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Expression of S-adenosylmethionine Hydrolase in Tissues Synthesizing Secondary Cell Walls Alters Specific Methylated Cell Wall Fractions and Improves Biomass Digestibility

机译:S-腺苷甲硫氨酸水解酶在组织中的表达合成次级细胞壁可改变特定的甲基化细胞壁部分并提高生物质消化率

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摘要

Plant biomass is a large source of fermentable sugars for the synthesis of bioproducts using engineered microbes. These sugars are stored as cell wall polymers, mainly cellulose and hemicellulose, and are embedded with lignin, which makes their enzymatic hydrolysis challenging. One of the strategies to reduce cell wall recalcitrance is the modification of lignin content and composition. Lignin is a phenolic polymer of methylated aromatic alcohols and its synthesis in tissues developing secondary cell walls is a significant sink for the consumption of the methyl donor S-adenosylmethionine (AdoMet). In this study, we demonstrate in Arabidopsis stems that targeted expression of AdoMet hydrolase (AdoMetase, E.C. 3.3.1.2) in secondary cell wall synthesizing tissues reduces the AdoMet pool and impacts lignin content and composition. In particular, both NMR analysis and pyrolysis gas chromatography mass spectrometry of lignin in engineered biomass showed relative enrichment of non-methylated p-hydroxycinnamyl (H) units and a reduction of dimethylated syringyl (S) units. This indicates a lower degree of methylation compared to that in wild-type lignin. Quantification of cell wall-bound hydroxycinnamates revealed a reduction of ferulate in AdoMetase transgenic lines. Biomass from transgenic lines, in contrast to that in control plants, exhibits an enrichment of glucose content and a reduction in the degree of hemicellulose glucuronoxylan methylation. We also show that these modifications resulted in a reduction of cell wall recalcitrance, because sugar yield generated by enzymatic biomass saccharification was greater than that of wild-type plants. Considering that transgenic plants show no important diminution of biomass yields, and that heterologous expression of AdoMetase protein can be spatiotemporally optimized, this novel approach provides a valuable option for the improvement of lignocellulosic biomass feedstock.
机译:植物生物量是使用工程微生物合成生物产品的可发酵糖的主要来源。这些糖以细胞壁聚合物(主要是纤维素和半纤维素)的形式存储,并被木质素包埋,这使其酶解具有挑战性。减少细胞壁顽固性的策略之一是木质素含量和组成的改变。木质素是甲基化芳族醇的酚类聚合物,其在形成次级细胞壁的组织中合成是甲基供体S-腺苷甲硫氨酸(AdoMet)消耗的重要吸收剂。在这项研究中,我们证明了拟南芥茎中在次级细胞壁合成组织中靶向表达AdoMet水解酶(AdoMetase,E.C. 3.3.1.2)会减少AdoMet库并影响木质素含量和组成。特别是,木质素在工程化生物质中的NMR分析和热解气相色谱质谱都显示非甲基化对羟基肉桂基(H)单元的相对富集和二甲基化丁香基(S)单元的减少。这表明与野生型木质素相比,甲基化程度较低。细胞壁结合的羟基肉桂酸酯的定量揭示了AdoMetase转基因系中阿魏酸盐的减少。与对照植物相比,来自转基因品系的生物质表现出丰富的葡萄糖含量和半纤维素葡糖醛酸木聚糖甲基化程度的降低。我们还表明,这些修饰导致细胞壁顽固性的降低,因为通过酶促生物质糖化作用产生的糖产量大于野生型植物的糖产量。考虑到转基因植物没有显示出生物量产量的重要减少,并且可以时空优化AdoMetase蛋白的异源表达,这种新颖的方法为改进木质纤维素生物量原料提供了有价值的选择。

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