Division of Life Science, Korea Basic Science Institute, Daejeon 305-333, Republic of Korea.
J Proteomics. 2011 Sep 6;74(10):1926-33. doi: 10.1016/j.jprot.2011.05.010. Epub 2011 May 15.
Thermococcus onnurineus NA1 is a typical sulfur-reducing hyperthermophilic archaeon. Genome sequence analysis has shown that T. onnurineus NA1 retains the metabolic pathways necessary not only for organotrophic, but also for carboxydotrophic, growth. T. onnurineus NA1 carboxydotrophic growth may result in hydrogen production, as this archaeon produces hydrogen during oxidizing formate. In this study, we profiled the proteome of T. onnurineus NA1 cultured under carboxydotrophic conditions using CO as an electron donor by the SDS-PAGE/LC-MS/MS method. A total of 1395 proteins were identified by two independent proteomic analyses, which corresponds to ~71% of the total predicted open reading frames. To our knowledge, this level of identification coverage exceeds those of other global proteome profiling studies in Archaea. Furthermore, the biological functions of the identified proteins were predicted and cognate enzymes were mapped to the appropriate metabolic pathways. More than 90% of the genes belonging to hydrogenase gene clusters such as Mbx, Sulf-I, Mbh, Hyg4-I, Hyg4-II, and Hyf4-III were expressed during CO culture. This means that hydrogenases induced under carboxydotrophic conditions surpass those induced under an organotrophic condition (yeast extract-peptone-sulfur). Our data suggest that hydrogen production is caused by the expression and functional assembly of T. onnurineus NA1 hydrogenase genes during culture in the presence of CO.
火球菌（Thermococcus onnurineus）NA1 是一种典型的硫还原高温古菌。基因组序列分析表明，T. onnurineus NA1 保留了不仅用于有机营养生长，而且用于碳固定生长所必需的代谢途径。T. onnurineus NA1 的碳固定生长可能导致氢气的产生，因为这种古菌在氧化甲酸盐时会产生氢气。在这项研究中，我们使用 SDS-PAGE/LC-MS/MS 方法，通过以 CO 作为电子供体对培养在碳固定条件下的 T. onnurineus NA1 的蛋白质组进行了分析。通过两次独立的蛋白质组学分析鉴定了 1395 种蛋白质，这对应于总预测开放阅读框的约 71%。据我们所知，这种鉴定覆盖率超过了其他古菌的全球蛋白质组分析研究。此外，还预测了鉴定出的蛋白质的生物学功能，并将同源酶映射到适当的代谢途径上。属于氢化酶基因簇（如 Mbx、Sulf-I、Mbh、Hyg4-I、Hyg4-II 和 Hyf4-III）的 90%以上的基因在 CO 培养期间表达。这意味着在碳固定条件下诱导的氢化酶超过了在有机营养条件（酵母提取物-蛋白胨-硫）下诱导的氢化酶。我们的数据表明，在 CO 存在的条件下培养时，T. onnurineus NA1 氢化酶基因的表达和功能组装导致了氢气的产生。
