Laboratory of Biodefense & Regulation, Osaka University of Pharmaceutical Sciences, Nasahara, Takatsuki, Osaka 569–1094, Japan.
Microbes Environ. 2013;28(2):180-6. doi: 10.1264/jsme2.me12174. Epub 2013 Apr 17.
An environmental isolate of Salmonella Enteritidis (SE), grown to the logarithmic phase, rapidly lost culturability by the addition of 3 mM H2O2 to cultures grown in Luria-Bertani (LB) medium; however, some H2O2-treated bacteria regained their culturability in M9 minimal medium, if sodium pyruvate was present at at least 0.3 mM. In addition, most pyruvate analogues, such as bromopyruvate or phenylpyruvate, did not show restoration activity similar to that of pyruvate, except in the case of α-ketobutyrate. Further analysis of the mechanism underlying the resuscitation by pyruvate revealed that although many of the bacteria showed respiratory activity on CTC (5-cyano-2,3-di-(p-tolyl) tetrazolium chloride) reduction with or without pyruvate, the biosynthesis of DNA and protein synthesis were quite different in the presence or absence of pyruvate, i.e., pyruvate endowed the cells with the ability to incorporate much more radio-label into precursors during the resuscitation process. These results suggest that pyruvate is one of the key molecules working in the resuscitation process by taking bacteria from the non-culturable state to the growing and colony-forming state by triggering the synthesis of macromolecules such as DNA and protein.
一株环境分离的肠炎沙门氏菌（SE）在对数生长期时，用 3mM H2O2 处理在 LB 培养基中生长的细菌，其可培养性迅速丧失；然而，如果在 M9 最低培养基中存在至少 0.3mM 的丙酮酸，那么一些经 H2O2 处理的细菌可以恢复其可培养性。此外，大多数丙酮酸类似物，如溴丙酮酸或苯丙酮酸，除了α-酮丁酸外，都没有表现出类似于丙酮酸的恢复活性。对丙酮酸复苏机制的进一步分析表明，尽管许多细菌在用或不用丙酮酸的情况下均可进行 CTC（5-氰基-2,3-二（对甲苯基）四唑氯）还原的呼吸活性，但在有或没有丙酮酸的情况下，DNA 的生物合成和蛋白质的合成大不相同，即丙酮酸赋予细胞在复苏过程中整合更多放射性标记物到前体中的能力。这些结果表明，丙酮酸是使细菌从不可培养状态到生长和形成菌落状态复苏的关键分子之一，通过触发 DNA 和蛋白质等大分子的合成来实现。
