Characterization and Elimination of Undesirable Protein Residues in Plant Cell Wall Materials for Enhancing Lignin Analysis by Solution-State Nuclear Magnetic Resonance Spectroscopy,Biomacromolecules

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Characterization and Elimination of Undesirable Protein Residues in Plant Cell Wall Materials for Enhancing Lignin Analysis by Solution-State Nuclear Magnetic Resonance Spectroscopy
Biomacromolecules ( IF 5.5 ) Pub Date : 2017-11-08 00:00:00 , DOI: 10.1021/acs.biomac.7b01223
Hoon Kim _{1,

2} , Dharshana Padmakshan ₂ , Yanding Li _{2,

3} , Jorge Rencoret ₄ , Ronald D. Hatfield ₅ , John Ralph _{1,

2,

3}

Affiliation

Protein polymers exist in every plant cell wall preparation, and they interfere with lignin characterization and quantification. Here, we report the structural characterization of the residual protein peaks in 2D NMR spectra in corn cob and kenaf samples and note that aromatic amino acids are ubiquitous and evident in spectra from various other plants and tissues. The aromatic correlations from amino acid residues were identified and assigned as phenylalanine and tyrosine. Phenylalanine’s 3/5 correlation peak is superimposed on the peak from typical lignin p-hydroxyphenyl (H-unit) structures, causing an overestimation of the H units. Protein contamination also occurs when using cellulases to prepare enzyme lignins from virtually protein-free wood samples. We used a protease to remove the protein residues from the ball-milled cell walls, and we were able to reveal H-unit structures in lignins more clearly in the 2D NMR spectra, providing a better basis for their estimation.

中文翻译：

表征和消除植物细胞壁材料中不希望的蛋白质残基，以增强溶液状态核磁共振光谱法对木质素的分析

蛋白质聚合物存在于每种植物细胞壁制备物中，它们会干扰木质素的表征和定量。在这里，我们报告了玉米芯和洋麻样品的2D NMR光谱中残留蛋白质峰的结构特征，并注意到芳香族氨基酸无处不在，并且在来自其他植物和组织的光谱中也很明显。鉴定出来自氨基酸残基的芳族相关性，并将其分配为苯丙氨酸和酪氨酸。苯丙氨酸的3/5相关峰叠加在典型木质素p的峰上-羟苯基（H单元）结构，导致H单元的高估。当使用纤维素酶从几乎不含蛋白质的木材样品中制备酶木质素时，也会发生蛋白质污染。我们使用蛋白酶去除了球磨细胞壁上的蛋白质残基，并且我们能够在2D NMR光谱中更清楚地揭示木质素中的H单元结构，从而为它们的估计提供了更好的基础。

更新日期：2017-11-09

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