Nanostructure-Initiator Mass Spectrometry (NIMS) for the Analysis of Enzyme Activities

Enzymes are essential to innumerable vital processes in all fields of life. Additionally, enzymatic activities are utilized in a multitude of biological, medical, and technical applications. Using cellulases and other glycoside hydrolases for the conversion of lignocellulosic biomass into biofuel has become an active area of research for applied enzymology. Development of high-throughput, high-specificity enzyme assays for glycoside hydrolases is critical to identify, characterize, and optimize such activities for efficient biomass degradation. Here, we present a detailed protocol describing our recently reported nanostructure-initiator mass spectrometry (NIMS)-based technique for the rapid analysis of glycoside hydrolase activities. NIMS enzyme activity (Nimzyme) assays can be used to characterize already known, or identify novel glycoside hydrolases. Importantly, several enzymatic activities can be tested in parallel in a multiplexed arrangement and under a broad range of assay conditions. Curr. Protoc. Chem. Biol. 4:123-142 © 2012 by John Wiley & Sons, Inc. Keywords: nanostructure-initiator mass spectrometry; NIMS; Nimzyme; enzyme assay; glycoside hydrolase; lignocellulosic biofuel