摘要
用小玻璃珠破碎处于对数生长中期的粟酒裂殖酵母（Ｓｃｈｉｚｏｓａｃｃｈａｒｏｍｙｃｅｓｐｏｍｂｅ）细胞，将所得到的匀浆液经差速离心，酸处理，不连续蔗糖密度梯度离心，得到质膜ＡＴＰ酶，并对其进行鉴定及活性测定．发现所制备的质膜ＡＴＰ酶对ＮａＮ３不敏感，而对Ｎａ３ＶＯ４具有高度的敏感性，半抑制浓度为１２μｍｏｌ／Ｌ，表明所制备的质膜ＡＴＰ酶较纯，不混杂线粒体ＡＴＰ酶．该酶的最适ｐＨ为６．３，对Ｍｇ２＋有很强的依赖性，对ＡＴＰ有较强的亲和性，Ｋｍ值为１．３ｍｍｏｌ／Ｌ．
The cells of
Schizosaccharomyces pombe which were in mid-exponestial growth phase were homogenized
with glass beads. The plasma membrane ATPases were obtained by differential centrifugation,
followed by acid treatment and isopycnic centrifugation in a discontinuous sucrose gradient,
then their activities were assayed. It was defected that the preparation of plasma membrane
ATPases was insentive to NaN 3,but highly sensitive to Na 3VO 4 with an IC 50 ≈12μ
mol/L. It wasverified that the preparation of plasma membrane ATPases was purified and didn't
contain mitochondrial ATPases.The pH optimum of the plasma membrane ATPases was 6.3.
The ATPases highly depended on Mg 2+ and showed a high affinity for ATP, Km=1.3mmol/L.
出处
《南京师大学报（自然科学版）》
CAS
CSCD
1999年第2期83-87,共5页
Journal of Nanjing Normal University(Natural Science Edition)
基金
江苏省自然科学基金
关键词
粟酒裂殖酵母
质膜
ATP酶
活性
测定
酿酒酵母
Schizosaccharomyces pombe ,plasma membrane ATPase, preparation , assay
activity
