嗜水气单胞菌胞外蛋白酶ECPase54的纯化及特性分析.pdfVIP

199 6 , 19 ( 3) : 8 8 9 4 J ournal of N anj ing A g ric ult ur al Un iv er sity ECPase 54 李焕荣　陈怀青　陆承平* * ( , 210095) ( ) -1 56 ( ) Ah J EDT A ( PMSF) , (ECPase) , Ah J-1 SDS, 37 , , 5 -1 - -200 , Ah J DEAE Se hadex G 2-, 54 000, 2-, 35 000, , EDTA , PM SF ( TSMP) , ECPase 54 H 7. 5, ( Km) 1.77 m g/ ml, Vero , ; ; ; S852. 61 PURIFICATION AND CHARACTERIZATION OF EXTRACELLULAR PROTEASE ECPASE 54 FROM AEROMONAS HYDROPHILA Li Huanrong, Chen Huaiqing and Lu Cheng ing ( , , 210095) College of Veterinary M edicine Nanjing Agric Univ Nanjing ( ) -1 ABSTRACT At least five extracellular roteases ECPase in A eromonas hy d rop h ila strain J culture su- - 0. 1% . ernatant w ere assayed by in situ roteolytic act ivity on SDS PAGE containing casein One of the ECPases from the su ernatant w as urified by ammonium sulfate reci it ation, anion-exchange chro- matogra hy on DEAE-cellulose and Se hadex G-200 gel filt ration chromatogra hy . SDSof the u- rified rotease indicated that 2-ME reduced sam le migrated at 35 000, w hile non-reduced sam le migrated at 54 000. T his rot ease was accordingly designat ed ECPase 54. The activit y of ECPase 54 w as inhibited , 56 30 . 54 by EDTA but not by PMSF or by heating at for mi