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首页> 外文期刊>Journal of Experimental Botany >Heterologous expression and characterization of an Arabidopsis beta-L-arabinopyranosidase and alpha-D-galactosidases acting on beta-L-arabinopyranosyl residues
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Heterologous expression and characterization of an Arabidopsis beta-L-arabinopyranosidase and alpha-D-galactosidases acting on beta-L-arabinopyranosyl residues

机译：拟南芥β-L-α-γ-阿拉伯吡喃糖苷酶和作用于β-1-阿拉伯吡喃糖基残基的α-D-半乳糖苷酶的异源表达及表征

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摘要

The major plant sugar l-arabinose (L-Ara) has two different ring forms, L-arabinofuranose (L-Araf) and l-arabinopyranose (L-Arap). Although L-Ara mainly appears in the form of alpha-L-Araf residues in cell wall components, such as pectic alpha-1,3:1,5-arabinan, arabinoxylan, and arabinogalactan-proteins (AGPs), lesser amounts of it can also be found as beta-L-Arap residues of AGPs. Even though AGPs are known to be rapidly metabolized, the enzymes acting on the beta-L-Arap residues remain to be identified. In the present study, four enzymes, which we call beta-L-ARAPASE (APSE) and alpha-GALACTOSIDASE 1 (AGAL1), AGAL2, and AGAL3, are identified as those enzymes that are likely to be responsible for the hydrolysis of the beta-L-Arap residues in Arabidopsis thaliana. An Arabidopsis apse-1 mutant showed significant reduction in beta-L-arabinopyranosidase activity, and an apse-1 agal3-1 double-mutant exhibited even less activity. The apse-1 and the double-mutants both had more beta-L-Arap residues in the cell walls than wild-type plants. Recombinant APSE expressed in the yeast Pichia pastoris specifically hydrolyzed beta-L-Arap residues and released L-Ara from gum arabic and larch arabinogalactan. The recombinant AGAL3 also showed weak beta-L-arabinopyranosidase activity beside its strong alpha-galactosidase activity. It appears that the beta-L-Arap residues of AGPs are hydrolysed mainly by APSE and partially by AGALs in Arabidopsis.
机译：主要植物糖L-阿拉伯糖（L-ARA）具有两种不同的环形形式，L-阿拉伯呋喃（L-ARAF）和L-阿拉伯吡喃糖（L- ARAP）。虽然L- ARA主要出现在细胞壁组分中α-L-ARAF残留物的形式，例如丙酸α-1,3：1,5-阿拉伯南，阿拉伯辛甲苯和阿拉伯半乳蛋白 - 蛋白（AGPS），其量较小也可以作为AGPS的β-L-ARAP残留物。即使已知迅速代谢AGPS，仍然仍然鉴定作用于β-L-ARAP残基的酶。在本研究中，我们称之为β-L- arapase（APSE）和α-半乳糖苷酶1（AgAl1），AgA12和AgAl3的四种酶被鉴定为可能负责β水解的酶-1-拟南芥的ARAP残留物。拟南芥APSE-1突变体表现出β-L-阿拉伯吡喃糖苷酶活性的显着降低，APSE-1 AgAl3-1双突变体表现出较少的活性。 APSE-1和双突变体均在细胞壁中具有比野生型植物更多的β-L-ARAP残基。在酵母Pichia Pastoris中表达的重组APSE特异性水解的β-L-ARAP残基，并释放来自阿拉伯语和甲羚羊醇的L- ara。重组AgAl3还显示出其强α-半乳糖苷酶活性旁边的β-L-阿拉伯吡喃糖苷酶活性。似乎AGPS的β-L-ARAP残基主要通过APSE水解，部分通过拟南芥中的AgAls水解。

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