{"title":"NMR spectroscopy: An invaluable tool to identify lignin structural modifications induced by oxidative enzymes.","authors":"Nicolò Pajer, Umberto Danelon, Claudia Crestini","doi":"10.1016/bs.mie.2025.02.003","DOIUrl":null,"url":null,"abstract":"<p><p>This chapter provides a comprehensive guide to the acquisition, processing, and analysis of NMR data for investigating the behaviour of lignins in enzymatic treatment. The necessary pre-treatments to obtain purified lignin samples, free from enzymes and contaminants, are described prior to the spectroscopic analyses. Detailed instructions for using quantitative <sup>13</sup>C NMR with an internal standard are then given to determine the abundance of lignin bonding patterns, expressed both as mmol/g of lignin and normalised on the basis of an aromatic unit. Quantitative <sup>31</sup>P NMR is the favoured analytical tool used to distinguish and quantify hydroxyl groups, which are key targets for oxidases. Finally, both qualitative and quantitative HSQC analyses (QQ-HSQC and HSQC<sub>0</sub>) are described to monitor overall changes in lignin bonding patterns.</p>","PeriodicalId":18662,"journal":{"name":"Methods in enzymology","volume":"716 ","pages":"33-104"},"PeriodicalIF":0.0000,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Methods in enzymology","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1016/bs.mie.2025.02.003","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2025/2/26 0:00:00","PubModel":"Epub","JCR":"Q3","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
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Abstract
This chapter provides a comprehensive guide to the acquisition, processing, and analysis of NMR data for investigating the behaviour of lignins in enzymatic treatment. The necessary pre-treatments to obtain purified lignin samples, free from enzymes and contaminants, are described prior to the spectroscopic analyses. Detailed instructions for using quantitative 13C NMR with an internal standard are then given to determine the abundance of lignin bonding patterns, expressed both as mmol/g of lignin and normalised on the basis of an aromatic unit. Quantitative 31P NMR is the favoured analytical tool used to distinguish and quantify hydroxyl groups, which are key targets for oxidases. Finally, both qualitative and quantitative HSQC analyses (QQ-HSQC and HSQC0) are described to monitor overall changes in lignin bonding patterns.
期刊介绍：
The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.
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