Genome-Based Metabolic Mapping and 13C Flux Analysis Reveal Systematic Properties of an Oleaginous Microalga Chlorella protothecoides

Wu, Chao; Xiong, Wei; Dai, Junbiao; Wu, Qingyu

doi:10.1104/pp.114.250688

Title: Genome-Based Metabolic Mapping and ¹³C Flux Analysis Reveal Systematic Properties of an Oleaginous Microalga Chlorella protothecoides

Journal Article · Mon Dec 15 00:00:00 EST 2014 · Plant Physiology (Bethesda)

DOI:https://doi.org/10.1104/pp.114.250688· OSTI ID:1220638

Wu, Chao ^[1]; Xiong, Wei ^[1]; Dai, Junbiao ^[1]; Wu, Qingyu ^[1]

Tsinghua Univ., Beijing (China). Ministry of Education Key Lab. of Bioinformatics; National Renewable Energy Lab. (NREL), Golden, CO (United States). Biosciences Center

We report that integrated and genome-based flux balance analysis, metabolomics, and ¹³C-label profiling of phototrophic and heterotrophic metabolism in Chlorella protothecoides, an oleaginous green alga for biofuel. The green alga Chlorella protothecoides, capable of autotrophic and heterotrophic growth with rapid lipid synthesis, is a promising candidate for biofuel production. Based on the newly available genome knowledge of the alga, we reconstructed the compartmentalized metabolic network consisting of 272 metabolic reactions, 270 enzymes, and 461 encoding genes and simulated the growth in different cultivation conditions with flux balance analysis. Phenotype-phase plane analysis shows conditions achieving theoretical maximum of the biomass and corresponding fatty acid-producing rate for phototrophic cells (the ratio of photon uptake rate to CO₂ uptake rate equals 8.4) and heterotrophic ones (the glucose uptake rate to O₂ consumption rate reaches 2.4), respectively. Isotope-assisted liquid chromatography-mass spectrometry/mass spectrometry reveals higher metabolite concentrations in the glycolytic pathway and the tricarboxylic acid cycle in heterotrophic cells compared with autotrophic cells. We also observed enhanced levels of ATP, nicotinamide adenine dinucleotide (phosphate), reduced, acetyl-Coenzyme A, and malonyl-Coenzyme A in heterotrophic cells consistently, consistent with a strong activity of lipid synthesis. To profile the flux map in experimental conditions, we applied nonstationary ¹³C metabolic flux analysis as a complementing strategy to flux balance analysis. We found that the result reveals negligible photorespiratory fluxes and a metabolically low active tricarboxylic acid cycle in phototrophic C. protothecoides. In comparison, high throughput of amphibolic reactions and the tricarboxylic acid cycle with no glyoxylate shunt activities were measured for heterotrophic cells. Lastly, taken together, the metabolic network modeling assisted by experimental metabolomics and ¹³C labeling better our understanding on global metabolism of oleaginous alga, paving the way to the systematic engineering of the microalga for biofuel production.

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Research Organization:: National Renewable Energy Laboratory (NREL), Golden, CO (United States)

Sponsoring Organization:: NREL Director's Postdoc Fellowship; National Natural Science Foundation of China (NSFC)

Grant/Contract Number:: AC36-08GO28308; 31370282; 41030210; 2012Z08128

OSTI ID:: 1220638

Report Number(s):: NREL/JA-2700-63117

Journal Information:: Plant Physiology (Bethesda), Vol. 167, Issue 2; ISSN 0032-0889

Publisher:: American Society of Plant BiologistsCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 71 works

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References (56)

