Froman D P
Department of Poultry Science, Oregon State University, Corvallis 97331.
Poult Sci. 1990 May;69(5):812-7. doi: 10.3382/ps.0690812.
The objective of the present research was to identify a procedure whereby chicken acrosin could be purified. Acrosin, as evidenced by amidase activity, was extracted with urea most efficiently at a concentration of 6 M. Extraction efficiency was enhanced by spermatozoal lysis prior to admixture with 6 M urea. Lysis was induced by passage of spermatozoal suspensions through a French pressure cell. Acrosin was purified by using gel filtration, chromatofocusing, and affinity chromatography. Based on amidase activity, a 19-fold purification was obtained with a 28% recovery. Native electrophoresis resolved two major protein bands with proteolytic activity. The methods described afford the procurement of milligram amounts of chicken acrosin.
本研究的目的是确定一种纯化鸡顶体蛋白酶的方法。通过酰胺酶活性证明,在6M浓度的尿素中提取顶体蛋白酶效率最高。在与6M尿素混合之前,精子溶解可提高提取效率。精子悬浮液通过法国压力细胞诱导溶解。通过凝胶过滤、层析聚焦和亲和层析纯化顶体蛋白酶。基于酰胺酶活性,获得了19倍的纯化,回收率为28%。天然电泳解析出两条具有蛋白水解活性的主要蛋白带。所述方法可获取毫克量的鸡顶体蛋白酶。