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首页> 外文期刊>Archives of microbiology >The Serratia marcescens hemolysin is secreted but not activated by stable protoplast-type L-forms of Proteus mirabilis
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The Serratia marcescens hemolysin is secreted but not activated by stable protoplast-type L-forms of Proteus mirabilis

机译：粘质沙雷氏菌的溶血素被稳定的原生质体型L型变形杆菌分泌但未被激活

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摘要

The outer-membrane protein ShlB of Serratia marcescens activates and secretes hemolytic ShlA into the culture medium. Without ShlB, inactive ShlA (termed ShlA*) remains in the periplasm. Since Proteus mirabilis L-form cells lack an outer membrane and a periplasm, it was of interest to determine in which compartment recombinant ShlA* and ShlB are localized and whether ShlB activates ShlA*. The cloned shlB and shlA genes were transcribed in P. mirabilis stable L-form cells by the temperature-inducible phage T7 RNA polymerase. Radiolabeling, Western blotting, and complementation with C-terminally truncated ShlA (ShlA255) identified inactive ShlA* in the culture supernatant. ShlB remained cell-bound and did not activate ShlA without integration in an outer membrane. Although hemolytic ShlA added to L-form cells had access to the cytoplasmic membrane, it did not affect L-form cells. Synthesis of the large ShlA protein (165 kDa) in P. mirabilis L-form cells under phage T7 promoter control demonstrates that L-form cells are suitable for the synthesis and secretion of large recombinant proteins. This property and the easy isolation of released proteins make L-form cells suitable for the biotechnological production of proteins. [References: 27]
机译：粘质沙雷氏菌的外膜蛋白ShlB激活并将溶血性ShlA分泌到培养基中。没有ShlB，无活性的ShlA（称为ShlA *）保留在周质中。由于奇异变形杆菌L型细胞缺少外膜和周质，因此有兴趣确定重组ShlA *和ShlB位于哪个隔室中以及ShlB是否激活ShlA *。通过温度诱导型噬菌体T7 RNA聚合酶将克隆的shlB和shlA基因转录到奇异毕赤酵母稳定的L型细胞中。放射标记，蛋白质印迹和与C末端截短的ShlA（ShlA255）互补，在培养上清液中鉴定出无活性的ShlA *。如果没有整合在外膜中，ShlB仍然与细胞结合，并且不会激活ShlA。尽管添加到L型细胞的溶血性ShlA可以进入细胞质膜，但它不会影响L型细胞。在噬菌体T7启动子控制下，在奇异毕赤酵母L型细胞中合成大型ShlA蛋白（165 kDa）表明，L型细胞适合于大型重组蛋白的合成和分泌。这种特性和释放蛋白质的容易分离使L型细胞适合于蛋白质的生物技术生产。 [参考：27]

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