Abstract
Cyanobacteria accumulate polyglucan as main carbohydrate storage. Here, the cellular polyglucan content was determined in 27 cyanobacterial strains from 25 genera. The polyglucan contents were significantly enhanced in 20 and 23 strains under nitrogen (–N) and phosphate (–P) deprivation, respectively. High polyglucan accumulation was not associated with particular evolutionary groups but was strain specific. The highest polyglucan accumulations of 46.2% and 52.5% (w/w dry weight; DW) were obtained under –N in Synechocystis sp. PCC 6803 (hereafter Synechocystis) and Chroococcus limneticus, respectively. In Synechocystis, 80–97% (w/w) of the polyglucan was glycogen. Transcriptome and metabolome analyses during glycogen accumulation under –N were determined in Synechocystis. The genes responsible for the supply of the substrates for glycogen synthesis: glycerate-1,3-phosphate and fructose-1,6-phosphate, were significantly up-regulated. The genes encoding the enzymes converting succinate to malate in TCA cycle, were significantly down-regulated. The genes encoding the regulator proteins which inhibits metabolism at lower part of glycolysis pathway, were also significantly up-regulated. The transcript levels of PII protein and the level of 2-oxoglutarate, which form a complex that inhibits lower part of glycolysis pathway, were significantly increased. Thus, the increased Synechocystis glycogen accumulation under –N was likely to be mediated by the increased supply of glycogen synthesis substrates and metabolic inhibitions at lower part of glycolysis pathway and TCA cycle.
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Data availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
References
Aikawa S, Ho SH, Nakanishi A, Chang JS, Hasunuma T, Kondo A (2015) Improving polyglucan production in cyanobacteria and microalgae via cultivation design and metabolic engineering. Biotechnol J 10:886–898. doi:https://doi.org/10.1002/biot.201400344
Aikawa S, Izumi Y, Matsuda F, Hasunuma T, Chang J-S, Kondo A (2012) Synergistic enhancement of glycogen production in Arthrospira platensis by optimization of light intensity and nitrate supply. Bioresour Technol 108:211–215. doi:https://doi.org/10.1016/j.biortech.2012.01.004
Aikawa S, Nishida A, Ho S-H, Chang J-S, Hasunuma T, Kondo A (2014) Glycogen production for biofuels by the euryhaline cyanobacteria Synechococcus sp. strain PCC 7002 from an oceanic environment. Biotechnol Biofuels 7:88. doi:https://doi.org/10.1186/1754-6834-7-88
Allwood JW, Weber RJM, Zhou J, He S, Viant MR, Dunn WB (2013) CASMI— the small molecule identification process from a Birmingham perspective. Metabolites 3:397–411. doi:https://doi.org/10.3390/metabo3020397
Arias DM, García J, Uggetti E (2020) Production of polymers by cyanobacteria grown in wastewater: current status, challenges and future perspectives. New Biotechnol 55:46–57. doi:https://doi.org/10.1016/j.nbt.2019.09.001
Badary A, Takamatsu S, Nakajima M, Ferri S, Lindblad P, Sode K (2018) Glycogen production in marine cyanobacterial strain Synechococcus sp. NKBG 15041c. Mar Biotechnol 20:109–117. doi:https://doi.org/10.1007/s10126-017-9792-2
Ball SG, Morell MK (2003) From bacterial glycogen to starch: understanding the biogenesis of the plant starch granule. Annu Rev Plant Biol 54:207–233. doi:https://doi.org/10.1146/annurev.arplant.54.031902.134927
Collier JL, Grossman AR (1994) A small polypeptide triggers complete degradation of light-harvesting phycobiliproteins in nutrient-deprived cyanobacteria. EMBO J 13:1039–1047. doi:https://doi.org/10.1002/j.1460-2075.1994.tb06352.x