Ion Suppression in Mass Spectrometry Annesley, T. M. Clinical Chemistry, Vol. 49, Issue 7 https://doi.org/10.1373/49.7.1041	journal	July 2003
Elementary metabolite units (EMU): A novel framework for modeling isotopic distributions Antoniewicz, Maciek R.; Kelleher, Joanne K.; Stephanopoulos, Gregory Metabolic Engineering, Vol. 9, Issue 1 https://doi.org/10.1016/j.ymben.2006.09.001	journal	January 2007
Intermediates in the photosynthetic cycle Bassham, J. A.; Barker, S. A.; Calvin, M. Biochimica et Biophysica Acta, Vol. 21, Issue 2 https://doi.org/10.1016/0006-3002(56)90022-1	journal	August 1956
The path of Carbon in Photosynthesis Bassham, James A.; Benson, Andrew A.; Calvin, Melvin. Journal of Biological Chemistry, Vol. 185, Issue 2 https://doi.org/10.1016/S0021-9258(18)56368-7	journal	August 1950
Absolute quantitation of intracellular metabolite concentrations by an isotope ratio-based approach Bennett, Bryson D.; Yuan, Jie; Kimball, Elizabeth H. Nature Protocols, Vol. 3, Issue 8 https://doi.org/10.1038/nprot.2008.107	journal	July 2008
A simple and reliable method for the determination of cellular RNA content Benthin, S.; Nielsen, J.; Villadsen, J. Biotechnology Techniques, Vol. 5, Issue 1 https://doi.org/10.1007/BF00152753	journal	January 1991
Flux balance analysis of primary metabolism in Chlamydomonas reinhardtii Boyle, Nanette R.; Morgan, John A. BMC Systems Biology, Vol. 3, Issue 1 https://doi.org/10.1186/1752-0509-3-4	journal	January 2009
Metabolic network reconstruction of Chlamydomonas offers insight into light‐driven algal metabolism Chang, Roger L.; Ghamsari, Lila; Manichaikul, Ani Molecular Systems Biology, Vol. 7, Issue 1 https://doi.org/10.1038/msb.2011.52	journal	January 2011
Synergy between 13C-metabolic flux analysis and flux balance analysis for understanding metabolic adaption to anaerobiosis in E. coli Chen, Xuewen; Alonso, Ana P.; Allen, Doug K. Metabolic Engineering, Vol. 13, Issue 1 https://doi.org/10.1016/j.ymben.2010.11.004	journal	January 2011
Bioassay-guided purification and identification of PPARα/γ agonists fromChlorella sorokiniana Chou, Yu-Cheng; Prakash, Ekambaranellore; Huang, Chien-Fu Phytotherapy Research, Vol. 22, Issue 5 https://doi.org/10.1002/ptr.2280	journal	January 2008
Enzymic evidence for peroxisomes in a mutant of Chlorella vulgaris Codd, Geoffrey A.; Schmid, Georg H.; Kowallik, Wolfgang Archiv f�r Mikrobiologie, Vol. 81, Issue 3 https://doi.org/10.1007/BF00412245	journal	January 1972
Identification of a metabolic network structure representative ofarthrospira (spirulina) platensis metabolism Cogne, Guillaume; Gros, J. -B.; Dussap, C. -G. Biotechnology and Bioengineering, Vol. 84, Issue 6 https://doi.org/10.1002/bit.10808	journal	October 2003
Regulation of citrate synthase activity ofSaccharomyces cerevisiae Coleman, J. S.; Bhattacharjee, J. K. Antonie van Leeuwenhoek, Vol. 41, Issue 1 https://doi.org/10.1007/BF02565060	journal	December 1975
Metabolic modelling of microbes: the flux-balance approach Edwards, Jeremy S.; Covert, Markus; Palsson, Bernhard Environmental Microbiology, Vol. 4, Issue 3 https://doi.org/10.1046/j.1462-2920.2002.00282.x	journal	March 2002
In silico predictions of Escherichia coli metabolic capabilities are consistent with experimental data Edwards, Jeremy S.; Ibarra, Rafael U.; Palsson, Bernhard O. Nature Biotechnology, Vol. 19, Issue 2 https://doi.org/10.1038/84379	journal	February 2001
