甲氰菊酯降解酶的部分纯化及其性质研究

Studies on Partial Purification of Fenpropathrin Degrading Enzyme and Its Characterization

恶臭假单胞菌CP-1可以在甲氰菊酯作为碳、氮源的无机盐培养基上生长,培养60 h时,可以产生最大量的甲氰菊酯降解酶,降解酶活力达到约7.5 U/ml。该酶通过离子交换柱层析,纯化了5.67倍,比活力达到85.0U/mg,回收率为90.6%。经SDS-PAGE凝胶电泳,酶蛋白染色呈现一条主带,该酶的分子量约为78kD。等电聚焦测得酶的等电点约为6.5。冻干后的酶保存于4℃冰箱1个月,对甲氰菊酯的降解活力为原有活力的63%。

After cultivation for 60h in the inorganic salt medium with fenpropathrin as sole carbon and nitrogen source, Pseudomonas putida CP-1 could produce fenpropathrin-degrading enzyme in largest amounts, The vigor of degrading enzyme attained 7,5 U/ml. The enzyme was partially purified by the ion exchange, DEAE-cellulose 52 and CM- cellulose 52 chromatography, with 5.67 times purification and 90.6 % recovery. The enzyme attained the specific activity of 85.0U/mg. With SDS-PAGE electrophoresis, the protein presented as a dark band. Compared with the protein marker, the molecular weight of the enzyme Was roughly 78kD. The IEF-PAGE measurement showed pI of the enzyme was 6, 5. The enzyme had 63 % of the total degradation activity after one month restoration at 4℃.