De Farias Silva CE, Sforza E, Bertucco A (2017) Effects of pH and carbon source on Synechococcus PCC 7002 cultivation: biomass and carbohydrate production with different strategies for pH control. Appl Biochem Biotechnol 181:682–698. doi:https://doi.org/10.1007/s12010-016-2241-2
De Philippis R, Sili C, Vincenzini M (1992) Glycogen and poly-β-hydroxybutyrate synthesis in Spirulina maxima. J Gen Microbiol 138:1623–1628. doi:https://doi.org/10.1099/00221287-138-8-1623
De Winder B, Stal LJ, Mur LR (1990) Crinalium epipsammum sp. nov.: a filamentous cyanobacterium with trichomes composed of elliptical cells and containing poly-β-(1, 4) glucar (cellulose). J Gen Microbiol 136:1645–1653. doi:https://doi.org/10.1099/00221287-136-8-1645
Ernst A, Kirschenlohr H, Diez J, Böger P (1984) Glycogen content and nitrogenase activity in Anabaena variabilis. Arch Microbiol 140:120–125. doi:https://doi.org/10.1007/BF00454913
Forchhammer K, Selim KA, Huergo LF (2022) New views on PII signaling: from nitrogen sensing to global metabolic control. Trends Microbiol. doi:https://doi.org/10.1016/j.tim.2021.12.014
Giner-Lamia J, Robles-Rengel R, Hernández-Prieto MA, Muro-Pastor MI, Florencio FJ, Futschik ME (2017) Identification of the direct regulon of NtcA during early acclimation to nitrogen starvation in the cyanobacterium Synechocystis sp. PCC 6803. Nucleic Acids Res 45:11800–11820. doi:https://doi.org/10.1093/nar/gkx860
González López CV, García MdCC, Fernández FGA, Bustos CS, Chisti Y, Sevilla JMF (2010) Protein measurements of microalgal and cyanobacterial biomass. Bioresour Technol 101:7587–7591. doi:https://doi.org/10.1016/j.biortech.2010.04.077
Gründel M, Scheunemann R, Lockau W, Zilliges Y (2012) Impaired glycogen synthesis causes metabolic overflow reactions and affects stress responses in the cyanobacterium Synechocystis sp. PCC 6803. Microbiology 158:3032–3043. doi:https://doi.org/10.1099/mic.0.062950-0
Guijas C, Montenegro-Burke JR, Domingo-Almenara X, Palermo A, Warth B, Hermann G, Koellensperger G, Huan T, Uritboonthai W, Aisporna AE, Wolan DW, Spilker ME, Benton HP, Siuzdak G (2018) METLIN: a technology platform for identifying knowns and unknowns. Anal Chem 90:3156–3164. doi:https://doi.org/10.1021/acs.analchem.7b04424
Hasunuma T, Kikuyama F, Matsuda M, Aikawa S, Izumi Y, Kondo A (2013) Dynamic metabolic profiling of cyanobacterial glycogen biosynthesis under conditions of nitrate depletion. J Exp Bot 64:2943–2954. doi:https://doi.org/10.1093/jxb/ert134
Huang S, Chen L, Te R, Qiao J, Wang J, Zhang W (2013) Complementary iTRAQ proteomics and RNA-seq transcriptomics reveal multiple levels of regulation in response to nitrogen starvation in Synechocystis sp. PCC 6803. Mol BioSyst 9:2565–2574. doi:https://doi.org/10.1039/c3mb70188c
Ito S, Osanai T (2020) Unconventional biochemical regulation of the oxidative pentose phosphate pathway in the model cyanobacterium Synechocystis sp. PCC 6803. Biochem J 477:1309–1321. doi:https://doi.org/10.1042/bcj20200038
Kanehisa M, Goto S (2000) KEGG: kyoto encyclopedia of genes and genomes. Nucleic Acids Res 28:27–30. doi:https://doi.org/10.1093/nar/28.1.27
Karradt A, Sobanski J, Mattow J, Lockau W, Baier K (2008) NblA, a key protein of phycobilisome degradation, interacts with ClpC, a HSP100 chaperone partner of a cyanobacterial clp protease. J Biol Chem 283:32394–32403. doi:https://doi.org/10.1074/jbc.M805823200
Kawano Y, Saotome T, Ochiai Y, Katayama M, Narikawa R, Ikeuchi M (2011) Cellulose accumulation and a cellulose synthase gene are responsible for cell aggregation in the cyanobacterium Thermosynechococcus vulcanus RKN. Plant Cell Physiol 52:957–966. doi:https://doi.org/10.1093/pcp/pcr047
Kiyota H, Hirai MY, Ikeuchi M (2014) NblA1/A2-dependent homeostasis of amino acid pools during nitrogen starvation in Synechocystis sp. PCC 6803. Metabolites 4:517–531. doi:https://doi.org/10.3390/metabo4030517
