Glycerol Determination in Biodiesel Fuels using Glycerol Oxidase

Glycerol, also called glycerin(e), is a byproduct or contaminant of biofuel production. Glycerol must be removed from biofuels before they can be burned in order to avoid damage to combustion engines. The standard for biodiesel is less than 0.02% by weight (ASTM D6751; ASTM International sets voluntary consensus standards for quality in many industry sectors). Standard methods for glycerol determination require skilled operators, complex sample preparation, and expensive lab equipment. Biofuel producers need facile and inexpensive methods for glycerol detection, especially in biodiesel. NECi has identified an unusual fungal enzyme, glycerol oxidase, and demonstrated that it is capable of glycerol determination in raw biodiesel at varying production stages. The Phase I project focuses on assay development and improved production/purification of the native enzyme. Prototype test kits and reagents for high-throughput labs based on the purified native enzyme will be developed in Phase I. Recombinant Glycerol Oxidase enzyme is the key target product of the Phase I/II project. Test kits based on the new enzyme will in incorporated into NECis test kit product line and marketed directly to biofuels producers. Enzyme reagent packages will be developed for autoanalyzer equipment in collaboration with the manufacturers.