水稻RuBP羧化酶的提纯及电泳性质分析

RuBP(1、5—二磷酸核酮础)羧化酶,在绿色植物的光合过程中起着重要作用。我们采用Sephadex 柱层析法从水稻中分离和纯化了该酶,并获得了结晶。经电泳鉴定为一条酶带。该酶结品又经解聚,S—羧甲基化和聚丙烯酰胺凝胶等电聚焦电泳分析呈5条带,3条为大亚基肽链,2条为小亚基肽链。     

等电聚焦强度对木薯叶片蛋白质双向电泳分离结果的影响

通过优化等电聚焦强度改良木薯叶片蛋白质双向电泳的分离效果。 结果表明,13 万 Vhs 等电聚焦强度能很好地分离木薯叶片蛋白质, 且分离的蛋白质具有良好的质谱兼容性。质谱鉴定 7 个蛋白点, 结果显示这些蛋白主要参与能量代谢,其中有些蛋白点是在 13 万 Vhs 聚焦强度下特异出现的     

Laccase and peroxidase isoenzymes during leaf litter decomposition of Quercus ilex in a Mediterranean ecosystem

The dynamics of leaf litter decomposition of Quercus ilex (L.) were investigated over a 2 year period by determining the activities and isoenzyme distribution of laccases and peroxidases. The analysis of isoenzymes was performed by isoelectric focusing on high stability pH gradients with high resolving power. The preparation of zymograms was carried out using the leaf litter extract without previous concentration. During litter decomposition, laccase and peroxidase activities changed as well as the type and number of enzyme isoforms. The activities of both enzymes were low (≤0.017 and ≤0.031 mmol o-tolidine oxidized h⁻¹ g⁻¹ d.w. for laccase and peroxidase, respectively) in first year and increased in October-January of the second year of litter decay. The highest activities measured after 15-18 months of litter exposure (0.37±0.03 and 0.19±0.02 mmol o-tolidine oxidized h⁻¹ g⁻¹ d.w. for laccase and peroxidase, respectively), showed that litter chemical composition affected the growth of ligninolytic microbial community. The activation energy for laccase and peroxidase reactions also changed during decomposition: the highest values (55±6 kJ mol⁻¹ for laccase and 60±6 kJ mol⁻¹ for peroxidase) occurred in autumn-winter, even if spatial changes were evidenced. Some enzyme isoforms (pI=5.3 and 5.5 for laccase and pI=5.0 and 5.1 for peroxidase, respectively), contributed more than others to the overall laccase and peroxidase activity, suggesting that some ligninolytic species bloomed in particular seasons of the year, even if other species with similar functional activities colonized the litter.