Ben Elarbi Mosbah, Khemiri Halima, Jridi Taoufik, Ben Hamida Jeannette
Département de biologie, Tunis, Tunisia.
C R Biol. 2009 May;332(5):426-32. doi: 10.1016/j.crvi.2009.01.002. Epub 2009 Feb 27.
alpha-Amylase (alpha-1-4 D-glucan glucanohydrolase EC 3.2.1.1) crude extract was obtained from safflower (Carthamus tinctorius L.) cotyledons excised from 5-day-old dark grown seedlings. The enzyme was purified by precipitating the crude extract with ammonium sulphate at 20-60% saturation, and then by subjecting this fraction to affinity chromatography on a beta-cyclodextrin-Sepharose 6B column. The active fraction was dialysed and concentrated. An overall purification of about 131 folds with an activity yield of 81.25% was achieved. The molecular mass of purified enzyme determined by SDS-PAGE was 35 kD. When the purified alpha-amylase was subjected to gel electrophoresis followed by negative staining, only one band of active protein was detected. Its maximal activity was in the pH 6.0 and at a temperature of 55 degrees C. This enzyme was activated by Ca(2+) and inhibited by Fe(2+).
α-淀粉酶（α-1,4-D-葡聚糖葡聚糖水解酶，EC 3.2.1.1）粗提物取自5日龄黑暗培养幼苗的红花（Carthamus tinctorius L.）子叶。通过用20%-60%饱和度的硫酸铵沉淀粗提物，然后将该部分在β-环糊精-琼脂糖6B柱上进行亲和层析来纯化该酶。将活性部分透析并浓缩。实现了约131倍的总纯化，活性回收率为81.25%。通过SDS-PAGE测定的纯化酶的分子量为35 kD。当对纯化的α-淀粉酶进行凝胶电泳然后负染时，仅检测到一条活性蛋白带。其最大活性在pH 6.0和55℃温度下。该酶被Ca(2+)激活并被Fe(2+)抑制。
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