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Pectin Biosynthesis: GALS1 in Arabidopsis thaliana Is a β-14-Galactan β-14-Galactosyltransferase

机译:果胶生物合成:拟南芥中的GALS1是β-14-加拉坦β-14-半乳糖基转移酶

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摘要

β-1,4-Galactans are abundant polysaccharides in plant cell walls, which are generally found as side chains of rhamnogalacturonan I. Rhamnogalacturonan I is a major component of pectin with a backbone of alternating rhamnose and galacturonic acid residues and side chains that include α-1,5-arabinans, β-1,4-galactans, and arabinogalactans. Many enzymes are required to synthesize pectin, but few have been identified. Pectin is most abundant in primary walls of expanding cells, but β-1,4-galactan is relatively abundant in secondary walls, especially in tension wood that forms in response to mechanical stress. We investigated enzymes in glycosyltransferase family GT92, which has three members in Arabidopsis thaliana, which we designated GALACTAN SYNTHASE1, (GALS1), GALS2 and GALS3. Loss-of-function mutants in the corresponding genes had a decreased β-1,4-galactan content, and overexpression of GALS1 resulted in plants with 50% higher β-1,4-galactan content. The plants did not have an obvious growth phenotype. Heterologously expressed and affinity-purified GALS1 could transfer Gal residues from UDP-Gal onto β-1,4-galactopentaose. GALS1 specifically formed β-1,4-galactosyl linkages and could add successive β-1,4-galactosyl residues to the acceptor. These observations confirm the identity of the GT92 enzyme as β-1,4-galactan synthase. The identification of this enzyme could provide an important tool for engineering plants with improved bioenergy properties.
机译:β-1,4-半乳聚糖是植物细胞壁中丰富的多糖,通常被视为鼠李糖半乳糖醛酸聚糖I的侧链。鼠李糖半乳糖醛酸聚糖I是果胶的主要成分,具有交替的鼠李糖和半乳糖醛酸残基的骨架,且侧链包括α -1,5-阿拉伯聚糖,β-1,4-半乳聚糖和阿拉伯半乳聚糖。合成果胶需要许多酶,但很少有人发现。果胶在膨胀细胞的主壁中含量最高,但β-1,4-半乳聚糖在次壁中含量相对较高,特别是在响应机械应力而形成的张力木材中。我们调查了糖基转移酶家族GT92中的酶,该家族在拟南芥中有3个成员,我们将其命名为GALACTAN SYNTHASE1,(GALS1),GALS2和GALS3。相应基因中功能丧失的突变体的β-1,4-半乳聚糖含量降低,而GALS1的过表达导致植物的β-1,4-半乳聚糖含量增加50%。这些植物没有明显的生长表型。异源表达和亲和纯化的GALS1可以将Gal残基从UDP-Gal转移到β-1,4-半乳糖半乳糖上。 GALS1特异性形成β-1,4-半乳糖基键,并可以将连续的β-1,4-半乳糖基残基添加到受体上。这些观察结果证实了GT92酶作为β-1,4-半乳聚糖合酶的身份。这种酶的鉴定可以为工程植物的生物能源特性提供重要的工具。

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