The hyperthermophilic archaeon Thermococcus kodakarensis can grow on pyruvate or maltooligosaccharides through H₂ fermentation. H₂ production levels of members of the Thermococcales are high, and studies to improve their production potential have been reported. Although H₂ production is primary metabolism, here we aimed to partially uncouple cell growth and H₂ production of T. kodakarensis. Additional A₁-type ATPase genes were introduced into T. kodakarensis KU216 under the control of two promoters; the strong constitutive cell surface glycoprotein promoter, P_csg, and the sugar-inducible fructose-1,6-bisphosphate aldolase promoter, P_fba. Whereas cells with the A₁-type ATPase genes under the control of P_csg displayed only trace levels of growth, cells with P_fba (strain KUA-PF) displayed growth sufficient for further analysis. Increased levels of A₁-type ATPase protein were detected in KUA-PF cells grown on pyruvate or maltodextrin, when compared to the levels in the host strain KU216. The growth and H₂ production levels of strain KUA-PF with pyruvate or maltodextrin as a carbon and electron source were analyzed and compared to those of the host strain KU216. Compared to a small decrease in total H₂ production, significantly larger decreases in cell growth were observed, resulting in an increase in cell-specific H₂ production. Quantification of the substrate also revealed that ATPase overexpression led to increased cell-specific pyruvate and maltodextrin consumptions. The results clearly indicate that ATPase production results in partial uncoupling of cell growth and H₂ production in T. kodakarensis.

嗜高温古生菌Thercococcus kodakarensis可以通过H ₂发酵在丙酮酸或麦芽低聚糖上生长。嗜热球菌成员的H ₂产生水平很高，并且已经报道了提高其产生潜力的研究。尽管H _2的产生是主要的新陈代谢，但在这里我们的目标是部分解耦柯达卡氏菌的细胞生长和H ₂产生。在两个启动子的控制下，将另外的一个A ₁型ATPase基因导入了柯达卡氏菌KU216。强本构细胞表面糖蛋白启动子P _csg，和糖诱导的果糖-1,6-双磷酸醛缩酶启动子P _fba。在P _csg的控制下，具有A ₁型ATPase基因的细胞仅_表现出痕量的生长，而具有P _fba（菌株KUA-PF）的细胞表现出足以进行进一步分析的生长。当与宿主菌株KU216中的水平相比时，在丙酮酸或麦芽糖糊精上生长的KUA-PF细胞中检测到A ₁型ATP酶蛋白水平增加。分析了丙酮酸或麦芽糊精作为碳源和电子源的菌株KUA-PF的生长和H ₂产生水平，并与宿主菌株KU216进行了比较。与总H ₂的少量减少相比产生细胞时，观察到细胞生长明显减少，导致细胞特异性H ₂产生增加。底物的定量还揭示了ATP酶的过表达导致细胞特异性丙酮酸和麦芽糊精的消耗增加。结果清楚地表明，ATP酶的产生导致了T. kodakarensis的细胞生长和H ₂产生的部分解偶联。