Study of the mode of action of endopolygalacturonase from Fusarium moniliforme.
Study of the mode of action of endopolygalacturonase from Fusarium moniliforme.
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串珠镰刀菌内聚半乳糖醛酸酶作用方式的研究。
DOI：
10.1016/s0304-4165(01)00141-6
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发表时间：
2001
期刊：
影响因子：
--
通讯作者：
J. Thibault
中科院分区：
文献类型：
--
作者：
E. Bonnin;A. L. Le Goff;R. Körner;G. W. Van Alebeek;T. Christensen;A. Voragen;P. Roepstorff;C. Caprari;J. Thibault
从酿酒酵母转化菌株的培养液中纯化出一种来自串珠镰刀菌的内聚半乳糖醛酸酶。在半乳糖醛酸低聚物和高聚半乳糖醛酸上研究了其动力学参数和作用方式。二聚体不是酶的底物。该酶对其他测试底物表现出米氏行为。亲和力和最大水解速率随着链长度的增加而增加，直至六聚体或七聚体，其Vmax与同型半乳糖醛酸处于相同范围内。该酶被证明具有多链攻击作用模式，其活性位点包括从-3到+2的五个子位点。高半乳糖醛酸水解的最终产物是半乳糖醛酸单体和二聚体。
One endopolygalacturonase from Fusarium moniliforme was purified from the culture broth of a transformed strain of Saccharomyces cerevisiae. Its kinetic parameters and mode of action were studied on galacturonic acid oligomers and homogalacturonan. The dimer was not a substrate for the enzyme. The enzyme was shown to follow Michaelis–Menten behaviour towards the other substrates tested. Affinity and maximum rate of hydrolysis increased with increasing chain length, up to the hexamer or heptamer, for which Vmaxwas in the same range as with homogalacturonan. The enzyme was demonstrated to have a multi-chain attack mode of action and its active site included five subsites ranging from −3 to +2. The final products of hydrolysis of homogalacturonan were the monomer and the dimer of galacturonic acid.