Metabolomic analysis of NAD kinase-deficient mutants of the cyanobacterium Synechocystis sp. PCC 6803

Ishikawa, Yuuma; Miyagi, Atsuko; Haishima, Yuto; Ishikawa, Toshiki; Nagano, Minoru; Yamaguchi, Masatoshi; Hihara, Yukako; Kawai‐Yamada, Maki

doi:10.1016/j.jplph.2016.09.002

Cited by 17 publications

(11 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Metabolic profiling was performed on rodent muscle , heart , liver , brain , intestinal and glioma tissue, arterial cells as well as on rodent serum/plasma , urine and feces . CE‐MS was furthermore used for research on the metabolism of microorganism , plants and mushroom . Additional publications are included in Table .…”

Section: Analytical Applicationsmentioning

confidence: 99%

Recent advances in capillary electrophoresis‐mass spectrometry: Instrumentation, methodology and applications

et al. 2018

View full text Add to dashboard Cite

Capillary electrophoresis (CE) offers fast and high‐resolution separation of charged analytes from small injection volumes. Coupled to mass spectrometry (MS), it represents a powerful analytical technique providing (exact) mass information and enables molecular characterization based on fragmentation. Although hyphenation of CE and MS is not straightforward, much emphasis has been placed on enabling efficient ionization and user‐friendly coupling. Though several interfaces are now commercially available, research on more efficient and robust interfacing with nano‐electrospray ionization (ESI), matrix‐assisted laser desorption/ionization (MALDI) and inductively coupled plasma mass spectrometry (ICP) continues with considerable results. At the same time, CE‐MS has been used in many fields, predominantly for the analysis of proteins, peptides and metabolites. This review belongs to a series of regularly published articles, summarizing 248 articles covering the time between June 2016 and May 2018. Latest developments on hyphenation of CE with MS as well as instrumental developments such as two‐dimensional separation systems with MS detection are mentioned. Furthermore, applications of various CE‐modes including capillary zone electrophoresis (CZE), nonaqueous capillary electrophoresis (NACE), capillary gel electrophoresis (CGE) and capillary isoelectric focusing (CIEF) coupled to MS in biological, pharmaceutical and environmental research are summarized.

show abstract

Section: Analytical Applicationsmentioning

confidence: 99%

Recent advances in capillary electrophoresis‐mass spectrometry: Instrumentation, methodology and applications

et al. 2018

View full text Add to dashboard Cite

show abstract

“…Although the growth curves of the WT Synechocystis strain and the NADK‐deficient mutants (Δ1415 and Δ0400) did not differ under photoautotrophic conditions, the metabolomics profile of Δ0400 differed from those of WT and Δ1415 (Ishikawa et al ., ). Therefore, it was hypothesized that sll1415 and slr0400 may possess metabolically distinct roles under photoautotrophic conditions.…”

Section: Discussionmentioning

confidence: 97%

“…PCC 6803 was used as the wild‐type (WT) strain. NADK‐deficient mutants were obtained by inserting an antibiotic‐resistance‐conferring cassette in either of the two NADK genes; detailed information on these constructions is provided in our previous report (Ishikawa et al ., ). For photoautotrophy, strains were grown in BG11 culture medium (20 ml) on a shaker (100 rpm) at 30°C under continuous light at 30 μE m −2 sec −1 .…”

Section: Methodsmentioning

confidence: 97%

“…PCC 6803 single mutants lacking one of these genes (Δ1415 or Δ0400) and performed metabolomics analyses under photoautotrophic conditions. Despite the similar growth properties of the wild‐type (WT) strain and the single‐mutant strains under autotrophic conditions, only Δ0400 showed a metabolite profile that differed from that of WT (Ishikawa et al ., ). However, the details of the metabolic alterations associated with the mutations of these genes ( sll1415 and slr0400) remain unclear and the biological functions of the two NADK isoforms in Synechocystis are unknown.…”

Section: Introductionmentioning

confidence: 97%

See 1 more Smart Citation

One of the NAD kinases, sll1415, is required for the glucose metabolism of Synechocystis sp. PCC 6803

et al. 2019

Self Cite

View full text Add to dashboard Cite

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 (D1415 and D0400, respectively) were cultured under photoheterotrophic growth conditions only the D1415 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 D1415 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 D1415 cells. At 96 h, enzyme activity measurement in the photoheterotrophic D1415 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 D1415 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.

show abstract

“…On the other hands, there was no apparent difference between the pntAB -deficient mutant and WT under the mixotrophic condition (20 μE/m 2 /s). Interestingly, sll1415 -deficient mutant, one of the NADK -deficient mutants in Synechocystis , and pntAB -deficient mutant showed the similar phenotype under the day-night rhythm in the presence of glucose and the mixotrophic condition (20 μE/m 2 /s) (Ishikawa et al, 2016, 2019; Kamarainen et al, 2017). Therefore, pntAB may maintain the NADPH pool with sll1415 by unknown coordination system.…”

Section: Nad Kinases In Cyanobacteriamentioning

confidence: 93%

Physiological Significance of NAD Kinases in Cyanobacteria

Ishikawa

Kawai‐Yamada

2019

Front. Plant Sci.

Self Cite

View full text Add to dashboard Cite

Unicellular cyanobacteria are thought to be the evolutionary ancestors of plant chloroplasts and are widely used both for chemical production and as model organisms in studies of photosynthesis. Although most research focused on increasing reducing power (that is, NADPH) as target of metabolic engineering, the physiological roles of NAD(P)(H) in cyanobacteria poorly understood. In cyanobacteria such as the model species Synechocystis sp. PCC 6803, most metabolic pathways share a single compartment. This complex metabolism raises the question of how cyanobacteria control the amounts of the redox pairs NADH/NAD + and NADPH/NADP + in the cyanobacterial metabolic pathways. For example, photosynthetic and respiratory electron transport chains share several redox components in the thylakoid lumen, including plastoquinone, cytochrome b 6 f (cyt b 6 f ), and the redox carriers plastocyanin and cytochrome c6 . In the case of photosynthesis, NADP + acts as an important electron mediator on the acceptor-side of photosystem I (PSI) in the linear electron chain as well as in the plant chloroplast. Meanwhile, in respiration, most electrons derived from NADPH and NADH are transferred by NAD(P)H dehydrogenases. Therefore, it is expected that Synechocystis employs unique NAD(P)(H) -pool control mechanisms to regulate the mixed metabolic systems involved in photosynthesis and respiration. This review article summarizes the current state of knowledge of NAD(P)(H) metabolism in Synechocystis . In particular, we focus on the physiological function in Synechocystis of NAD kinase, the enzyme that phosphorylates NAD + to NADP + .

show abstract

Metabolomic analysis of NAD kinase-deficient mutants of the cyanobacterium Synechocystis sp. PCC 6803

Cited by 17 publications

References 33 publications

Recent advances in capillary electrophoresis‐mass spectrometry: Instrumentation, methodology and applications

Recent advances in capillary electrophoresis‐mass spectrometry: Instrumentation, methodology and applications

One of the NAD kinases, sll1415, is required for the glucose metabolism of Synechocystis sp. PCC 6803

Physiological Significance of NAD Kinases in Cyanobacteria

Contact Info

Product

Resources

About