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鸟分枝杆菌、胞内分枝杆菌和瘰疬分枝杆菌的芳基硫酸酯酶活性。

Arylsulfatase activity of Mycobacterium avium, M. intracellulare, and M. scrofulaceum.

作者信息

Falkinham J O

机构信息

Department of Biology, Virginia Polytechnic Institute and State University, Blacksburg 24061.

出版信息

Int J Syst Bacteriol. 1990 Jan;40(1):66-70. doi: 10.1099/00207713-40-1-66.

Abstract

A rapid (3-h) arylsulfatase assay for cell suspensions of mycobacteria, in which p-nitrophenyl sulfate is used as the substrate, was developed. Arylsulfatase activity was found in cell suspensions of representative strains of Mycobacterium avium, Mycobacterium intracellulare, and Mycobacterium scrofulaceum grown without the substrate in either Middlebrook 7H9 medium containing 0.2% (wt/vol) glucose and 0.05% (vol/vol) Tween 80 or Dubos broth medium, but was absent in cells grown in a low-pH, minimal medium containing 1% (vol/vol) Tween 80 as the sole carbon source. The levels of arylsulfatase activity of representatives of all three species were equal whether the activity was measured at pH 5.5, 6.5, or 7.5 and whether the cells were suspended in phosphate or Tris buffer. The addition of high levels of sulfate (present in the low-pH, Tween 80-containing medium) to Middlebrook 7H9 medium resulted in significantly lower levels of arylsulfatase activity in strains of M. scrofulaceum, but did not affect the levels in either M. avium or M. intracellulare. The levels of arylsulfatase activity were highest in M. avium, intermediate in M. intracellulare, and lowest in M. scrofulaceum strains. Polyacrylamide gel electrophoresis of crude extracts from late-log-phase cells of representatives of each species produced activity bands of unique mobility (one in M. avium, three in M. intracellulare [82, 5, and 13%], and two in M. scrofulaceum [60 and 40%]).

摘要

开发了一种快速(3小时)的分枝杆菌细胞悬液芳基硫酸酯酶测定法,该方法以对硝基苯硫酸酯作为底物。在不含底物的情况下,于含有0.2%(重量/体积)葡萄糖和0.05%(体积/体积)吐温80的Middlebrook 7H9培养基或Dubos肉汤培养基中培养的鸟分枝杆菌、胞内分枝杆菌和瘰疬分枝杆菌代表性菌株的细胞悬液中发现了芳基硫酸酯酶活性,但在以1%(体积/体积)吐温80作为唯一碳源的低pH、基本培养基中生长的细胞中未发现该活性。无论在pH 5.5、6.5还是7.5下测量活性,也无论细胞悬浮在磷酸盐缓冲液还是Tris缓冲液中,所有三个菌种的代表性菌株的芳基硫酸酯酶活性水平均相等。向Middlebrook 7H9培养基中添加高水平的硫酸盐(存在于含吐温80的低pH培养基中)会导致瘰疬分枝杆菌菌株中的芳基硫酸酯酶活性水平显著降低,但不影响鸟分枝杆菌或胞内分枝杆菌中的活性水平。芳基硫酸酯酶活性水平在鸟分枝杆菌中最高,在胞内分枝杆菌中居中,在瘰疬分枝杆菌菌株中最低。对每个菌种代表性菌株对数生长后期细胞的粗提物进行聚丙烯酰胺凝胶电泳,产生了具有独特迁移率的活性条带(鸟分枝杆菌中有一条,胞内分枝杆菌中有三条[82%、5%和13%],瘰疬分枝杆菌中有两条[60%和40%])。

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