Cluster analysis and display of genome-wide expression patterns Eisen, M. B.; Spellman, P. T.; Brown, P. O. Proceedings of the National Academy of Sciences, Vol. 95, Issue 25 https://doi.org/10.1073/pnas.95.25.14863	journal	December 1998
Genome-scale modeling of Synechocystis sp. PCC 6803 and prediction of pathway insertion Fu, Pengcheng Journal of Chemical Technology & Biotechnology, Vol. 84, Issue 4 https://doi.org/10.1002/jctb.2065	journal	April 2009
Oil accumulation mechanisms of the oleaginous microalga Chlorella protothecoides revealed through its genome, transcriptomes, and proteomes Gao, Chunfang; Wang, Yun; Shen, Yue BMC Genomics, Vol. 15, Issue 1 https://doi.org/10.1186/1471-2164-15-582	journal	January 2014
Diphenylamine-colorimetric method for DNA assay: A shortened procedure by incubating samples at 50°C Gendimenico, Gerard J.; Bouquin, Pamela L.; Tramposch, Kenneth M. Analytical Biochemistry, Vol. 173, Issue 1 https://doi.org/10.1016/0003-2697(88)90156-X	journal	August 1988
Microalgae as a raw material for biofuels production Gouveia, Luisa; Oliveira, Ana Cristina Journal of Industrial Microbiology & Biotechnology, Vol. 36, Issue 2, p. 269-274 https://doi.org/10.1007/s10295-008-0495-6	journal	November 2008
The Respiratory Chain of Chlorella protothecoides: I. Inhibitor Responses and Cytochrome Components of Whole Cells Grant, Neil G.; Hommersand, Max H. Plant Physiology, Vol. 54, Issue 1 https://doi.org/10.1104/pp.54.1.50	journal	July 1974
Metabolic network modeling of redox balancing and biohydrogen production in purple nonsulfur bacteria Hädicke, Oliver; Grammel, Hartmut; Klamt, Steffen BMC Systems Biology, Vol. 5, Issue 1 https://doi.org/10.1186/1752-0509-5-150	journal	January 2011
AMP-activated protein kinase: an ultrasensitive system for monitoring cellular energy charge Hardie, D. Grahame; Salt, Ian P.; Hawley, Simon A. Biochemical Journal, Vol. 338, Issue 3 https://doi.org/10.1042/bj3380717	journal	March 1999
Modulation of the Major Paths of Carbon in Photorespiratory Mutants of Synechocystis Huege, Jan; Goetze, Jan; Schwarz, Doreen PLoS ONE, Vol. 6, Issue 1 https://doi.org/10.1371/journal.pone.0016278	journal	January 2011
KEGG: Kyoto Encyclopedia of Genes and Genomes Kanehisa, Minoru; Goto, Susumu Nucleic Acids Research, Vol. 28, Issue 1, p. 27-30 https://doi.org/10.1093/nar/28.1.27	journal	January 2000
Structural and functional analysis of cellular networks with CellNetAnalyzer Klamt, Steffen; Saez-Rodriguez, Julio; Gilles, Ernst D. BMC Systems Biology, Vol. 1, Issue 1 https://doi.org/10.1186/1752-0509-1-2	journal	January 2007
iAK692: A genome-scale metabolic model of Spirulina platensis C1 Klanchui, Amornpan; Khannapho, Chiraphan; Phodee, Atchara BMC Systems Biology, Vol. 6, Issue 1 https://doi.org/10.1186/1752-0509-6-71	journal	January 2012
The Metabolic Network of Synechocystis sp. PCC 6803: Systemic Properties of Autotrophic Growth Knoop, Henning; Zilliges, Yvonne; Lockau, Wolfgang Plant Physiology, Vol. 154, Issue 1 https://doi.org/10.1104/pp.110.157198	journal	July 2010
Flux balance analysis in the era of metabolomics Lee, J. M. Briefings in Bioinformatics, Vol. 7, Issue 2 https://doi.org/10.1093/bib/bbl007	journal	March 2006
