Research Project Summary Information
Renewable Energy Technology Options Program:Production of Lignin Peroxidase(ST8209-1)
Lignin Peroxidase (LiP) is an enzyme that has been proven to remove lignin from biomass, an important step in many biomass refining operations, including papermaking. Commercial use of this enzyme has been limited in the past because white rot fungus, which produces it, is finicky in fermentations.
Researchers hoped to develop a yeast strain, then produce LiP in the yeast, a more predictable and manageable host than white rot fungus because it is not inhibited by shear stresses commonly found in fermentation vessels, it grows more densely, and is commonly employed by commercial firms to produce other proteins and enzymes. The overall goal of this project was to develop a yeast strain that secretes LiP at yields of at least 10 mg/L. While this was not completed, significant steps along this path were accomplished.
Higher concentrations of product (1-10 g/L) in the fermentation broth of large reactors (10,000 L) will lead to considerably decreased purification costs. A successful Stage One project would have developed this strain.
Future phases would manufacture it in two L vessels at a cost of approximately $220/batch and sell it routinely through industrial channels for $1000-$2500per batch. If yields in Stage Two exceed 100 mg/L, the process would be scaled up to 1000 L.
All of the DNA cloning vectors were constructed, and the polymerase chain reaction (PCR) conditions and primers for amplification of the gene lipA were successfully employed using white-rot fungus DNA. The white-rot fungus was successfully cultured under lip-inducing conditions for the production of messenger RNA (mRNA) for reverse transcription to lipA complementary DNA (cDNA) and the array of cDNAs to be used in constructing the expression library in the yeast. Most of the remaining work involves cloning of the DNA derived from the mRNA, then screening the resulting strains for any producing appreciably higher LiP enzyme titers.
727 East Washington St. Attn: Tamara Rosanio
Syracuse, NY 13244
Indigenous/Renewable Energy Resources
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R&D - Environment & Energy Res