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首页> 外文期刊>Infection and immunity >Mechanism of the Adherence of Streptococcus mutans to Smooth Surfaces III. Purification and Properties of the Enzyme Complex Responsible for Adherence
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Mechanism of the Adherence of Streptococcus mutans to Smooth Surfaces III. Purification and Properties of the Enzyme Complex Responsible for Adherence

机译：变形链球菌粘附于光滑表面的机理III。负责粘附的酶复合物的纯化和性质

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Enzymes which possess the ability to cause the adherence of Streptococcus mutans cells to a smooth glass surface were purified 1,100 times by chromatography on agarose gel followed by hydroxylapatite gel. During the purification procedures, the enzymes from strain HS6 (group a) were examined for the synthesis of water-soluble and water-insoluble polysaccharide and the ability to produce adherence. The enzyme preparations producing adherence of the S. mutans cells in the presence of sucrose possessed a molecular size of about 400,000 to 2,000,000 and were composed of approximately equivalent amounts of dextran and levan sucrases and 5 to 30% polysaccharide. The most highly purified preparation contained a negligible amount of contaminating protein as judged by polyacrylamide gel electrophoresis, immunoelectrophoresis, and gel diffusion. In these three tests, the location of the enzyme responsible for the synthesis of insoluble polymer was detected by embedding or covering the enzyme-containing gel with a layer of sucrose-containing agarose gel and observing the formation of insoluble polymer. During purification the ability of all fractions to produce adherence was parallel with the enzyme activity responsible for the synthesis of insoluble polysaccharide from sucrose. About two-thirds of the sucrase enzyme complex in the S. mutans culture fluid synthesized water-soluble polymer. This complex, obtained by filtration through agarose gel, was smaller in molecular size, lower in sugar content, and did not produce adherence, in contrast to the enzyme complex which possessed adherence activity. The inhibition of the enzyme complex synthesizing soluble polymer required more anti-synthetase serum than that required to inhibit the synthesis of water-insoluble polymer. It is not known whether the lack of adherence activity in this enzyme was due to its smaller size and lower sugar content or the absence of unknown factors which are essential for its activity. The carbohydrate in these enzyme preparations, composed of glucose, may represent a primer molecule and/or a remnant of the polymer synthesized by the enzyme. The enzyme activity was not inhibited by anti-dextran globulin.
机译：通过在琼脂糖凝胶上层析，然后在羟磷灰石凝胶上层析，将具有导致变形链球菌细胞粘附到光滑玻璃表面的能力的酶纯化1,100倍。在纯化过程中，检查了HS6菌株（ a 组）中的酶的水溶性和非水溶性多糖的合成以及产生粘附的能力。产生 S粘附力的酶制剂。存在蔗糖的变形细胞具有大约400,000至2,000,000的分子大小，并由大约当量的右旋糖酐和左旋蔗糖糖和5％至30％的多糖组成。通过聚丙烯酰胺凝胶电泳，免疫电泳和凝胶扩散判断，纯化程度最高的制剂所含污染蛋白的量可忽略不计。在这三个测试中，通过将含酶的凝胶包埋或覆盖一层含蔗糖的琼脂糖凝胶，并观察不溶性聚合物的形成，来检测负责不溶性聚合物合成的酶的位置。在纯化过程中，所有级分产生粘附的能力与负责由蔗糖合成不溶多糖的酶活性平行。 S中约三分之二的蔗糖酶复合物。变形菌培养液合成水溶性高分子。与具有粘附活性的酶复合物相比，通过琼脂糖凝胶过滤获得的该复合物分子尺寸较小，糖含量较低，并且不产生粘附。与抑制水不溶性聚合物的合成相比，抑制酶复合物合成可溶的聚合物需要更多的抗合成酶血清。尚不清楚该酶缺乏粘附活性是由于其较小的尺寸和较低的糖含量还是由于缺乏对其活性至关重要的未知因素所致。这些由葡萄糖组成的酶制剂中的碳水化合物可以代表引物分子和/或由该酶合成的聚合物的残余物。酶活性不受抗葡聚糖球蛋白的抑制。

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