Microalgal mass culture systems and methods: Their limitation and potential Lee, Yuan-Kun Journal of Applied Phycology, Vol. 13, Issue 4, p. 307-315 https://doi.org/10.1023/A:1017560006941	journal	August 2001
Metabolic network analysis integrated with transcript verification for sequenced genomes Manichaikul, Ani; Ghamsari, Lila; Hom, Erik F. Y. Nature Methods, Vol. 6, Issue 8 https://doi.org/10.1038/nmeth.1348	journal	July 2009
Matrix Effect in Quantitative LC/MS/MS Analyses of Biological Fluids: A Method for Determination of Finasteride in Human Plasma at Picogram Per Milliliter Concentrations Matuszewski, B. K.; Constanzer, M. L.; Chavez-Eng, C. M. Analytical Chemistry, Vol. 70, Issue 5 https://doi.org/10.1021/ac971078+	journal	February 1998
Biodiesel production from heterotrophic microalgal oil Miao, Xiaoling; Wu, Qingyu Bioresource Technology, Vol. 97, Issue 6 https://doi.org/10.1016/j.biortech.2005.04.008	journal	April 2006
Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar Miller, G. L. Analytical Chemistry, Vol. 31, Issue 3 https://doi.org/10.1021/ac60147a030	journal	March 1959
Reconstruction and analysis of genome-scale metabolic model of a photosynthetic bacterium Montagud, Arnau; Navarro, Emilio; Fernández de Córdoba, Pedro BMC Systems Biology, Vol. 4, Issue 1 https://doi.org/10.1186/1752-0509-4-156	journal	November 2010
Systems-level metabolic flux profiling identifies fatty acid synthesis as a target for antiviral therapy Munger, Joshua; Bennett, Bryson D.; Parikh, Anuraag Nature Biotechnology, Vol. 26, Issue 10 https://doi.org/10.1038/nbt.1500	journal	September 2008
Detailing the optimality of photosynthesis in cyanobacteria through systems biology analysis Nogales, J.; Gudmundsson, S.; Knight, E. M. Proceedings of the National Academy of Sciences, Vol. 109, Issue 7 https://doi.org/10.1073/pnas.1117907109	journal	January 2012
Applications of genome‐scale metabolic reconstructions Oberhardt, Matthew A.; Palsson, Bernhard Ø; Papin, Jason A. Molecular Systems Biology, Vol. 5, Issue 1 https://doi.org/10.1038/msb.2009.77	journal	January 2009
The Incomplete Tricarboxylic Acid Cycle in the Blue-green Alga Anabaena Variabilis Pearce, J.; Leach, C. K.; Carr, N. G. Journal of General Microbiology, Vol. 55, Issue 3 https://doi.org/10.1099/00221287-55-3-371	journal	March 1969
Valuable products from biotechnology of microalgae Pulz, Otto; Gross, Wolfgang Applied Microbiology and Biotechnology, Vol. 65, Issue 6 https://doi.org/10.1007/s00253-004-1647-x	journal	August 2004
Inhibition of photosynthesis by intracellular carbonic anhydrase in microalgae under excess concentrations of CO2 Satoh, Akira; Kurano, Norihide; Miyachi, Shigetoh Photosynthesis Research, Vol. 68, Issue 3, p. 215-224 https://doi.org/10.1023/A:1012980223847	journal	January 2001
Recent Applications of Metabolomics Toward Cyanobacteria Schwarz, Doreen; Orf, Isabel; Kopka, Joachim Metabolites, Vol. 3, Issue 1 https://doi.org/10.3390/metabo3010072	journal	February 2013
Quantitative analysis of metabolites in complex biological samples using ion-pair reversed-phase liquid chromatography–isotope dilution tandem mass spectrometry Seifar, Reza M.; Zhao, Zheng; van Dam, Jan Journal of Chromatography A, Vol. 1187, Issue 1-2 https://doi.org/10.1016/j.chroma.2008.02.008	journal	April 2008
Flux Balance Analysis of Photoautotrophic Metabolism Shastri, A. A.; Morgan, J. A. Biotechnology Progress, Vol. 21, Issue 6 https://doi.org/10.1021/bp050246d	journal	December 2005
