Thomas E L
J Bacteriol. 1984 Jan;157(1):240-6. doi: 10.1128/jb.157.1.240-246.1984.
Incubation of Streptococcus mutans cells with certain disulfide compounds resulted in accumulation of reduced sulfhydryl compounds in the extracellular medium or in both the medium and the cells. Oxidized lipoic acid and lipoamide competed for reduction. At high concentrations, these compounds were reduced at rates comparable to that of glucose metabolism, and all of the increase in sulfhydryls was in the medium. Cystamine did not compete with these compounds for reduction but was also reduced at high rates and low apparent affinity, and all of the cysteamine produced from cystamine accumulated in the medium. In contrast, glutathione disulfide (GSSG) and L-cystine were reduced slowly but with high apparent affinity, and 60 to 80% of the increase in sulfhydryls was intracellular. NADH-dependent lipoic acid or lipoamide reductase activity was present in the particulate (wall-plus-membrane) fraction, whereas NADPH-dependent GSSG reductase activity was present in the soluble (cytoplasmic) fraction. Two transport systems for disulfide and sulfhydryl compounds were distinguished. GSSG, L-cystine, and reduced glutathione competed for uptake. L-Cysteine was taken up by a separate system that also accepted L-penicillamine and D-cysteine as substrates. Uptake of glutathione or L-cysteine, or the uptake and reduction of GSSG or L-cystine, resulted in up to a 10-fold increase in cell sulfhydryl content that raised intracellular concentrations to between 30 and 40 mM. These reductase and transport systems enable S. mutans cells to create a reducing environment in both the extracellular medium and the cytoplasm.
用某些二硫化合物培养变形链球菌细胞,导致细胞外培养基或培养基与细胞中均积累了还原型巯基化合物。氧化型硫辛酸和硫辛酰胺竞争还原作用。在高浓度下,这些化合物的还原速率与葡萄糖代谢的速率相当,并且所有巯基的增加都在培养基中。胱胺不与这些化合物竞争还原作用,但也能以高还原速率和低表观亲和力被还原,并且由胱胺产生的所有半胱胺都积累在培养基中。相比之下,谷胱甘肽二硫化物(GSSG)和L-胱氨酸还原缓慢但具有高表观亲和力,并且60%至80%的巯基增加是在细胞内。NADH依赖的硫辛酸或硫辛酰胺还原酶活性存在于颗粒(细胞壁加细胞膜)部分,而NADPH依赖的GSSG还原酶活性存在于可溶性(细胞质)部分。区分了两种二硫化合物和巯基化合物的转运系统。GSSG、L-胱氨酸和还原型谷胱甘肽竞争摄取。L-半胱氨酸通过一个单独的系统摄取,该系统也接受L-青霉胺和D-半胱氨酸作为底物。摄取谷胱甘肽或L-半胱氨酸,或摄取并还原GSSG或L-胱氨酸,会使细胞巯基含量增加多达10倍,从而将细胞内浓度提高到30至40 mM之间。这些还原酶和转运系统使变形链球菌细胞能够在细胞外培养基和细胞质中创造一个还原环境。