Klotz A, Georg J, Bučinská L, Watanabe S, Reimann V, Januszewski W, Sobotka R, Jendrossek D, Hess Wolfgang R, Forchhammer K (2016) Awakening of a dormant cyanobacterium from nitrogen chlorosis reveals a genetically determined program. Curr Biol 26:2862–2872. doi:https://doi.org/10.1016/j.cub.2016.08.054
Knoop H, Gründel M, Zilliges Y, Lehmann R, Hoffmann S, Lockau W, Steuer R (2013) Flux balance analysis of cyanobacterial metabolism: the metabolic network of Synechocystis sp. PCC 6803. PLoS Comput Biol 9:e1003081. doi:https://doi.org/10.1371/journal.pcbi.1003081
Knoop H, Zilliges Y, Lockau W, Steuer R (2010) The metabolic network of Synechocystis sp. PCC 6803: systemic properties of autotrophic growth. Plant Physiol 154:410–422. doi:https://doi.org/10.1104/pp.110.157198
Komárek J, Kaštovský J, Mareš J, Johansen JR (2014) Taxonomic classification of cyanoprokaryotes (cyanobacterial genera) 2014, using a polyphasic approach. Preslia 86:295–335
Kozlov AM, Zhang J, Yilmaz P, Glöckner FO, Stamatakis A (2016) Phylogeny-aware identification and correction of taxonomically mislabeled sequences. Nucleic Acids Res 44:5022–5033. doi:https://doi.org/10.1093/nar/gkw396
Kushwaha D, Upadhyay SN, Mishra PK (2018) Growth of cyanobacteria: optimization for increased carbohydrate content. Appl Biochem Biotechnol 184:1247–1262. doi:https://doi.org/10.1007/s12010-017-2620-3
Liu D, Yang C (2014) The nitrogen-regulated response regulator NrrA controls cyanophycin synthesis and glycogen catabolism in the cyanobacterium Synechocystis sp. PCC 6803. J Biol Chem 289:2055–2071. doi:https://doi.org/10.1074/jbc.M113.515270
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2–∆∆CT method. Methods 25:402–408. doi:https://doi.org/10.1006/meth.2001.1262
Llácer JL, Espinosa J, Castells MA, Contreras A, Forchhammer K, Rubio V (2010) Structural basis for the regulation of NtcA-dependent transcription by proteins PipX and PII. Proc Natl Acad Sci U S A 107:15397–15402. doi:https://doi.org/10.1073/pnas.1007015107
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275. doi:https://doi.org/10.1016/S0021-9258(19)52451-6
Markou G (2012) Alteration of the biomass composition of Arthrospira (Spirulina) platensis under various amounts of limited phosphorus. Bioresour Technol 116:533–535. doi:https://doi.org/10.1016/j.biortech.2012.04.022
Markou G, Chatzipavlidis I, Georgakakis D (2012) Carbohydrates production and bio-flocculation characteristics in cultures of Arthrospira (Spirulina) platensis: improvements through phosphorus limitation process. BioEnergy Res 5:915–925. doi:https://doi.org/10.1007/s12155-012-9205-3
Meixner K, Daffert C, Dalnodar D, Mrazova K, Hrubanova K, Krzyzanek V, Nebesarova J, Samek O, Šedrlová Z, Slaninova E (2022) Glycogen, poly (3-hydroxybutyrate) and pigment accumulation in three Synechocystis strains when exposed to a stepwise increasing salt stress. J Appl Phycol 34:1227–1241
Meuser JE, Boyd ES, Ananyev G, Karns D, Radakovits R, Narayana Murthy UM, Ghirardi ML, Dismukes GC, Peters JW, Posewitz MC (2011) Evolutionary significance of an algal gene encoding an [FeFe]-hydrogenase with F-domain homology and hydrogenase activity in Chlorella variabilis NC64A. Planta 234:829–843. doi:https://doi.org/10.1007/s00425-011-1431-y
Mills LA, McCormick AJ, Lea-Smith DJ (2020) Current knowledge and recent advances in understanding metabolism of the model cyanobacterium Synechocystis sp. PCC 6803. Biosci Rep 40. doi:https://doi.org/10.1042/bsr20193325
Monshupanee T, Incharoensakdi A (2014) Enhanced accumulation of glycogen, lipids and polyhydroxybutyrate under optimal nutrients and light intensities in the cyanobacterium Synechocystis sp. PCC 6803. J Appl Microbiol 116:830–838. doi:https://doi.org/10.1111/jam.12409
