IN VITRO SCREENING OF CELL WALL DEGRADING ENZYME PRODUCTIVITY FROM FUNGAL CULTURE FILTRATES ON DEPROTEINISED PLANT FLUID BY CUP PLATE ASSAY

Abstract

    The present  study  purpose  is  to  evaluate  the  potentiality  of  the  Deproteinised  juice made  from  selected  plants to  induce  the  enzyme productivity  by growing  the   fungi  on  it. It  is  because, in  earlier  findings,  the   DPJ  was  found potential in  enhancing  fungi  and  the  plant  growth  when  used as  the  medium. The internal  factors  are  responsible in  DPJ  which induces  plants  and  fungi  growth.  During  the  process of  Green  Crop  Fractionation (GCF), the deproteinised (DPJ) obtained from  tissues of cabbage, beet, lucerne (Alfalfa), carrot and Anathum (Dill) forages left after leaf protein extraction employed as a medium for the cultivation of mycelial biomass of  Penicillium and Aspergillus fungi. The  mycelial  growth in vitro was  compared with the glucose nitrate medium. The culture filtrates were used to screen different secreted hydrolytic or  cell wall degrading  enzymes.. The agar ‘cup‐plate’ diffusion technique has been applied to the quantitative determination of enzyme activity, principally to amylase, cellulase and protease. With all enzymes so far examined, the relationship between diameter of zone over a wide range secreted quantitatively was  examined.  All fungi grew well on DPJ in comparison to their growth on glucose nitrate (GN) medium. Comparatively with  GN  medium, lucerne DPJ  was found having more mycelial cellular  proliferation.  When the fungi grown on different concentrations of substrates, enriched  with  carboxymethyl cellulose, casein and starch in deproteinised leaf extracts and GN medium, it  was found  that  there  was the enhancement in the mycelial dry weight  grown on  DPJ as compared with glucose nitrate medium. Penicillium showed more  yield of enzyme activities especially of  cellulases and amylases as compared to protease  by  cup  plate  method. There  was  enhancement of  mycelial  biomass when  the  concentrations  of  substrates increased  from  1%  to  2%,  while  there was no  change  in  the  activity  of  all enzymes  by  increasing  the  concentrations of  substrates. Activities  of the enzymes in vitro can be indicative of the pattern of organogenesis in callus cultures in  further  studies by  fungal  culture  filtrates  cultured  on   deproteinised fluid  medium.