Evaluation of VOC producing Diaporthe sp. for enzyme production

As dependence on fuel increases, more people have begun searching for an alternative. Endophytes, microorganisms that live mutualistically within plants, have recently been sought after as a source of alternative fuel known as MycodieselⓇ. As the demand for alternative fuel increases, the need for non food-based feedstocks, such as recycled paper, also increases. An endophytic Diaporthe sp., isolated from Black Gum Bark and known to produce volatile organic compounds (VOCs), was screened for cellobiose, starch, and lignin degrading enzymes, which are the primary macromolecules contained in paper. Screening results for production of lignin degrading enzymes showed production of phenoloxidase using a guaiacol plate assay. Production of a starch degrading enzyme was observed using a Gram’s Iodine assay. Lastly, production of a cellobiose degrading enzyme was observed using a p-nitrophenol assay. Cellobiase was isolated from Diaporthe sp. using ammonium sulfate precipitation and centrifugation, then retested for its activity using the p-nitrophenol assay. It was concluded that the endophyte produces lignin, starch, and cellobiose degrading enzymes, with cellobiose being produced in more measurable amounts. Because cellulose is the most prominent macromolecule found in paper, future work includes isolating higher concentrations of cellulose degrading enzymes.