摘要
甲基营养菌WB 1菌株在以甲胺磷作碳氮源的无机盐培养基上生长时 ,产生甲胺磷降解酶情况较好 ,培养 2 0h为细胞收获的适宜时期。所得细胞经过超声波破碎、吐温 2 0抽提、热处理 ( 60℃ ,9min)、DEAE 纤维素 32和CM 纤维素 32柱层析等步骤 ,得到的酶制品比活力为 1 8 0U/mg ,收率 78 8% ,纯化倍数 2 2 8。纯化的酶制品经连续聚丙烯酰胺凝胶电泳 ,呈现一条主带 ,酶活力染色则呈现一条与之对应的活性谱带 ;该酶催化反应的适宜 pH为9 0 ,底物专一性不强 ,Hg2 +、Mn2 +、Cu2 +对酶有强烈的抑制作用。部分纯化酶制品在 - 2 0℃( 0 .1mol/L的磷酸盐溶液中 )存放 5d ,酶活力丧失约 60 %。
After cultivation for 20h in the inorganic salt medium with methamidophos as sole carbon and nitrogen source, Methylotroph WB 1 could produce methamidophos\|degrading enzyme in larger amounts. The enzyme was partially purified by sonication disruption, Tween\|20 extraction, heat treatment(9?min at 60℃), DEAE\|cellulose 32 and CM\|cellulose32 chromatography with 22.8 times purification and 78.8% recovery. Activity staining showed single violate band corresponding to that of protein staining. Optimum pH of the enzyme was 9.0; poor substrate specificity was showed. It was strongly inhibited by Hg 2+ \,Mn 2+ and Cu 2+ , but not by Co 2+ and Zn 2+ ; it also exhibited poor stability.
出处
《微生物学报》
CAS
CSCD
北大核心
2000年第5期523-527,共5页
Acta Microbiologica Sinica
基金
国家自然科学基金资助项目!( 3 9670 0 1 0 )
"863"计划资助项目!(SZ -0 3 -0 1 -0 3 )&&
关键词
甲基营养酶
甲胺磷降解酶
纯化
Methylotroph bacterium, Methamidophos\|degrading enzyme, Purification
