Institute of Process Research and Development, School of Chemistry and School of Chemical and Process Engineering, University of Leeds, Woodhouse Lane, Leeds LS2 9JT, United Kingdom.
Institut Jean-Pierre Bourgin, INRAE, AgroParisTech, Université Paris-Saclay, Versailles 78000, France.
J Agric Food Chem. 2020 Dec 2;68(48):14297-14306. doi: 10.1021/acs.jafc.0c06308. Epub 2020 Nov 17.
A high-throughput agarose gel electrophoresis (AGE) analytical method has been developed to separate lignin fractions according to their molecular weight (), charge, and shape. Operating conditions to effect separation of species have been evaluated along with imaging parameters. Kraft, soda (Protobind), and Organosolv lignins showed distinct differences in migration. Bands were cut, extracted, and cross-analyzed by gel permeation chromatography (GPC), H NMR, and pyrolysis GC/MS to confirm their identity as lignin. The band intensity was correlated with lignin concentration by running serially diluted samples and imaging each lane to produce a precise calibration curve. The AGE technique was used to monitor and compare enzymatic, bacterial, chemical, and hydrothermal lignin digestions. Each method showed changes in lignin migration and band intensities over time. Low species were seen in samples collected from the anode buffer tank. Though requiring further development, the AGE method can provide structural information about the lignin and is accessible to biological and chemistry laboratories.
已开发出一种高通量琼脂糖凝胶电泳 (AGE) 分析方法，根据分子量 (), 电荷和形状分离木质素馏分。已评估了影响物种分离的操作条件和成像参数。 kraft、苏打 (Protobind) 和 Organosolv 木质素在迁移方面表现出明显差异。将条带切割、提取并通过凝胶渗透色谱 (GPC)、H NMR 和热解 GC/MS 进行交叉分析，以确认其为木质素。通过运行系列稀释样品并对每条泳道进行成像以产生精确的校准曲线，将条带强度与木质素浓度相关联。AGE 技术用于监测和比较酶促、细菌、化学和水热木质素消化。每种方法都显示出木质素迁移和条带强度随时间的变化。在从阳极缓冲罐收集的样品中观察到低 物种。尽管需要进一步开发，但 AGE 方法可以提供有关木质素的结构信息，并且易于生物学和化学实验室使用。
