Kiyotani K, Tasaka H, Matsuo Y
Microbiol Immunol. 1983;27(8):695-708. doi: 10.1111/j.1348-0421.1983.tb00632.x.
Factors inhibitory to beta-glucuronidase were found in the culture filtrate and in a bacillary extract of Mycobacterium tuberculosis H37Rv grown for 6 weeks on Sauton medium. The inhibitors were purified by ammonium sulfate fractionation, treatment with n-butanol and streptomycin, and chromatography on DEAE-Sepharose CL-6B. Two inhibitors were obtained from the culture filtrate. The molecular weights were estimated to be 25,500 and 15,500 by gel filtration on a Sephadex G-75 column. Three inhibitors were purified from the bacillary extract, two of which were similar to those from the culture filtrate. The molecular weight of the third inhibitor was 21,000. However, the molecular weight of all the denatured inhibitors was 8,600 in the presence of sodium dodecyl sulfate. The inhibitors contained extremely high amounts of glutamic and aspartic acids and had a highly acidic isoelectric point of pH 2.5. The inhibitors acted noncompetitively against beta-glucuronidase of guinea pig origin at an optimal pH 4.5. beta-Glucuronidases from human peripheral leukocytes and beef liver were partially sensitive to the inhibitors; all the other enzymes tested for sensitivity were unaffected by the inhibitors.
在结核分枝杆菌H37Rv在索顿培养基上培养6周后的培养滤液和杆菌提取物中发现了β-葡萄糖醛酸酶的抑制因子。通过硫酸铵分级分离、正丁醇和链霉素处理以及DEAE-琼脂糖CL-6B柱色谱法对抑制剂进行了纯化。从培养滤液中获得了两种抑制剂。通过在Sephadex G-75柱上进行凝胶过滤，估计其分子量分别为25,500和15,500。从杆菌提取物中纯化出三种抑制剂，其中两种与培养滤液中的相似。第三种抑制剂的分子量为21,000。然而，在十二烷基硫酸钠存在下，所有变性抑制剂的分子量均为8,600。这些抑制剂含有极大量的谷氨酸和天冬氨酸，等电点极低，为pH 2.5。在最佳pH 4.5时，这些抑制剂对豚鼠来源的β-葡萄糖醛酸酶起非竞争性抑制作用。人外周血白细胞和牛肝中的β-葡萄糖醛酸酶对这些抑制剂部分敏感；所测试的其他所有酶的敏感性均不受这些抑制剂的影响。
