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测得果菠萝蛋白酶是由单亚基组成,分子量约为30k,pl为4.6,每个酶分子含一个游离疏基和二个硫硫键,甲醇、乙醇、乙二醇对酶活力有明显的抑制作用,甲醇、乙醇表现为非竞争性,乙二醇为竞争性,其抑制常数分别为:20%、16.5%和11%。在有机溶剂作用下,监测荧光(332nm)强度随时间的变化过程,作为变性的指标,测定酶在不同浓度有机溶剂中的变性动力学,并按邹氏法测定酶的失活动力学,对酶构象及催化活力关系进行比较,酶的失活速度大于变性速度,说明酶活性中心处于对有机溶剂较敏感的区域,酶的变性与失活过程均可分为快、慢两相的一级反应,这意味着可能存在着部分活力的中间态。
The measured results bromelain is composed of a single subunit, molecular weight of about 30k, pl 4.6, each enzyme molecule contains a free sparse and two sulfur-sulfur bond, methanol, ethanol, ethylene glycol on the enzyme activity was significant Inhibition, methanol, ethanol showed noncompetitive, ethylene glycol is competitive, the inhibition constants were: 20%, 16.5% and 11%. Under the action of organic solvents, the intensity of fluorescence (332nm) was monitored over time. The denaturation kinetics of the enzyme in different concentrations of organic solvents was determined as an indicator of denaturation. The kinetics of enzyme inactivation was determined by Zou’s method. Conformational and catalytic vitality, the rate of enzyme inactivation is greater than the denaturation rate, indicating that the enzyme activity center is sensitive to organic solvents in the region, the enzyme denaturation and inactivation process can be divided into fast and slow two-phase reaction , Which means that there may be some vitality of the intermediate state.