{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,24]],"date-time":"2026-06-24T14:35:33Z","timestamp":1782311733271,"version":"3.54.5"},"reference-count":41,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T00:00:00Z","timestamp":1764028800000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T00:00:00Z","timestamp":1764028800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100000266","name":"Engineering and Physical Sciences Research Council","doi-asserted-by":"publisher","award":["EP\/S01778X\/1"],"award-info":[{"award-number":["EP\/S01778X\/1"]}],"id":[{"id":"10.13039\/501100000266","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Biotechnol. Biofuels Bioprod."],"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Genetic and metabolic engineering of\n                    <jats:italic>Synechocystis<\/jats:italic>\n                    sp. PCC 6803 has given rise to strains that produce a variety of bio-chemicals. Despite these successes, improvements in productivity metrics are required to achieve economically viable bio-production of carbon dioxide-derived compounds. Previously, environmental factors have been leveraged to increase product yields. Here, we optimise multiple environmental factors simultaneously using Design of Experiments (DOE) principles and find conditions that maximise L-lactate production. Light intensity, glycerol concentration, and light\u2013dark cycle were found to be significant factors. Optimising these conditions resulted in a 6.3- and 7.4-fold increase in titre and yield. The results detailed here could have implications for metabolic engineering of, and bioprocesses using,\n                    <jats:italic>Synechocystis<\/jats:italic>\n                    sp. PCC 6803. In the future, DOE-mediated optimisation of environmental conditions could effectively maximise product titres from different production strains, or the enhanced conditions described here could be directly implemented in other metabolic engineering projects.\n                  <\/jats:p>","DOI":"10.1186\/s13068-025-02718-2","type":"journal-article","created":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T09:13:04Z","timestamp":1764061984000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Optimising environmental factors for maximal lactate productivity in Synechocystis sp. PCC 6803 through a design of experiments approach"],"prefix":"10.1186","volume":"18","author":[{"given":"Matthew","family":"Faulkner","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fraser","family":"Andrews","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nigel S.","family":"Scrutton","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,11,25]]},"reference":[{"key":"2718_CR1","doi-asserted-by":"publisher","DOI":"10.3390\/life6040042","volume":"6","author":"L Al-Haj","year":"2016","unstructured":"Al-Haj L, Lui YT, Abed RMM, Gomaa MA, Purton S. Cyanobacteria as chassis for industrial biotechnology: progress and prospects. Life. 2016;6:42.","journal-title":"Life"},{"key":"2718_CR2","doi-asserted-by":"publisher","first-page":"5481","DOI":"10.1007\/s11033-023-08456-8","volume":"50","author":"J Cheng","year":"2023","unstructured":"Cheng J, Zhang K, Hou Y. The current situations and limitations of genetic engineering in cyanobacteria: a mini review. Mol Biol Rep. 2023;50:5481\u20137.","journal-title":"Mol Biol Rep"},{"key":"2718_CR3","doi-asserted-by":"publisher","DOI":"10.1016\/j.biotechadv.2020.107517","volume":"40","author":"A Srivastava","year":"2020","unstructured":"Srivastava A, Summers ML, Sobotka R. Cyanobacterial sigma factors: current and future applications for biotechnological advances. Biotechnol Adv. 2020;40:107517.","journal-title":"Biotechnol Adv"},{"key":"2718_CR4","doi-asserted-by":"publisher","first-page":"240","DOI":"10.1186\/s13068-021-02091-w","volume":"14","author":"F Andrews","year":"2021","unstructured":"Andrews F, Faulkner M, Toogood HS, Scrutton NS. Combinatorial use of environmental stresses and genetic engineering to increase ethanol titres in cyanobacteria. Biotechnol Biofuels. 2021;14:240.","journal-title":"Biotechnol Biofuels"},{"key":"2718_CR5","doi-asserted-by":"publisher","first-page":"101","DOI":"10.3390\/metabo3010101","volume":"3","author":"W Hauf","year":"2013","unstructured":"Hauf W, et al. Metabolic changes in Synechocystis PCC6803 upon nitrogen-starvation: excess NADPH sustains polyhydroxybutyrate accumulation. Metabolites. 