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首页> 外文期刊>Biochimica et biophysica acta: international journal of biochemistry and biophysics >Enzymatic properties of endo-beta-N-acetylglucosaminidases from developing tomato fruits and soybean seeds: substrate specificity of plant origin endoglycosidase.
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Enzymatic properties of endo-beta-N-acetylglucosaminidases from developing tomato fruits and soybean seeds: substrate specificity of plant origin endoglycosidase.

机译:来自发育中的番茄果实和大豆种子的内切β-N-乙酰氨基葡萄糖苷酶的酶学性质:植物来源的内切糖苷酶的底物特异性。

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摘要

Substrate specificity and some other enzymatic properties of partial purified endo-beta-N-acetylglucosaminidases (endo-beta-GlcNAc-ase) from developing soybean seeds (Glycine max, Endo-GM) and developing tomato fruits (Lycopersicum esculentum, Endo-LE) were studied. The substrate specificity of these two endoglycosidases was explored and compared with regard to various pyridylaminated N-glycans derived from some naturally occurring glycoproteins. For Endo-GM and Endo-LE, several high mannose-type sugar chains bearing alpha 1-2 mannosyl residue(s), Man9-6GlcNAc2-PA (PA is pyridylamino) (80-100% relative hydrolysis), were most favored substrates followed by Man5GlcNAc2-PA (32% for Endo-LE, 43% for Endo-GM), a typical hybrid-type structure (GlcNAc1Man5GlcNAc2-PA; 34% for Endo-LE, 37% for Endo-GM), and then the common core pentasaccharide of N-glycan (Man3GlcNAc2-PA; 9% for Endo-GM and 16% for Endo-LE). On the contrary, both Endo-GM and Endo-LE could barely hydrolyze the xylose-containing N-glycans (Man3Xyl1GlcNAc2-PA, Man3Fuc1Xyl1GlcNAc2-PA) found ubiquitously in plant cells. The molecular mass of these two endoglycosidases was approximately 62 kDa by gel filtration and both Endo-GM and Endo-LE showed maximal activities for Man6GlcNAc2-PA in a weak acidic region (pH 6.0-6.5).
机译:发育中的大豆种子(Glycine max,Endo-GM)和发育中的番茄果实(Lycopersicum esculentum,Endo-LE)的部分纯化的内源β-N-乙酰氨基葡萄糖苷酶(内源-β-GlcNAc-酶)的底物特异性和其他一些酶学特性被研究了。探索了这两种内切糖苷酶的底物特异性,并与衍生自某些天然存在的糖蛋白的各种吡啶化的N-聚糖进行了比较。对于Endo-GM和Endo-LE,几条带有α1-2甘露糖基残基的高甘露糖型糖链,Man9-6GlcNAc2-PA(PA为吡啶基氨基)(相对水解80-100%)是最优选的底物其次是Man5GlcNAc2-PA(Endo-LE为32%,Endo-GM为43%),典型的混合型结构(GlcNAc1Man5GlcNAc2-PA; Endo-LE为34%,Endo-GM为37%),然后是N-聚糖的常见核心五糖(Man3GlcNAc2-PA; Endo-GM为9%,Endo-LE为16%)。相反,Endo-GM和Endo-LE都几乎不能水解植物细胞中普遍存在的含木糖的N-聚糖(Man3Xyl1GlcNAc2-PA,Man3Fuc1Xyl1GlcNAc2-PA)。通过凝胶过滤,这两种糖苷内切酶的分子量约为62 kDa,Endo-GM和Endo-LE在弱酸性区域(pH 6.0-6.5)中均显示出Man6GlcNAc2-PA的最大活性。

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