A multiplexed nanostructure-initiator mass spectrometry (NIMS) assay for simultaneously detecting glycosyl hydrolase and lignin modifying enzyme activities

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. Previous...

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Autores principales:	Nicole Ing, Kai Deng, Yan Chen, Martina Aulitto, Jennifer W. Gin, Thanh Le Mai Pham, Christopher J. Petzold, Steve W. Singer, Benjamin Bowen, Kenneth L. Sale, Blake A. Simmons, Anup K. Singh, Paul D. Adams, Trent R. Northen
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/80b91501f88e4390b6e8f4cc7fce402b
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spelling	oai:doaj.org-article:80b91501f88e4390b6e8f4cc7fce402b2021-12-02T15:02:31ZA multiplexed nanostructure-initiator mass spectrometry (NIMS) assay for simultaneously detecting glycosyl hydrolase and lignin modifying enzyme activities10.1038/s41598-021-91181-82045-2322https://doaj.org/article/80b91501f88e4390b6e8f4cc7fce402b2021-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-91181-8https://doaj.org/toc/2045-2322Abstract 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.Nicole IngKai DengYan ChenMartina AulittoJennifer W. GinThanh Le Mai PhamChristopher J. PetzoldSteve W. SingerBenjamin BowenKenneth L. SaleBlake A. SimmonsAnup K. SinghPaul D. AdamsTrent R. NorthenNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Nicole Ing Kai Deng Yan Chen Martina Aulitto Jennifer W. Gin Thanh Le Mai Pham Christopher J. Petzold Steve W. Singer Benjamin Bowen Kenneth L. Sale Blake A. Simmons Anup K. Singh Paul D. Adams Trent R. Northen A multiplexed nanostructure-initiator mass spectrometry (NIMS) assay for simultaneously detecting glycosyl hydrolase and lignin modifying enzyme activities
description	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.
format	article
author	Nicole Ing Kai Deng Yan Chen Martina Aulitto Jennifer W. Gin Thanh Le Mai Pham Christopher J. Petzold Steve W. Singer Benjamin Bowen Kenneth L. Sale Blake A. Simmons Anup K. Singh Paul D. Adams Trent R. Northen
author_facet	Nicole Ing Kai Deng Yan Chen Martina Aulitto Jennifer W. Gin Thanh Le Mai Pham Christopher J. Petzold Steve W. Singer Benjamin Bowen Kenneth L. Sale Blake A. Simmons Anup K. Singh Paul D. Adams Trent R. Northen
author_sort	Nicole Ing
title	A multiplexed nanostructure-initiator mass spectrometry (NIMS) assay for simultaneously detecting glycosyl hydrolase and lignin modifying enzyme activities
title_short	A multiplexed nanostructure-initiator mass spectrometry (NIMS) assay for simultaneously detecting glycosyl hydrolase and lignin modifying enzyme activities
title_full	A multiplexed nanostructure-initiator mass spectrometry (NIMS) assay for simultaneously detecting glycosyl hydrolase and lignin modifying enzyme activities
title_fullStr	A multiplexed nanostructure-initiator mass spectrometry (NIMS) assay for simultaneously detecting glycosyl hydrolase and lignin modifying enzyme activities
title_full_unstemmed	A multiplexed nanostructure-initiator mass spectrometry (NIMS) assay for simultaneously detecting glycosyl hydrolase and lignin modifying enzyme activities
title_sort	multiplexed nanostructure-initiator mass spectrometry (nims) assay for simultaneously detecting glycosyl hydrolase and lignin modifying enzyme activities
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/80b91501f88e4390b6e8f4cc7fce402b
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A multiplexed nanostructure-initiator mass spectrometry (NIMS) assay for simultaneously detecting glycosyl hydrolase and lignin modifying enzyme activities

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