Asian Journal of Microbiology, Biotechnology & Environmental Sciences Paper

COMPARATIVE BIOCHEMICAL AND RAPD-PCR FINGERPRINTING ANALYSIS OF LIGNINOLYTIC FUNGI PENICILL1M OXALICLIM AND PHANEROCHAETE CHRYSOSPORILIM

HARIT JHA SHAHZAD NAWAZ SYED, VILAS PARKHI, MANOJ R. BHANDARKAR AND MANDAKINI PATIL

Abstract

In present work lignin degrading activity of penicillin'', oxalicum isolate-1 and Phanerochaete chTysospornon were assessed. Comparative biochemical analysis was carried out to evaluate their utility for lignin degradation in pulp and paper industry and attempt has been made to characterize them on the basis of available PCR based molecular tools. Biochemically the two fungi differs. Comparative analysis shows that in P. chrysosporium activities of Lignin Peroxidase (42 U/mg), Manganese Peroxidase (57.2 U/mg), Xylanase (150x10-3U/mg), Cellulase (33 U/mg) and Pectinase (130 U/mg) are higher than that the activities of Manganese Peroxidase (32 U/mg), Xylanase (120x10-3) U/mg), Cellulase (19 U/mg) and l'ectinase (130 U/ mg) in P oxlicum isolate-1, however surprisingly no Lignin Peroxidase activity could be detected in the later. In the present study for the first time ligninoly tic activity is reported in P. avalicion isolate-1 hence opening a new horizon of organisms that can be exploited for commercial purpose. Considering the environmental friendly industrial importance of these ligninolytic fungi RAPD fingerprints were developed. The two fungi are different, taxonomically and phenotypically, thus as expected most of the 20 RAPD primers of OPA series showed polymorphism except primer OPA14, which amplified 1.1 kb monomorphic marker. Despite this divergence, the two fungi share common property of being lignin degraders. Since MnP is dominant enzyme in lignin degradation process, using primer sequence of known MnP of P. chrysosporium, RT-PCR analysis was carried out. 111np gene specific primer set MnP_04A amplified 1.1 kb cDNA fragments from both the organism. To correlate the results and to assign the molecular property, this candidate amplicon from P. oxalic= isolate-1 was successfully cloned and 1.1 kb fragments were obtained. Further work needs to be carried out to find out the correlation between molecular and biochemical data such as cloning and sequencing of all Milp mRNAs, sequence analysis and annotation, and to design the molecular identification tools.