PURIFICATION AND CHARACTERIZATION OF A HEMOLYSIN PRODUCED BY A CLINICAL ISOLATE OF KANAGAWA PHENOMENON-NEGATIVE VIBRIO-PARAHAEMOLYTICUS AND RELATED TO THE THERMOSTABLE DIRECT HEMOLYSIN
PURIFICATION AND CHARACTERIZATION OF A HEMOLYSIN PRODUCED BY A CLINICAL ISOLATE OF KANAGAWA PHENOMENON-NEGATIVE VIBRIO-PARAHAEMOLYTICUS AND RELATED TO THE THERMOSTABLE DIRECT HEMOLYSIN
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DOI：
10.1128/iai.56.4.961-965.1988
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发表时间：
1988-04-01
影响因子：
3.1
通讯作者：
MIWATANI, T
中科院分区：
文献类型：
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作者：
HONDA, T;NI, YX;MIWATANI, T
对一株临床分离的神奈川现象阴性副溶血弧菌（4037株）进行了研究。虽然通过各种常规试验判定该菌株为神奈川现象阴性，但它产生一种新的溶血素（命名为Vp-TRH，为耐热直接溶血素[Vp-TDH]相关溶血素），该溶血素与普通神奈川现象阳性副溶血弧菌产生的Vp-TDH相关。Vp-TRH经硫酸铵分级分离和连续的DEAE-纤维素、羟基磷灰石和Mono Q柱层析纯化。Sephadex G-100凝胶过滤法测得Vp-TRH的分子量为48，000，十二烷基硫酸钠平板凝胶电泳法测得Vp-TRH亚基的分子量为23，000。因此，与Vp-TDH一样，Vp-TRH似乎也由两个亚基组成。测定Vp-TRH的等电点为4.6。Vp-TRH对各种动物红细胞的裂解活性与Vp-TDH不同，尤其是小牛、鸡和绵羊红细胞。Vp-TRH的溶血活性在60 ℃热处理时不稳定。与Vp-TDH不同，通过Ouchterlony、中和和乳胶凝集试验证明了Vp-TRH和Vp-TDH的免疫学相似性，但不具有同一性。因此，我们得出结论，这种神奈川现象阴性的副溶血性弧菌的临床分离株产生一种新型溶血素，这种溶血素与通常由神奈川现象阳性的副溶血性弧菌产生的Vp-TDH相似，但不完全相同。
A clinical isolate (strain 4037) of Kanagawa phenomenon-negative Vibrio parahaemoyticus was studied. Although the strain was judged to be Kanagawa phenomenon-negative by various conventional tests, it produced a new hemolysin (named Vp-TRH, for thermostable direct hemolysin [Vp-TDH]-related hemolysin) that was related to the Vp-TDH produced by ordinary Kanagawa phenomenon-positive V. parahaemolyticus. Vp-TRH was purified by ammonium sulfate fractionation and successive column chromatographies on DEAE-cellulose, hydroxyapatite, and Mono Q. The molecular weight of Vp-TRH was estimated as 48,000 by Sephadex G-100 gel filtration, and the molecular weight of the subunit was estimated to be 23,000 by sodium dodecyl sulfate-slab gel electrophoresis. Thus, like Vp-TDH, Vp-TRH seems to be composed of two subunits. The isoelectric point of Vp-TRH was determined to be 4.6. Vp-TRH showed lytic activities different from those of Vp-TDH on erythrocytes from various animals, especially those from calves, chickens, and sheep. The hemolytic activity of Vp-TRH was labile on heat treatment at 60.degree. C for 10 min, unlike that of Vp-TDH. The immunological similarities, but not the identities of Vp-TRH and Vp-TDH, were demonstrated by Ouchterlony, neutralization, and latex agglutination tests. Thus, we conclude that this clinical isolate of Kanagawa phenomenon-negative V. parahaemolyticus produces a new type of hemolysin that is similar, but not identical, to Vp-TDH, which is usually produced by Kanagawa phenomenon-positive V. parahaemolyticus.