Abstract

Lignocellulosic biomass is composed of three major biopolymers: cellulose, hemicellulose and lignin. Analytical tools capable of quickly detecting both glycan and lignin deconstruction are needed to support the development and characterization of efficient enzymes/enzyme cocktails. Previously we have described nanostructure-initiator mass spectrometry-based assays for the analysis of glycosyl hydrolase and most recently an assay for lignin modifying enzymes. Here we integrate these two assays into a single multiplexed assay against both classes of enzymes and use it to characterize crude commercial enzyme mixtures. Application of our multiplexed platform based on nanostructure-initiator mass spectrometry enabled us to characterize crude mixtures of laccase enzymes from fungi Agaricus bisporus (Ab) and Myceliopthora thermophila (Mt) revealing activity on both carbohydrate and aromatic substrates. Using time-series analysis we determined that crude laccase from Ab has the higher GH activity and that laccase from Mt has the higher activity against our lignin model compound. Inhibitor studies showed a significant reduction in Mt GH activity under low oxygen conditions and increased activities in the presence of vanillin (common GH inhibitor). Ultimately, this assay can help to discover mixtures of enzymes that could be incorporated into biomass pretreatments to deconstruct diverse components of lignocellulosic biomass.

Details

Title
A multiplexed nanostructure-initiator mass spectrometry (NIMS) assay for simultaneously detecting glycosyl hydrolase and lignin modifying enzyme activities
Author
Ing, Nicole 1 ; Deng Kai 1 ; Chen, Yan 2 ; Aulitto Martina 2 ; Gin, Jennifer W 2 ; Pham Thanh Le Mai 1 ; Petzold, Christopher J 2 ; Singer, Steve W 2 ; Bowen, Benjamin 3 ; Sale, Kenneth L 1 ; Simmons, Blake A 2 ; Singh, Anup K 1 ; Adams, Paul D 4 ; Northen, Trent R 2 

 Joint BioEnergy Institute, Emeryville, USA (GRID:grid.451372.6) (ISNI:0000 0004 0407 8980); Sandia National Laboratories, Livermore, USA (GRID:grid.474523.3) (ISNI:0000000403888279) 
 Joint BioEnergy Institute, Emeryville, USA (GRID:grid.451372.6) (ISNI:0000 0004 0407 8980); Lawrence Berkeley National Laboratory, Berkeley, USA (GRID:grid.184769.5) (ISNI:0000 0001 2231 4551) 
 Lawrence Berkeley National Laboratory, Berkeley, USA (GRID:grid.184769.5) (ISNI:0000 0001 2231 4551) 
 Joint BioEnergy Institute, Emeryville, USA (GRID:grid.451372.6) (ISNI:0000 0004 0407 8980); Lawrence Berkeley National Laboratory, Berkeley, USA (GRID:grid.184769.5) (ISNI:0000 0001 2231 4551); University of California, Berkeley, USA (GRID:grid.47840.3f) (ISNI:0000 0001 2181 7878) 
Publication year
2021
Publication date
2021
Publisher
Nature Publishing Group
e-ISSN
20452322
Source type
Scholarly Journal
Language of publication
English
ProQuest document ID
2536653326
Copyright
© The Author(s) 2021. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.