Monshupanee T, Nimdach P, Incharoensakdi A (2016) Two-stage (photoautotrophy and heterotrophy) cultivation enables efficient production of bioplastic poly-3-hydroxybutyrate in auto-sedimenting cyanobacterium. Sci Rep 6:37121. doi:https://doi.org/10.1038/srep37121
Muro-Pastor MI, Cutillas-Farray Á, Pérez-Rodríguez L, Pérez-Saavedra J, Vega-de Armas A, Paredes A, Robles-Rengel R, Florencio FJ (2020) CfrA, a novel carbon flow regulator, adapts carbon metabolism to nitrogen deficiency in cyanobacteria. Plant Physiol 184:1792–1810. doi:https://doi.org/10.1104/pp.20.00802
Nakamura Y, Takahashi J-I, Sakurai A, Inaba Y, Suzuki E, Nihei S, Fujiwara S, Tsuzuki M, Miyashita H, Ikemoto H, Kawachi M, Sekiguchi H, Kurano N (2005) Some cyanobacteria synthesize semi-amylopectin type α-polyglucans instead of glycogen. Plant Cell Physiol 46:539–545. doi:https://doi.org/10.1093/pcp/pci045
Orthwein T, Scholl J, Spät P, Lucius S, Koch M, Macek B, Hagemann M, Forchhammer K (2021) The novel PII-interactor PirC identifies phosphoglycerate mutase as key control point of carbon storage metabolism in cyanobacteria. Proc Natl Acad Sci U S A 118:e2019988118. doi:https://doi.org/10.1073/pnas.2019988118
Osanai T, Imamura S, Asayama M, Shirai M, Suzuki I, Murata N, Tanaka K (2006) Nitrogen induction of sugar catabolic gene expression in Synechocystis sp. PCC 6803. DNA Res 13:185–195. doi:https://doi.org/10.1093/dnares/dsl010
Osanai T, Oikawa A, Shirai T, Kuwahara A, Iijima H, Tanaka K, Ikeuchi M, Kondo A, Saito K, Hirai MY (2014) Capillary electrophoresis-mass spectrometry reveals the distribution of carbon metabolites during nitrogen starvation in Synechocystis sp. PCC 6803. Environ Microbiol 16:512–524. doi:https://doi.org/10.1111/1462-2920.12170
Panoff J-M, Priem B, Morvan H, Joset F (1988) Sulphated exopolysaccharides produced by two unicellular strains of cyanobacteria, Synechocystis PCC 6803 and 6714. Arch Microbiol 150:558–563
Patel VK, Sundaram S, Patel AK, Kalra A (2018) Characterization of seven species of cyanobacteria for high-quality biomass production. Arab J Sci Eng 43:109–121. doi:https://doi.org/10.1007/s13369-017-2666-0
Prasannan CB, Jaiswal D, Davis R, Wangikar PP (2018) An improved method for extraction of polar and charged metabolites from cyanobacteria. PLoS ONE 13:e0204273. doi:https://doi.org/10.1371/journal.pone.0204273
Rajeshwari KR, Rajashekhar M (2011) Biochemical composition of seven species of cyanobacteria isolated from different aquatic habitats of western ghats, Southern India. Braz Arch Biol Technol 54:849–857. doi:https://doi.org/10.1590/S1516-89132011000500001
Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J Gen Microbiol 111:1–61. doi:https://doi.org/10.1099/00221287-111-1-1
Schlebusch M, Forchhammer K (2010) Requirement of the nitrogen starvation-induced protein sll0783 for polyhydroxybutyrate accumulation in Synechocystis sp. strain PCC 6803. Appl Environ Microbiol 76:6101–6107. doi:https://doi.org/10.1128/AEM.00484-10
Scholl J, Dengler L, Bader L, Forchhammer K (2020) Phosphoenolpyruvate carboxylase from the cyanobacterium Synechocystis sp. PCC 6803 is under global metabolic control by PII signaling. Mol Microbiol 114:292–307. doi:https://doi.org/10.1111/mmi.14512
Singhon P, Phoraksa O, Incharoensakdi A, Monshupanee T (2021) Increased bioproduction of glycogen, lipids, and poly(3-hydroxybutyrate) under partial supply of nitrogen and phosphorus by photoautotrophic cyanobacterium Synechocystis sp. PCC 6803. J Appl Phycol 33:2833–2843. doi:https://doi.org/10.1007/s10811-021-02494-0
