GENETIC AND BIOCHEMICAL DIVERSITY OF UREASES OF PROTEUS, PROVIDENCIA, AND MORGANELLA SPECIES ISOLATED FROM URINARY-TRACT INFECTION
GENETIC AND BIOCHEMICAL DIVERSITY OF UREASES OF PROTEUS, PROVIDENCIA, AND MORGANELLA SPECIES ISOLATED FROM URINARY-TRACT INFECTION
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DOI：
10.1128/iai.55.9.2198-2203.1987
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发表时间：
1987-09-01
影响因子：
3.1
通讯作者：
MOBLEY, HLT
中科院分区：
文献类型：
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作者：
JONES, BD;MOBLEY, HLT
细菌性尿素酶，特别是来自奇异变形杆菌的细菌性尿素酶，已被认为是泌尿和肾结石、梗阻或导尿管以及肾盂肾炎形成的促成因素。来自居住在两个慢性病护理机构的32名患者的每周尿液样本（n = 1，135），其中导尿管放置在适当位置以用于≥30天产生5，088个表型和血清型多样性的细菌分离株，105 CFU/ml。总共86%的标本含有至少一种尿素酶阳性菌种，3，939株革兰氏阴性杆菌中有46%为尿素酶阳性。为了研究尿素酶决定簇的遗传相关性，将来自斯氏普罗威登斯菌、雷氏普罗威登斯菌、奇异变形杆菌、普通变形杆菌和摩氏摩根菌的各50个尿素酶阳性分离株的全细胞DNA与来自斯氏普罗威登斯菌BE 2467的尿素酶操纵子内的尿素酶基因探针杂交。结果表明，斯氏普罗威登斯菌98株，雷氏普罗威登斯菌100株，奇异变形杆菌70株，M.摩根菌为0，普通菌为0。电泳迁移率的尿素酶从代表性的分离株揭示了九个不同的模式之间的五个物种。尿素酶基因探针与除M.摩根尼。不同物种之间的片段大小不同。通过Sephacryl S-300色谱法测定的酶的分子大小为280千道尔顿（kDa）（奇异假单胞菌）、323至337 kDa（斯氏普罗威登斯菌、雷氏普罗威登斯菌、奇异假单胞菌、普通普罗威登斯菌）、620 kDa（雷氏普罗威登斯菌）和> 700 kDa（M. morganii，Providencia rettgeri）。Km范围为0.7 mM尿素（M）。对于奇异变形杆菌分离物，将morganii菌株添加至60 mM尿素。一般来说，奇异变形杆菌尿素酶表现出对底物的亲和力较低，但水解尿素的速率比其他物种的酶快6- 25倍，这可能解释了该物种与结石形成的频繁关联。
Bacterial urease, particularly from Proteus mirabilis, has been implicated as a contribution factor in the formation of urinary and kidney stones, obstruction or urinary catheters, and pyelonephritis. Weekly urine specimens (n = 1,135) from 32 patients, residing at two chronic-care facilities, with urinary catheters in place for .gtoreq. 30 days yielded 5,088 phenotypically and serotypically diverse bacterial isolates at .gtoreq. 105 CFU/ml. A total of 86% of specimens contained at least one urease-positive species, and 46% of 3,939 gram-negative bacilli were urease positive. For investigation of genetic relatedness of urease determinant, whole-cell DNA from 50 urease-positive isolates each of Providencia stuartii, Providencia rettgeri, P. mirabilis, Proteus vulgaris, and Morganella morganii were hybridized with a urease gene probe derived from within the urease operon of Providencia stuartii BE2467. The percentage of strains hybridizing with the gene probe was 98 for Providencia stuartii, 100 for Providencia rettgeri, 70 for P. mirabilis, 2 for M. morganii, and 0 for P. vulgaris. Electrophoretic mobilities of ureases from representative isolates revealed nine different patterns among the five species. The urease gene probe hybridized with fragments of HindIII-digested chromosomal DNA from all isolates except M. morganii. Fragment sizes differed between species. Molecular sizes of the enzymes, determined by Sephacryl S-300 chromatography, were found to be 280 kilodaltons (kDa) (P. mirabilis), 323 to 337 kDa (Providencia stuartii, Providencia rettgeri, P. mirabilis, P. vulgaris), 620 kDa (Providencia rettgeri), and > 700 kDa (M. morganii, Providencia rettgeri). Kms ranged from 0.7 mM urea for M. morganii to 60 mM urea for a P. mirabilis isolate. In general, P. mirabilis ureases demonstrated lower affinities for substrate but hydrolyzed urea at rates 6- to 25-fold faster than did enzymes from other species, which may explain the frequent association of this species with stone formation.