2013;3:101\u201318.","journal-title":"Metabolites"},{"key":"2718_CR6","doi-asserted-by":"publisher","first-page":"537","DOI":"10.1093\/pcp\/pcx047","volume":"58","author":"T Nakajima","year":"2017","unstructured":"Nakajima T, Yoshikawa K, Toya Y, Matsuda F, Shimizu H. Metabolic flux analysis of the Synechocystis sp. PCC 6803 \u0394nrtABCD mutant reveals a mechanism for metabolic adaptation to nitrogen-limited conditions. Plant Cell Physiol. 2017;58:537\u201345.","journal-title":"Plant Cell Physiol"},{"key":"2718_CR7","doi-asserted-by":"publisher","DOI":"10.1186\/s13068-024-02589-z","volume":"17","author":"M Faulkner","year":"2024","unstructured":"Faulkner M, Andrews F, Scrutton N. Improving productivity of citramalate from CO2 by Synechocystis sp. PCC 6803 through design of experiment. Biotechnol Biofuels Bioprod. 2024;17:143.","journal-title":"Biotechnol Biofuels Bioprod"},{"key":"2718_CR8","doi-asserted-by":"publisher","DOI":"10.3389\/fpls.2020.586543","volume":"11","author":"M Canonico","year":"2020","unstructured":"Canonico M, Konert G, Ka\u0148a R. Plasticity of cyanobacterial thylakoid microdomains under variable light conditions. Front Plant Sci. 2020;11:586543.","journal-title":"Front Plant Sci"},{"key":"2718_CR9","doi-asserted-by":"publisher","first-page":"2263","DOI":"10.1021\/acssynbio.9b00289","volume":"8","author":"W Du","year":"2019","unstructured":"Du W, et al. Exploiting day- and night-time metabolism of Synechocystis sp. PCC 6803 for fitness-coupled fumarate production around the clock. ACS Synth Biol. 2019;8:2263\u20139.","journal-title":"ACS Synth Biol"},{"key":"2718_CR10","doi-asserted-by":"publisher","first-page":"4","DOI":"10.1186\/s13068-022-02237-4","volume":"16","author":"GD Barone","year":"2023","unstructured":"Barone GD, et al. Towards the rate limit of heterologous biotechnological reactions in recombinant cyanobacteria. Biotechnol Biofuels Bioprod. 2023;16:4.","journal-title":"Biotechnol Biofuels Bioprod"},{"key":"2718_CR11","doi-asserted-by":"publisher","first-page":"667","DOI":"10.1016\/j.celrep.2018.03.083","volume":"23","author":"M Cano","year":"2018","unstructured":"Cano M, et al. Glycogen synthesis and metabolite overflow contribute to energy balancing in cyanobacteria. Cell Rep. 2018;23:667\u201372.","journal-title":"Cell Rep"},{"key":"2718_CR12","doi-asserted-by":"publisher","first-page":"3063","DOI":"10.1007\/s00158-021-02977-1","volume":"64","author":"R Le Riche","year":"2021","unstructured":"Le Riche R, Picheny V. Revisiting Bayesian optimization in the light of the COCO benchmark. Struct Multidiscip Optim. 2021;64:3063\u201387.","journal-title":"Struct Multidiscip Optim"},{"key":"2718_CR13","doi-asserted-by":"publisher","DOI":"10.1007\/978-94-009-0909-0","volume-title":"Bayesian approach to global optimization theory and applications","author":"J Mockus","year":"1989","unstructured":"Mockus J. Global optimization and the bayesian approach. In: Mockus J, editor. Bayesian approach to global optimization theory and applications. Dordrecht: Springer; 1989."},{"key":"2718_CR14","volume-title":"Current developments in biotechnology and bioengineering","author":"R Balakrishnan","year":"2022","unstructured":"Balakrishnan R, Mohan N, Sivaprakasam S. Chapter 11 - Application of design of experiments in bioprocessing: process analysis, optimization, and reliability. In: Sirohi R, Pandey A, Taherzadeh MJ, Larroche C, editors. Current developments in biotechnology and bioengineering. Amsterdam: Elsevier; 2022."},{"key":"2718_CR15","doi-asserted-by":"publisher","first-page":"368","DOI":"10.3109\/07388551.2014.973014","volume":"36","author":"T Keskin G\u00fcndo\u011fdu","year":"2016","unstructured":"Keskin G\u00fcndo\u011fdu T, Deniz \u0130, \u00c7al\u0131\u015fkan G, \u015eahin ES, Azbar N. Experimental design methods for bioengineering applications. Crit Rev Biotechnol. 2016;36:368\u201388.","journal-title":"Crit Rev Biotechnol"},{"key":"2718_CR16","doi-asserted-by":"publisher","DOI":"10.1016\/j.biortech.2023.129677","volume":"387","author":"I Yadav","year":"2023","unstructured":"Yadav I, et al. Enhancement of isoprene production in engineered Synechococcus elongatus UTEX 2973 by metabolic pathway inhibition and machine learning-based optimization strategy. Bioresour Technol. 