Hernández-Saavedra N Y, Ochoa J L
Center for Biological Research, Northwest, Laboratory of Marine Yeast, Baja California Sur, México.
Yeast. 1999 Jun 15;15(8):657-68. doi: 10.1002/(SICI)1097-0061(19990615)15:8<657::AID-YEA410>3.0.CO;2-U.
We have isolated the cytosolic form of Cu-Zn superoxide dismutase (SOD) from the marine yeast Debaryomyces hansenii. This enzyme has a subunit mass of 18 kDa. The preparation was found to be heterogeneous by IF electrophoresis with two pI ranges: 5.14-4.0 and 1.6-1.8. The enzyme preparation had a remarkably strong stability at pH 6.0-7.0, surviving boiling for 10 min without losing more than 60% of activity. On Western blots, this enzyme was recognized by antibodies raised in rabbits against D. hansenii extracts, while only a weak cross-reaction could be detected using antibodies generated against either Saccharomyces cerevisiae or bovine erythrocyte Cu-Zn SODs. In sequencing analysis, a peptide obtained by trypsin digestion was found to have 85% identity to the S. cerevisiae Cu-Zn SOD.
我们从海洋酵母汉逊德巴利酵母中分离出了胞质形式的铜锌超氧化物歧化酶(SOD)。这种酶的亚基质量为18 kDa。通过等聚焦电泳发现该制剂具有异质性,有两个pI范围:5.14 - 4.0和1.6 - 1.8。该酶制剂在pH 6.0 - 7.0时具有非常强的稳定性,煮沸10分钟后活性损失不超过60%。在蛋白质免疫印迹分析中,这种酶能被用兔抗汉逊德巴利酵母提取物产生的抗体识别,而使用抗酿酒酵母或牛红细胞铜锌SOD产生的抗体时只能检测到微弱的交叉反应。在测序分析中,发现通过胰蛋白酶消化获得的一个肽段与酿酒酵母铜锌SOD有85%的同源性。