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PNNL team devises probe enabling rapid design of enzyme cocktails for maximum biomass deconstruction for biofuels

A team at Pacific Northwest National Laboratory (PNNL) has devised an activity-based probe that can rapidly identify optimal conditions for the maximum enzymatic deconstruction of lignocellulose. The probe approach promises to facilitate the rapid production of enzyme cocktails for high-efficiency lignocellulose deconstruction to support high-yield biofuel production, the researchers report in a paper in the RSC journal Molecular BioSystems.

The findings open the possibility that laboratory research that now takes months could be reduced to days, and that scientists will be able to assess more options for biofuel development than is possible today.

Many of today’s efforts revolve around the fungus Trichoderma reesei, a focus of biofuel development because of its ability to produce enzymes that chew through molecules such as complex sugars.

The breakdown of large sugar polymers into smaller compounds that can then be further converted to fuel compounds is the final, crucial step in the effort to make fuels from materials like switchgrass and corn stalks.

The ultimate goal is to begin with a plant material like corn stalks, for instance, and to subject it to a cocktail of enzymes that would convert those plants to fuel. It takes a series of steps to do that, and the cost has to come down if these fuels are to compete seriously with traditional hydrocarbon-based fuels.

—chemist Aaron Wright, PNNL team leader

T. reesei makes dozens of enzymes, each of which attacks the lignocellulosic structure differently. Chemists are trying to combine and improve upon the best ones to create a potent enzyme cocktail that accomplishes the task extremely efficiently, thus reducing the cost of producing biofuels.

Wright’s study focused on a subset of the fungus’ enzymes known as glycoside hydrolases. These enzymes break down complex sugars into simple sugars, a key step in the fuel production process.

To assess the effectiveness of mixtures of these enzymes, scientists must either measure the overall performance of the mixture, or they must test the component enzymes one at a time to see how each reacts to different conditions such as temperature, pressure and pH.

Wright’s team developed a way to measure the activity of each of the ingredients simultaneously, as well as the mixture overall. Instead of needing to run a series of experiments, each focusing on a separate enzyme, the team runs one experiment and tracks precisely how each of dozens of enzymes reacts to changing conditions.

A series of experiments detailing the activity of 30 enzymes, for example, now might be accomplished in a day or two with the new technology, compared to several months using today’s commonplace methods, the scientists said.

The core of the system, known as activity-based protein profiling, is a chemical probe that binds to glycoside hydrolases and gives off information indicating just how active each of those enzymes is moment by moment.

Many of the measurements for the study, such as the measures of protein activity using mass spectrometry, were done at EMSL, the DOE’s Environmental Molecular Sciences Laboratory on the PNNL campus. Wright’s team included Lindsey Anderson, David Culley, Beth Hofstad, Lacie Chauvigné-Hines, Erika Zink, Samuel Purvine, Richard Smith, Stephen Callister, and Jon Magnuson, all of PNNL.

Resources

  • 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 (2013) “Activity-based protein profiling of secreted cellulolytic enzyme activity dynamics in Trichoderma reesei QM6a, NG14, and RUT-C30,” Molecular BioSystems doi: 10.1039/c3mb70333a

Comments

Gorr

,,make fuels from materials like switchgrass and corn stalks.,,

I want to buy these fuels but not from switchgrass or corn stalks but fron green algae harvest. There is not enouph quantities of switchgrass but green algae can be produced cheaper and in bigger quantities.

Insist that car manufacturers invest in green algae fuels and don't buy any new car from these manufacturers till you can buy cheaper biofuels at a pump near you. We strugglened enouph with high gas prices and poor mpg cars and suvs, it's now the time to begin a strike on new car sales.

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