Effect of cuticular and non cuticular substrates on the protease production in submerged cultures of high and low virulent isolates of Metarhizium anisopliae over ten days growth and autolysis phases
Abstract
Protease production pattern, the key factor governing virulence of entomopathogenic fungi was studied in the more virulent and less virulent isolates of the entomopathogenic fungus Metarhizium anisopliae using cuticles of Cockroach, Prawn, Casein or Glucose as substrates. Protease production, efficiency of growth and degree of autolysis were recorded during growth and autolysis phases. The noncuticular substrate casein demonstrated higher value of protease when compared to the samples with cuticular substrates of cockroach prawn. Effective utilization of exogenous carbon source from casein, glucose and cockroach supplements and the consequent increase in proteolytic activity detected in the high virulent isolate may be due to induction of the virulence related factors by the supplements. On the other hand, the low virulent isolate M10 showed marginal increase in proteolytic activity.
Keywords
Full Text:
PDFReferences
Charnley A K (1984) Physiological aspects of destructive pathogens in insects by fungi: a speculative review. In: Anderson (JM), Rayner (ADM) and Walton DWH (eds). Invertebrate microbial interactions No. 6: British mycological society symposium Cambridge. Cambridge University press, pp. 229-270
St Leger RJ, Charnley AK and Cooper RM (1987) Characterization of cuticle degrading proteases produced by the entomopathogen, Metarhizium anisopliae. Arch Biochem Biophys 253:221-232
Goettel MS, St Leger RJ, Rizzo NW, Staples RC and Roberts DW (1989) Ultrastructural localization of a cuticle degrading protease produced by the entomopathogenic fungus M. anisopliae during penetration of host (Manduca sexta) cuticle. J Gen Microbiol 135:2233-2239
Qazi SS and Khachatourians GG (2007) Hydrated conidia of Metarhizium anisopliae releases a family of metalloproteases. J Invertebr Pathol 95(1): 48-51
Oh YS, Shih IL, Tzeng YM and Wang SL (2000) Protease produced by Pseudomonas aeruginosa K-187 and its application in the deproteinization of shrimp and crab shell wastes. Enzyme Microb Tech 27:3-10
Rao KY, Lub SC, Li BL and Tzeng YM (2006) Enhanced production of an extracellular protease from Beauveria bassiana by optimization of cultivation processes. Biochem Eng J 28:57-66
Pinto FG da S, Fungaro MHP, Ferreira VIMC and Furlaneto MC (2002) Genetic variation in the cuticle-degrading protease activity of the entomopathogen Metarhizium flavoviride. Genet Mol Biol 25(2):231-234
Gillespie JP, Bateman R and Charnley AK (1998) Role of cuticle degrading proteases in the virulence of Metarhizium spp. for the desert locust, Schistocerca gregaria. J Invertebr Pathol 71:128-137
Campos RA, Arruda W, Boldo JT, de Silva MV, de Barros NM, de Azevedo JL, Schrank A and Vainstein MH (2005) Boophilus microplus Infection by Beauveria amorpha and Beauveria bassiana: SEM Analysis and Regulation of Subtilisin-like Proteases and Chitinases. Curr Microbiol 50:257-261
Shah FA and Butt TM (2005) Influence of nutrition on the production and physiology of sectors produced by the insect pathogenic fungus Metarhizium anisopliae. FEMS Microbiol Lett 250:201–207
G.V.S. Bhanu Prakash, Studies on the entomopathogenic fungus Metarhizium anisopliae (Metchnikoff) for virulence against the two lepidopteran pests – Biochemical characterization and expression of selected genes during pathogenesis. Ph. D. thesis Submitted to Andhra University (2008).
Söderhäll K and Unestam T (1975) Properties of extracellular enzymes form Ampharomyces astaci and their relevance in the penetration process of crayfish cuticle. Physiol Plantarum 35:140-146
Nelson N (1944) A photometric adaptation of the Somogy method for the determination of glucose. J Biol Chem 153:375-380
Clarkson JM and Charnley AK (1996) New insights into the mechanisms of fungal pathogenesis in insects. Trends Microbiol 4:197-203
El-Sayed GN, Ignoffo CM, Leathers TD and Gupta SC (1993) Cuticular and non cuticular substrate influence on expression of cuticle degrading enzymes form conidia of an entomopathogenic fungus, Nomuraea releyi. Mycopathol 122:79-87
Paterson IC, Charnley AK, Cooper RM and Clarkson JM (1994) Specific induction of a cuticle-degrading protease of the insect pathogenic fungus Metarhizium anisopliae. Microbiol 140:185-189
St Leger RJ, Bidochka MJ and Roberts DW (1994) Isoforms of the cuticle degrading Pr1 proteinase and production of a metalloproteinase by Metarhizium anisopliae. Arch Biochem Biophys 313:1-7
Magalhães BP, Faria M, Tigano MS and Sobral BWS (1997) Characterization and virulence of a Brazilian isolate of Metarhizium flavoviride Gams and Rozsypal (Hyphomycetes). Members of Entomological Society Canada 171:313-321
St Leger RJ, Joshim L and Roberts DW (1998) Ambience pH is a major determinant in the expression of cuticle degrading enzymes and hydrophobin by M. anisopliae. Appl Environ Microb 64:709-713
Moreira KA, Cavalcanti MTH, Duarte HS, Tambourgi EB, de Melo EHM, Silva VL, Porto ALF and de Lima Filho JL (2001) Partial characterization of proteases from Streptomyces clavuligerus using an inexpensive medium. Braz J Microbiol 32:215-220
St Leger RJ, Nelson JO and Screen SE (1999) The entomopathogenic fungus Metarhizium anisopliae alters ambient pH, allowing extracellular protease production and activity. Microbiol 145:2691-2699
Copyright (c) 2014 Annals of Plant Sciences

This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

