绵农小麦种子巯基蛋白酶抑制剂的纯化及性质研究
, PP. 140-147
Keywords: 巯基蛋白酶抑制剂,小麦巯基蛋白酶抑制剂,小麦巯基蛋白酶抑制剂的性质
Abstract:
绵农11号小麦种子磨粉后的胚和麸皮,经溶液浸取和硫酸铵分级沉淀获得巯基蛋白酶抑制剂(cpi)粗品,再经木瓜蛋白酶-sepharose4b亲和层析,deae-sepharose离子交换层析和sephadexg-100分子筛层析,可获得两种cpi.通过sds-page或凝胶过滤法测得其mr分别为18800[简称cpi(h)]和10500[简称cpi(l)].它们在page,sds-page和hplc上均为单一蛋白带。cpi(l)和cpi(h)对木瓜白酶的抑制活性(id50)分别为7.8×10-6mol/l和3.40×10-6mol/l.对无花果蛋白酶的抑制活性分别为9.5×10-5mol/l和5.8×10-6mol/l;cpi(h)对菠萝蛋白酶有弱抑制作用,但cpi(l)则无抑制作用;无论是cpi(l)或cpi(h)对胰蛋白酶、胰凝乳蛋白酶和胃蛋白酶均无抑制作用。cpi(h)的n-末端为ile.cpi(h)和cpi(l)在ph2.0～11.0范围内和90℃处理5min,两种cpi的抑制活性均无变化;cpi(h)和cpi(l)对木爪蛋白酶的抑制曲线经dixon作图法,得出cpi(h)为反竞争性抑制类型,而cpi(l)为竞争性抑制类型,其ki值为8.32×10-6mol/l.
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