Cited by 241Microbial Metabolic Engineering and BioproductionPhotosynthetic Processes and MechanismsAlgal biology and biofuel production

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Metabolic Flux Analysis in Synechocystis Using Isotope Distribution from 13C-Labeled Glucose

Yang, Chen, Hua, Qiang, Shimizu, Kazuyuki·2002·Metabolic Engineering

DOI 10.1006/mben.2002.0226

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Abstract

Using the carbon isotope labeling technique, the response of cyanobacterial central carbon metabolism to the change in environmental conditions was investigated. Synechocystis was grown in the heterotrophic and mixotrophic cultures fed with 13C-labeled glucose. The labeling patterns of the amino acids in biomass hydrolysates for both cultures were detected by the two-dimensional 1H-13C correlation nuclear magnetic resonance (2D 1H-13C COSY NMR) spectroscopy and gas chromatography-mass spectrometry (GC-MS) technique. The in vivo intracellular flux distributions were then quantitated from the labeling measurements and metabolite balances using a parameters fitting approach. From the estimated flux distributions, it was found that the pentose phosphate pathway was the major pathway of glucose catabolism in the heterotrophic culture, while in the mixotrophic culture, the flux of CO2 fixation through the Calvin cycle was about two-fold of the glucose input flux. The relative flux through the phosphoenolpyruvate carboxylase was very high in both cultures, and this reaction represented about 25% of the assimilated CO2 in the mixotrophic culture. More importantly, we found a substantial outflow from the tricarboxylic acid cycle to glycolysis pathway carried by the malic enzyme, demonstrating the operation of a C4 pathway in cyanobacterial cells through the PEP carboxylase and malic enzyme. The estimated flux distributions also revealed that the NADPH synthesis was in excess relative to its requirement, and the excess NADPH might be reoxidized in cyanobacterial respiration to provide the energy for cellular requirement. Moreover, the analyzed result also suggested that the activity of the respiratory electron transport chain in cyanobacterial cells was not inhibited by light.

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Authors: Yang, C., Hua, Q., & Shimizu, K.
Affiliations: Institute for Advanced Biosciences, Keio University (JP)
Kyushu Institute of Technology (JP)
Corresponding author: Shimizu, Kazuyuki
ORCID: Hua, Qiang
Shimizu, Kazuyuki
Venue: Metabolic Engineering
Publisher: Elsevier BV
Published: 2002-07
Volume: 4
Issue: 3
Pages: 202-216
ISSN: 1096-7176 (ISSN-L)
DOI: 10.1006/mben.2002.0226
OpenAlex: W1490314836
Cited by: 241
References: 41
Citations by year: 2026: 3 · 2025: 9 · 2024: 13 · 2023: 14 · 2022: 11 · 2021: 6 · 2020: 12 · 2019: 8 · 2018: 18 · 2017: 17 · 2016: 15 · 2015: 11 · 2014: 11 · 2013: 18 · 2012: 12
FWCI: 1.51
Type: journal article
Language: en
Open access: closed
Licence: www.elsevier.com/tdm/userlicense/1.0/ (tdm)
www.elsevier.com/legal/tdmrep-license (tdm)
Locations: Metabolic Engineering (publishedVersion)
PubMed (publishedVersion)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Photosynthetic Processes and Mechanisms
Physical Sciences › Energy › Renewable Energy, Sustainability and the Environment › Algal biology and biofuel production
Keywords: Synechocystis, metabolic flux distribution, 13C-labeled glucose, pentose phosphate pathway, Calvin cycle, phosphoenolpyruvate carboxylase, malic enzyme, C4 photosynthesis, TCA cycle, NADPH production, heterotrophic culture, mixotrophic cultivation, CO2 fixation, respiratory electron transport chain
MeSH terms: Cyanobacteria, Amino Acids, Carbon, Carbon Radioisotopes, Cells, Cultured, Computer Simulation, Culture Media, Glucose, Hydrolysis, Isotope Labeling, Gas Chromatography-Mass Spectrometry, Models, Biological, Models, Chemical, Multienzyme Complexes, Species Specificity, Stereoisomerism, Nuclear Magnetic Resonance, Biomolecular

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Metabolic Flux Analysis in Synechocystis Using Isotope Distribution from 13C-Labeled Glucose

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