Wada Keisuke, Toya Yoshihiro, Matsuda Fumio, Shimizu Hiroshi
Department of Bioinformatic Engineering, Graduate School of Information Science and Technology, Osaka University, 1-5 Yamadaoka, Suita, Osaka, 565-0871, Japan.
Research Institute for Sustainable Chemistry, National Institute of Industrial Advanced Science and Technology (AIST), Tsukuba Central 5-2, 1-1-1 Higashi, Tsukuba, Ibaraki, 305-8565, Japan.
Appl Biochem Biotechnol. 2025 May;197(5):2944-2957. doi: 10.1007/s12010-024-05138-4. Epub 2025 Jan 15.
Cyanobacteria are advantageous hosts for industrial applications toward achieving sustainable society due to their unique and superior properties such as atmospheric CO fixation via photosynthesis. However, cyanobacterial productivities tend to be weak compared to heterotrophic microbes. To enhance them, it is necessary to understand the fundamental metabolic mechanisms unique to cyanobacteria. In cyanobacteria, NADPH and ATP regenerated by linear and cyclic electron transfers using light energy are consumed by CO fixation in a central metabolic pathway. The previous study demonstrated that the strain deleted a part of respiratory chain complex (ΔndhF1) perturbed NADPH levels and photosynthetic activity in Synechocystis sp. PCC 6803. It is expected that disruption of ndhF1 would result in a decrease in the function of cyclic electron transfer, which controls the ATP/NAD(P)H production ratio properly. In this study, we evaluated the effects of ndhF1 deletion on central metabolism and photosynthesis by C-metabolic flux analysis. As results of culturing the control and ΔndhF1 strains in a medium containing [1,2-C] glucose and estimating the flux distribution, CO fixation rate by RuBisCO was decreased to be less than half in the ΔndhF1 strain. In addition, the regeneration rate of NAD(P)H and ATP by the photosystem, which can be estimated from the flux distribution, also decreased to be less than half in the ΔndhF1 strain, whereas no significant difference was observed in ATP/NAD(P)H production ratio between the control and the ΔndhF1 strains. Our result suggests that the ratio of utilization of cyclic electron transfer is not reduced in the ΔndhF1 strain unexpectedly.
由于蓝藻具有独特且优越的特性,如通过光合作用固定大气中的二氧化碳,因此在实现可持续社会的工业应用中是理想的宿主。然而,与异养微生物相比,蓝藻的生产力往往较弱。为了提高其生产力,有必要了解蓝藻特有的基本代谢机制。在蓝藻中,利用光能通过线性和循环电子传递再生的NADPH和ATP在中心代谢途径中被二氧化碳固定所消耗。先前的研究表明,缺失部分呼吸链复合物(ΔndhF1)的菌株会扰乱集胞藻PCC 6803中的NADPH水平和光合活性。预计ndhF1的缺失会导致循环电子传递功能下降,而循环电子传递可适当控制ATP/NAD(P)H的产生比率。在本研究中我们通过C代谢通量分析评估了ndhF1缺失对中心代谢和光合作用的影响。在含有[1,2-C]葡萄糖的培养基中培养对照菌株和ΔndhF1菌株并估算通量分布,结果显示,在ΔndhF1菌株中,RuBisCO的二氧化碳固定率降至不到对照菌株的一半。此外,从通量分布估算得出,光合系统再生NAD(P)H和ATP的速率在ΔndhF1菌株中也降至不到对照菌株的一半,而对照菌株和ΔndhF1菌株之间的ATP/NAD(P)H产生比率未观察到显著差异。我们的结果表明,出乎意料的是,在ΔndhF1菌株中循环电子传递的利用比率并未降低。