderma viride.Develop. Ind. Microbiol.12, 212–224 (1971).
Reissig J.I., Strominger J.L., Leloir L.F.: A modified colorimetric method for the estimation ofN-acetylamino sugars.J. Biol. Chem.217, 959–966 (1955).
Sahai A.S., Manocha M.S.: Chitinases of fungi and plants: their involvement in morphogenesis and host-parasite interaction.FEMS Microbiol. Rev.11, 317–338 (1993).
Sathivel C., Lachke A., Radhakrishnan S.: Purification of α-arabinofuranosidase using a single-column mini-scale isoelectric focusing unit.J. Chromatogr.705, 400–402 (1995).
Sullivan P.A., McHigh N.J., Romana L.K., Shepherd M.G.: The secretion ofN-acetylglucosaminidase during germ tube formation inCandida albicans.J. Gen. Microbiol.130, 2213–2218 (1984).
Vyas P.R., Deshpande M.V.: Chitinase production byMyrothecium verrucaria and its significance for fungal mycelia degradation.J. Gen. Appl. Microbiol.35, 343–350 (1989).
Yanai K., Kakaya N., Kojima N., Horiuchi H., Ohta A., Takagi M.: Purification of two chitinases fromRhizopus oligosporus and isolation and sequencing of the encoding genes.J. Bacteriol.174 7398–7406 (1992).
Žnidaršič P., Marošek N., Pavko A.: Chitin contents in different morphological forms ofRhizopus nigricans.Folia Microbiol.44, 557–560 (1999).
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Ghormade, V.S., Lachke, S.A. & Deshpande, M.V. Dimorphism inBenjaminiella poitrasii: Involvement of intracellular endochitinase andN-acetylglucosaminidase activities in the yeast-mycelium transition. Folia Microbiol 45, 231–238 (2000). https://doi.org/10.1007/BF02908950
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DOI: https://doi.org/10.1007/BF02908950