Sukkasam N, Incharoensakdi A, Monshupanee T (2021) Disruption of hydrogen gas synthesis enhances the cellular levels of NAD(P)H, glycogen, poly(3-hydroxybutyrate) and photosynthetic pigments under specific nutrient condition(s) in cyanobacterium Synechocystis sp. PCC 6803. Plant Cell Physiol 63:135–147. doi:https://doi.org/10.1093/pcp/pcab156
Toyoshima M, Tokumaru Y, Matsuda F, Shimizu H (2020) Assessment of protein content and phosphorylation level in Synechocystis sp. PCC 6803 under various growth conditions using quantitative phosphoproteomic analysis. Molecules 25:3582. doi:https://doi.org/10.3390/molecules25163582
Vonshak A, Guy R, Guy M (1988) The response of the filamentous cyanobacterium Spirulina platensis to salt stress. Arch Microbiol 150:417–420. doi:https://doi.org/10.1007/BF00422279
Welkie DG, Lee B-H, Sherman LA (2016) Altering the structure of carbohydrate storage granules in the cyanobacterium Synechocystis sp. strain PCC 6803 through branching-enzyme truncations. J Bacteriol 198:701–710. doi:https://doi.org/10.1128/JB.00830-15
Wishart DS, Feunang YD, Marcu A, Guo AC, Liang K, Vázquez-Fresno R, Sajed T, Johnson D, Li C, Karu N, Sayeeda Z, Lo E, Assempour N, Berjanskii M, Singhal S, Arndt D, Liang Y, Badran H, Grant J, Serra-Cayuela A, Liu Y, Mandal R, Neveu V, Pon A, Knox C, Wilson M, Manach C, Scalbert A (2018) HMDB 4.0: the human metabolome database for 2018. Nucleic Acids Res 46:D608–D617. doi:https://doi.org/10.1093/nar/gkx1089
Wishart DS, Tzur D, Knox C, Eisner R, Guo AC, Young N, Cheng D, Jewell K, Arndt D, Sawhney S, Fung C, Nikolai L, Lewis M, Coutouly M-A, Forsythe I, Tang P, Shrivastava S, Jeroncic K, Stothard P, Amegbey G, Block D, Hau DD, Wagner J, Miniaci J, Clements M, Gebremedhin M, Guo N, Zhang Y, Duggan GE, Macinnis GD, Weljie AM, Dowlatabadi R, Bamforth F, Clive D, Greiner R, Li L, Marrie T, Sykes BD, Vogel HJ, Querengesser L (2007) HMDB: the human metabolome database. Nucleic Acids Res 35:D521–D526. doi:https://doi.org/10.1093/nar/gkl923
Zhao M-X, Jiang Y-L, Xu B-Y, Chen Y, Zhang C-C, Zhou C-Z (2010) Crystal structure of the cyanobacterial signal transduction protein PII in complex with PipX. J Mol Biol 402:552–559. doi:https://doi.org/10.1016/j.jmb.2010.08.006
Acknowledgements
The authors thank TISTR and Dr. Aparat Mahakhant for cyanobacterial TISTR strains and Dr. Robert Butcher for critical proofreading.
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This project is funded by National Research Council of Thailand (NRCT): NRCT5-RSA63001-21 (to Tanakarn Monshupanee). Panutchaya Pichaiyotinkul was funded by CU Graduate School Thesis Grant GCUGR1225642068M and by a Chulalongkorn University Graduate Scholarship to Commemorate the 72nd Anniversary of His Majesty King Bhumibol Adulyadej.
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Panutchaya Pichaiyotinkul and Tanakarn Monshupanee developed the concept and designed the experiments with the input from Aran Incharoensakdi. Panutchaya Pichaiyotinkul and Tanakarn Monshupanee conducted the experiments, analyzed the data and wrote the manuscript. Nathanich Ruankaew conducted the experiment under the deprivation of both nitrogen and phosphorus. All authors reviewed the manuscript. Tanakarn Monshupanee is responsible for funding acquisition.
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Pichaiyotinkul, P., Ruankaew, N., Incharoensakdi, A. et al. Enhanced polyglucan contents in divergent cyanobacteria under nutrient-deprived photoautotrophy: transcriptional and metabolic changes in response to increased glycogen accumulation in nitrogen-deprived Synechocystis sp. PCC 6803. World J Microbiol Biotechnol 39, 27 (2023). https://doi.org/10.1007/s11274-022-03476-1
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DOI: https://doi.org/10.1007/s11274-022-03476-1