Performing Department
(N/A)
Non Technical Summary
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.
Animal Health Component
(N/A)
Research Effort Categories
Basic
20%
Applied
50%
Developmental
30%
Goals / Objectives
Glycerol, also called glycerin(e), is a byproduct or contaminant of biofuel production. Glycerol must be removed from biofuels before they can 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; e.g. ASTM D6584-09, glycerol by gas chromatography. Several of NECis current customers are engaged in some aspect of biofuel production, from catalyst development to process monitoring to systems for growth of marine algae. These customers 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 will focus on assay development and improved production/purification of native enzyme. 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.
Project Methods
The project is an application of advanced protein purification methods for production of an enzyme, Glycerol Oxidase, to be used as a tool for analytical chemistry. Glycerol Oxidase (GlyOx) is a complex protein that has not been fully studied in the technical literature. GlyOx is rare, and difficult to purify in stable form. Advanced techniques in biochemistry and protein production and characterization are the initial methods of interest in the initial stages of the project. As material is produced, development of analytical methods using the purified protein will come into play. The GlyOx-based glycerol assay will be tested in-house on biofuel process samples obtained from biofuel processors and other potential customers. Biochemistry and analytical chemistry methods are keys to the Phase I project. Molecular biology and other biotechnology techniques (e.g. cloning and expression of GlyOx) will be used in Phase II.