Graduate School of Science and Engineering, Saitama University, 225 Shimo-Okubo, Sakura-ku, Saitama-city, Saitama, 338-8570, Japan.
Graduate School of Science, Ritsumeikan University, Kusatsu, Japan.
Plant J. 2019 May;98(4):654-666. doi: 10.1111/tpj.14262. Epub 2019 Mar 14.
Pyridine nucleotides (NAD(P)(H)) are electron carriers that are the driving forces in various metabolic pathways. Phosphorylation of NAD(H) to NADP(H) is performed by the enzyme NAD kinase (NADK). Synechocystis sp. PCC 6803 harbors two genes (sll1415 and slr0400) that encode proteins with NADK homology. When genetic mutants for sll1415 and slr0400 (Δ1415 and Δ0400, respectively) were cultured under photoheterotrophic growth conditions only the Δ1415 cells showed a growth defect. In wild-type cells, the sll1415 transcript accumulated after the cells were transferred to photoheterotrophic conditions. Furthermore, NAD(P)(H) measurements demonstrated that a dynamic metabolic conversion was implemented during the adaptation from photoautotrophic to photoheterotrophic conditions. Electron microscopy observation and biochemistry quantification demonstrated the accumulation of glycogen in the Δ1415 cells under photoheterotrophic conditions at 96 h. Quantitative real-time reverse transcription PCR (qRT-PCR) demonstrated the accumulation of mRNAs that encoded glycogen biosynthesis-related enzymes in photoheterotrophic Δ1415 cells. At 96 h, enzyme activity measurement in the photoheterotrophic Δ1415 cells demonstrated that the activities of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase were decreased, but the activities of glucose dehydrogenase were increased. Furthermore, metabolomics analysis demonstrated that the Δ1415 cells showed increased glucose-6-phosphate and 6-phosphogluconate content at 96 h. Therefore, sll1415 has a significant function in the oxidative pentose phosphate (OPP) pathway for catabolism of glucose under photoheterotrophic conditions. Additionally, it is presumed that the slr0400 had a different role in glucose catabolism during growth. These results suggest that the two Synechocystis sp. PCC 6803 NADKs (Sll1415 and Slr0400) have distinct functions in photoheterotrophic cyanobacterial metabolism.
吡啶核苷酸 (NAD(P)(H)) 是电子载体,是各种代谢途径的驱动力。NAD(H) 磷酸化为 NADP(H) 是由酶 NAD 激酶 (NADK) 完成的。集胞藻 PCC 6803 含有两个编码具有 NADK 同源性的蛋白质的基因 (sll1415 和 slr0400)。当 sll1415 和 slr0400 的遗传突变体 (Δ1415 和 Δ0400,分别) 在光异养生长条件下培养时,只有 Δ1415 细胞表现出生长缺陷。在野生型细胞中,细胞转移到光异养条件后,sll1415 转录本积累。此外,NAD(P)(H) 测量表明,在从光自养到光异养的适应过程中,发生了动态的代谢转换。电子显微镜观察和生物化学定量表明,在光异养条件下,Δ1415 细胞在 96 小时时积累糖原。定量实时逆转录 PCR (qRT-PCR) 表明,在光异养 Δ1415 细胞中,编码糖原生物合成相关酶的 mRNAs 积累。在 96 小时时,光异养 Δ1415 细胞的酶活性测量表明,葡萄糖-6-磷酸脱氢酶和 6-磷酸葡萄糖酸脱氢酶的活性降低,但葡萄糖脱氢酶的活性增加。此外,代谢组学分析表明,Δ1415 细胞在 96 小时时显示出葡萄糖-6-磷酸和 6-磷酸葡萄糖酸含量增加。因此,sll1415 在光异养条件下葡萄糖分解的氧化戊糖磷酸 (OPP) 途径中具有重要功能。此外,推测 slr0400 在生长过程中葡萄糖分解中有不同的作用。这些结果表明,两种集胞藻 PCC 6803 NADK (Sll1415 和 Slr0400) 在光异养蓝藻代谢中有不同的功能。