Maximum Growth Rates of Chlorella in Steady-State and in Synchronized Cultures Sorokin, C.; Krauss, R. W. Proceedings of the National Academy of Sciences, Vol. 45, Issue 12 https://doi.org/10.1073/pnas.45.12.1740	journal	December 1959
Unusual cyanobacterial TCA cycles: not broken just different Steinhauser, Dirk; Fernie, Alisdair R.; Araújo, Wagner L. Trends in Plant Science, Vol. 17, Issue 9 https://doi.org/10.1016/j.tplants.2012.05.005	journal	September 2012
PredAlgo: A New Subcellular Localization Prediction Tool Dedicated to Green Algae Tardif, Marianne; Atteia, Ariane; Specht, Michael Molecular Biology and Evolution, Vol. 29, Issue 12 https://doi.org/10.1093/molbev/mss178	journal	July 2012
Fine control of citrate synthase activity in blue-green algae Taylor, Barrie F. Archiv f�r Mikrobiologie, Vol. 92, Issue 3 https://doi.org/10.1007/BF00411205	journal	January 1973
Flux through Citrate Synthase Limits the Growth of Ethanologenic Escherichia coli KO11 during Xylose Fermentation Underwood, S. A.; Buszko, M. L.; Shanmugam, K. T. Applied and Environmental Microbiology, Vol. 68, Issue 3 https://doi.org/10.1128/AEM.68.3.1071-1081.2002	journal	March 2002
Double CO2 fixation in photosynthesis–fermentation model enhances algal lipid synthesis for biodiesel production Xiong, Wei; Gao, Chunfang; Yan, Dong Bioresource Technology, Vol. 101, Issue 7 https://doi.org/10.1016/j.biortech.2009.11.041	journal	April 2010
High-density fermentation of microalga Chlorella protothecoides in bioreactor for microbio-diesel production Xiong, Wei; Li, Xiufeng; Xiang, Jinyi Applied Microbiology and Biotechnology, Vol. 78, Issue 1 https://doi.org/10.1007/s00253-007-1285-1	journal	February 2008
¹³ C-Tracer and Gas Chromatography-Mass Spectrometry Analyses Reveal Metabolic Flux Distribution in the Oleaginous Microalga Chlorella protothecoides Xiong, Wei; Liu, Lixia; Wu, Chao Plant Physiology, Vol. 154, Issue 2 https://doi.org/10.1104/pp.110.158956	journal	August 2010
High quality biodiesel production from a microalga Chlorella protothecoides by heterotrophic growth in fermenters Xu, Han; Miao, Xiaoling; Wu, Qingyu Journal of Biotechnology, Vol. 126, Issue 4 https://doi.org/10.1016/j.jbiotec.2006.05.002	journal	December 2006
INCA: a computational platform for isotopically non-stationary metabolic flux analysis Young, J. D. Bioinformatics, Vol. 30, Issue 9 https://doi.org/10.1093/bioinformatics/btu015	journal	January 2014
Mapping photoautotrophic metabolism with isotopically nonstationary 13C flux analysis Young, Jamey D.; Shastri, Avantika A.; Stephanopoulos, Gregory Metabolic Engineering, Vol. 13, Issue 6 https://doi.org/10.1016/j.ymben.2011.08.002	journal	November 2011
An elementary metabolite unit (EMU) based method of isotopically nonstationary flux analysis Young, Jamey D.; Walther, Jason L.; Antoniewicz, Maciek R. Biotechnology and Bioengineering, Vol. 99, Issue 3 https://doi.org/10.1002/bit.21632	journal	February 2008

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Title: Genome-Based Metabolic Mapping and 13C Flux Analysis Reveal Systematic Properties of an Oleaginous Microalga Chlorella protothecoides

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Title: Genome-Based Metabolic Mapping and ¹³C Flux Analysis Reveal Systematic Properties of an Oleaginous Microalga Chlorella protothecoides