- Biofuel production
- Drug discovery
- Enzyme bioengineering
- Allows native substrates to screen for specific enzymatic activities
- Small sample volume, high throughput
- Potential to aid explanation of enzyme catalytic mechanism
Creating optimal enzymes to improve the efficiency of converting biomass into sugars for biofuel production requires efficient screens to provide information on enzyme specificity and activity. Researchers at the Joint BioEnergy Institute (JBEI) have developed a technology to create a more efficient workflow for hydrolytic enzyme discovery and enzyme cocktail optimization by providing fast, efficient analysis of native glycans using high specificity mass spectrometry-based enzyme assays.
In the JBEI technology, native substrates are used for enzyme activity screening and then tagged for efficient mass spectrometry analysis. The tagged mixture is assayed using mass spectrometry-based arrays that enable high throughput screening from microwell plates. Integrating this technology with acoustic printing has yielded extremely high throughput (three minute / 384 well plate) mass spectrometry arrays.
In the case of hydrolytic enzyme library screening, if incomplete hydrolysis is observed, the mixture can be screened for additional enzymes that would complete the hydrolysis. As enzymes are added, the mixture can be used to screen for other enzymes to add to the cocktail until the desired conversion of biomass is achieved.
DEVELOPMENT STAGE: Proven principle. Researchers have tested and characterized 1) glycohydrolase activities in ionic liquid pretreated switchgrass and AFEX pretreated switchgrass, and 2) glycotransferase activities.
STATUS: Patent pending. Available for licensing or collaborative research.
REFERENCE NUMBER: EJIB-3262
The Joint BioEnergy Institute (JBEI, www.jbei.org) is a scientific partnership led by the Lawrence Berkeley National Laboratory and including the Sandia National Laboratories, the University of California campuses of Berkeley and Davis, the Carnegie Institution for Science and the Lawrence Livermore National Laboratory. JBEI’s primary scientific mission is to advance the development of the next generation of biofuels.