Enzymes play a central role in converting plant biomass into simple sugars and aromatic compounds that microbes can utilize to create sustainable fuels and bioproducts.
At the Joint BioEnergy Institute, we are helping expedite enzyme exploration. We are doing this using laser desorption based mass spectrometry (e.g. NIMS) for rapid detection of enzyme substrates and products including both activities of carbohydrate active enzymes and lignin modifying enzymes.
Recent highlights:
- Discovered highly specific ligninanse activities and products of lignin degradation
- Identified the substrate specificities and kinetics of novel multifunctional glycosyl hydrolases
This approach can allow for the high-throughput development of optimal biomass degrading cocktails. Our contributions together as part of the larger JBEI effort will help create cost-effective and sustainable bioenergy and bioproducts.
References:
Deng, K.; Zeng, J.; Cheng, G.; Gao, J.; Sale, K. L.; Simmons, B. A.; Singh, A. K.; Adams, P. D.; Northen, T. R. Rapid Characterization of the Activities of Lignin-Modifying Enzymes Based on Nanostructure-Initiator Mass Spectrometry (NIMS). Biotechnol. Biofuels 2018, 11 (1), 266. doi.org/10.1186/s13068-018-1261-2
Deng, K.; Guenther, J. M.; Gao, J.; Bowen, B. P.; Tran, H.; Reyes-Ortiz, V.; Cheng, X.; Sathitsuksanoh, N.; Heins, R.; Takasuka, T. E.; et al. Development of a High Throughput Platform for Screening Glycoside Hydrolases Based on Oxime-NIMS. Front. Bioeng. Biotechnol. 2015, 3. doi.org/10.3389/fbioe.2015.00153
Deng, K.; Takasuka, T. E.; Heins, R.; Cheng, X.; Bergeman, L. F.; Shi, J.; Aschenbrener, R.; Deutsch, S.; Singh, S.; Sale, K. L.; et al. Rapid Kinetic Characterization of Glycosyl Hydrolases Based on Oxime Derivatization and Nanostructure-Initiator Mass Spectrometry (NIMS). ACS Chem. Biol. 2014, 9 (7), 1470–1479. doi.org/10.1021/cb5000289
Northen, T. R.; Lee, J.-C.; Hoang, L.; Raymond, J.; Hwang, D.-R.; Yannone, S. M.; Wong, C.-H.; Siuzdak, G. A Nanostructure-Initiator Mass Spectrometry-Based Enzyme Activity Assay. Proc. Natl. Acad. Sci. 2008, 105 (10), 3678–3683. doi.org/10.1073/pnas.0712332105
Northen, T. R.; Yanes, O.; Northen, M. T.; Marrinucci, D.; Uritboonthai, W.; Apon, J.; Golledge, S. L.; Nordström, A.; Siuzdak, G. Clathrate Nanostructures for Mass Spectrometry. Nature 2007, 449 (7165), 1033–1036. doi.org/10.1038/nature06195