Production and Purification of Lignin Peroxidase from Bacillus Megaterium and Its Application in Bioremidation

Lignin is the second most abundant renewable biopolymer in nature after cellulose, yet it is degraded by only a small number of microorganisms. Ligninolytic enzymes are involved in the degradation of the complex and recalcitrant polymer lignin. This group of enzymes are highly versatile in nature and they find application in a wide variety of industries. In the present study lignin peroxidase producing bacteria Bacillus megaterium has been isolated and identified based on their morphological and biochemical tests. Further, Optimization of physical parameters like pH (7), temperature (37 o C), inoculum size (6%), agitation speed (180rpm) and nutritional parameters like carbon (lignin-1.5%) and nitrogen (peptone-2%) was carried out. Lignin Peroxidase was purified by using ultra filtration, ammonium sulphate precipitation and dialysis. The purity was checked by SDS-PAGE and the molecular weight of the enzyme was 65kDa. Bacillus megaterium decolorized the dyes cango red, methylene blue and malachite green to 94.90%, 63.23% and 6.40% within 96hrs of incubation. In waste water treatment Bacillus megaterium effectively acted on paper & pulp, textile red and textile blue samples with decolouration percentage of 93.70%, 48.57% and 31.64% respectively within 96hrs of incubation.