High throughput NIMS – Increasing the pace of functional genomics and synthetic biology
Technical advances in DNA sequencing and manipulation have led to an explosion in the sequenced universe and the ability to create libraries of millions or billions of bacterial variants. However, defining the functions of these remains slow and tedious. Our lab has a long-standing interest in nanostructure-initiator mass spectrometry (NIMS)1. We are using this in conjunction with nanoliter-scale manipulation of liquids for rapid mass spectrometry functional analysis2. We are particularly interested in the exoenzymes used by microorganisms to deconstruct plant and other biopolymers, including sugars, for subsequent uptake by cells. Through bioconjugate chemistry we modify these sugars enabling rapid detection and quantification using NIMS3,4. We are using this at the Joint BioEnergy Institute to help develop cellulosic biofuels both through screening metabolism and development of effective cocktails for deconstructing plant biomass.
1 Northen, T. R. et al. Clathrate nanostructures for mass spectrometry. Nature 449, 1033-1036, doi:10.1038/nature06195 (2007).
2 Greving, M. et al. Acoustic deposition with NIMS as a high-throughput enzyme activity assay. Analytical and bioanalytical chemistry 403, 707-711, doi:10.1007/s00216-012-5908-8 (2012).
3 Deng, K. et al. Encoding substrates with mass tags to resolve stereospecific reactions using Nimzyme. Rapid communications in mass spectrometry : RCM 26, 611-615, doi:10.1002/rcm.6134 (2012).
4 Northen, T. R. et al. A nanostructure-initiator mass spectrometry-based enzyme activity assay. Proc Natl Acad Sci U S A 105, 3678-3683, doi:10.1073/pnas.0712332105 (2008).