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Physicochemical Fractionation of Extracellular Cornea-Damaging Proteases of Pseudomonas aeruginosa

机译：铜绿假单胞菌胞外角膜损伤蛋白酶的理化分级

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Fractionation of the culture supernatant fluids of a cornea-virulent strain of Pseudomonas aeruginosa by ammonium sulfate precipitation, diafiltration, isoelectric focusing, ion-exchange chromatography, gel filtration, and sucrose density gradient centrifugation failed to separate the rabbit cornea-damaging activity and the in vitro protease activity of the preparations. Three proteases having similar molecular weights (approximately 20,000) and isoelectric points of approximately 4.6, 5.8, and 8.8 were obtained free of detectable amounts of other known extracellular pseudomonal enzymes. Heating a mixture of the three proteases for 15 min at 80 C resulted in complete loss of protease and cornea-damaging activities. The sterile culture filtrate of a nonproteolytic but lethal toxin-producing strain of P. aeruginosa did not contain cornea-damaging activity. Cultivation of the proteolytic strain in broth containing 4.7% ammonium sulfate yielded a culture supernatant fluid free of protease and cornea-damaging activities. The results obtained support the conclusion that a cornea-virulent strain of P. aeruginosa can produce, in vitro, at least three different extracellular proteases capable of eliciting rapid and extensive damage to rabbit corneas.
机译：通过硫酸铵沉淀，渗滤，等电聚焦，离子交换色谱，凝胶过滤和蔗糖密度梯度离心分离角膜致病性铜绿假单胞菌菌株的培养物上清液无法分离兔制剂的角膜破坏活性和体外蛋白酶活性。获得了三种具有相似分子量（约20,000）和等电点分别约为4.6、5.8和8.8的蛋白酶，其中没有可检测量的其他已知细胞外假单胞菌酶。将这三种蛋白酶的混合物在80°C下加热15分钟，会导致蛋白酶的完全丧失和破坏角膜的活性。非蛋白水解但致死毒素的 P菌株的无菌培养滤液。铜绿假单胞菌不具有破坏角膜的活性。在含有4.7％硫酸铵的肉汤中蛋白水解菌株的培养产生了不含蛋白酶和角膜破坏活性的培养上清液。获得的结果支持了 P的角膜毒力菌株的结论。铜绿假单胞菌可以在体外产生至少三种不同的细胞外蛋白酶，能够引起对兔子角膜的快速而广泛的损伤。

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