Reconstructing the chemical and structural characteristics of the plant cell wall represents a promising solution to overcoming lignocellulosic biomass recalcitrance to biochemical deconstruction. This study aims to leverage hydroxyproline (Hyp)-O-glycosylation, a process unique to plant cell wall glycoproteins, as an innovative technology for de novo design and engineering in planta of Hyp-O-glycosylated biopolymers (HypGP) that facilitate plant cell wall reconstruction. HypGP consisting of 18 tandem repeats of "Ser-Hyp-Hyp-Hyp-Hyp" motif or (SP4)18 was designed and engineered into tobacco plants as a fusion peptide with either a reporter protein enhanced green fluorescence protein or the catalytic domain of a thermophilic E1 endoglucanase (E1cd) from Acidothermus cellulolyticus. The engineered (SP4)18 module was extensively Hyp-O-glycosylated with arabino-oligosaccharides, which facilitated the deposition of the fused protein/enzyme in the cell wall matrix and improved the accumulation of the protein/enzyme in planta by 1.5-11-fold. The enzyme activity of the recombinant E1cd was not affected by the fused (SP4)18 module, showing an optimal temperature of 80°C and optimal pH between 5 and 8. The plant biomass engineered with the (SP4)18 -tagged protein/enzyme increased the biomass saccharification efficiency by up to 3.5-fold without having adverse impact on the plant growth.

中文翻译：

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工程化羟基脯氨酸-O-糖基化生物聚合物，以重建植物细胞壁，从而提高生物质的可加工性。

重建植物细胞壁的化学和结构特征是克服木质纤维素生物质对生化解构的顽强抵抗的有前途的解决方案。这项研究旨在利用羟脯氨酸（Hyp）-O-糖基化这一植物细胞壁糖蛋白特有的过程，作为一种创新的技术，从头设计和工程化植物中的Hyp-O-糖基化生物聚合物（HypGP），从而促进植物细胞壁。重建。由“ Ser-Hyp-Hyp-Hyp-Hyp”基序或（SP4）18的18个串联重复序列组成的HypGP被设计和改造成烟草植物，是一种融合肽，与报告蛋白增强的绿色荧光蛋白或催化结构域相关。嗜酸嗜热纤维菌的嗜热E1内切葡聚糖酶（E1cd）。经过工程改造的（SP4）18模块被阿拉伯寡糖广泛地Hyp-O-糖基化，这有助于融合蛋白/酶在细胞壁基质中的沉积，并使蛋白/酶在植物体内的积累提高了1.5-11-折。重组E1cd的酶活性不受融合的（SP4）18模块的影响，显示的最佳温度为80°C，最佳pH在5至8之间。用（SP4）18标签的蛋白/酶工程化的植物生物量。将生物质的糖化效率提高了3.5倍，而对植物的生长没有不利影响。