SBIR-STTR Award

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.

Whenever biodiesel is made, ten percent of the reaction is the by-product glycerol (also called glycerin). Glycerol has many uses, but it is not a fuel: burning glycerol in any engine will cause irreversible damage. So glycerol must be removed from biodiesel before it can be sold or used. ASTM Intermational, the trade association that sets standards for purity of many industrial products, has set the limit of glycerol in biodiesel at 0.02 percent. There is no wuick and easy way to measure the glycerol content of biodiesel. NECi has identified an enzyme that can do the job. In order to make the test practical, commercial quantities of enzyme of consistently reliable quality is needed. The goal of this project is to develop protein expression processes for making pure and stable enzyme. At the same time, we will develop reagents and simple test kits. Reagents are targeted to biodiesel process development groups and producers. Simple kits for use on site are aimed at small scale producers such as on-farm sites, and fuel testers. The goals will help producers design better fuel production processes, help regulatory personnel monitor fuel QC, and prevent engine damage from partially purified biodiesel. The overall goal is to help make biodiesel a more practical, commercially competitive fuel choice.