New Probe Aids Enzyme Mixology for Biofuels

What’s the difference between a bartender and a biofuels researcher? A bartender uses ethanol to make cocktails, while a biofuels researcher uses cocktails to make ethanol. Researchers at the Department of Energy’s Pacific Northwest National Lab have developed a probe to help create the most efficient cocktails for biofuels makers.

Trichoderma reesei

Trichoderma reesei, an enzyme powerhouse. Credit: Department of Energy

A biofuel-making cocktail is a blend of enzymes that break down biomass (like corn stalks). And apparently the fungus Trichoderma reesei is a veritable Swiss Army knife of enzymes.  T.E., as we’ll call it, is a mesophilic soft-rot fungus which was famous in World War II as the stuff that chewed through military tents in the Pacific Theater. It contains 200 sugar molecule busting enzymes (glycoside hydrolases) including 10 that chomp cellulose and 16 that consume hemicellulose. This variety is helpful, because no single enzyme can profitably make ethanol from cellulose.

To make biofuels, companies either make or purchase custom blends of enzymes that function at the needed pH, temperature, nutrient environment, and chemical conditions. Companies like Novozymes sell optimized blends of enzymes.

But with PNNL’s probes, cocktail DIY’ers can get in on the action. Currently, enzyme assays only show the total mixture activity of all enzymes, not the activity of individual enzymes. But the activity-based probe method quickly identifies and quantifies the activity of individual enzymes in a mixture, allowing high throughput analysis with gel electrophoresis or LC-MS-based proteomics. The research showed that the different processing conditions had a significant impact on the activity of individual enzymes. Armed with this knowledge, an enzyme mixologist would be able to more quickly identify the best ingredients for their biofuels process.

Reference [free download with registration at RSC]: Lindsey N. Anderson, David E. Culley, Beth A. Hofstad, Lacie M. Chauvigné-Hines, Erika M. Zink, Samuel O. Purvine, Richard D. Smith, Stephen J. Callister, Jon M. Magnuson and Aaron T. Wright, Activity-based protein profiling of secreted cellulolytic enzyme activity dynamics in Trichoderma reesei QM6a, NG14, and RUT-C30, Molecular BioSystems, Oct. 9, 2013, DOI: 10.1039/c3mb70333a.

Author: Melody Bomgardner

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