2023;387:129677.","journal-title":"Bioresour Technol"},{"key":"2718_CR17","doi-asserted-by":"publisher","first-page":"4312","DOI":"10.3390\/ma15124312","volume":"15","author":"L Ranakoti","year":"2022","unstructured":"Ranakoti L, et al. Critical review on polylactic acid: properties, structure, processing, biocomposites, and nanocomposites. Materials. 2022;15:4312.","journal-title":"Materials"},{"key":"2718_CR18","doi-asserted-by":"publisher","first-page":"2318","DOI":"10.3390\/microorganisms10122318","volume":"10","author":"E \u017byma\u0144czyk-Duda","year":"2022","unstructured":"\u017byma\u0144czyk-Duda E, Samson SO, Brzezi\u0144ska-Rodak M, Klimek-Ochab M. Versatile applications of cyanobacteria in biotechnology. Microorganisms. 2022;10:2318.","journal-title":"Microorganisms"},{"key":"2718_CR19","doi-asserted-by":"publisher","first-page":"3032","DOI":"10.1099\/mic.0.062950-0","volume":"158","author":"M Gr\u00fcndel","year":"2012","unstructured":"Gr\u00fcndel M, Scheunemann R, Lockau W, Zilliges Y. Impaired glycogen synthesis causes metabolic overflow reactions and affects stress responses in the cyanobacterium Synechocystis sp. PCC 6803. Microbiology. 2012;158:3032\u201343.","journal-title":"Microbiology"},{"key":"2718_CR20","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1016\/j.jbiotec.2016.09.016","volume":"239","author":"K Namakoshi","year":"2016","unstructured":"Namakoshi K, Nakajima T, Yoshikawa K, Toya Y, Shimizu H. Combinatorial deletions of glgC and phaCE enhance ethanol production in Synechocystis sp. PCC 6803. J Biotechnol. 2016;239:13\u20139.","journal-title":"J Biotechnol"},{"key":"2718_CR21","doi-asserted-by":"publisher","first-page":"202","DOI":"10.1006\/mben.2002.0226","volume":"4","author":"C Yang","year":"2002","unstructured":"Yang C, Hua Q, Shimizu K. Metabolic flux analysis in Synechocystis using isotope distribution from 13C-labeled glucose. Metab Eng. 2002;4:202\u201316.","journal-title":"Metab Eng"},{"key":"2718_CR22","doi-asserted-by":"publisher","first-page":"E580","DOI":"10.1073\/pnas.1613078114","volume":"114","author":"S Diamond","year":"2017","unstructured":"Diamond S, et al. Redox crisis underlies conditional light-dark lethality in cyanobacterial mutants that lack the circadian regulator, RpaA. Proc Natl Acad Sci USA. 2017;114:E580\u20139.","journal-title":"Proc Natl Acad Sci USA"},{"key":"2718_CR23","doi-asserted-by":"publisher","first-page":"E1916","DOI":"10.1073\/pnas.1504576112","volume":"112","author":"S Diamond","year":"2015","unstructured":"Diamond S, Jun D, Rubin BE, Golden SS. The circadian oscillator in Synechococcus elongatus controls metabolite partitioning during diurnal growth. Proc Natl Acad Sci U S A. 2015;112:E1916\u201325.","journal-title":"Proc Natl Acad Sci U S A"},{"key":"2718_CR24","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41540-023-00307-3","volume":"9","author":"A Kugler","year":"2023","unstructured":"Kugler A, Stensj\u00f6 K. Optimal energy and redox metabolism in the cyanobacterium Synechocystis sp. PCC 6803. npj Syst Biol Appl. 2023;9:1\u201313.","journal-title":"npj Syst Biol Appl"},{"key":"2718_CR25","doi-asserted-by":"publisher","first-page":"194","DOI":"10.1111\/nph.14353","volume":"214","author":"J K\u00e4m\u00e4r\u00e4inen","year":"2017","unstructured":"K\u00e4m\u00e4r\u00e4inen J, et al. Pyridine nucleotide transhydrogenase PntAB is essential for optimal growth and photosynthetic integrity under low-light mixotrophic conditions in Synechocystis sp. PCC 6803. New Phytol. 2017;214:194\u2013204.","journal-title":"New Phytol"},{"key":"2718_CR26","doi-asserted-by":"publisher","first-page":"270","DOI":"10.1093\/gbe\/evy275","volume":"11","author":"SE Will","year":"2019","unstructured":"Will SE, et al. Day and night: metabolic profiles and evolutionary relationships of six axenic non-marine cyanobacteria. Genome Biol Evol. 2019;11:270\u201394.","journal-title":"Genome Biol Evol"},{"key":"2718_CR27","doi-asserted-by":"publisher","first-page":"130","DOI":"10.1016\/j.meteno.2016.04.003","volume":"3","author":"T Hasunuma","year":"2016","unstructured":"Hasunuma T, Matsuda M, Kondo A. Improved sugar-free succinate production by Synechocystis sp. PCC 6803 following identification of the limiting steps in glycogen catabolism. Metab Eng Commun. 2016;3:130\u201341.","journal-title":"Metab Eng Commun"},{"key":"2718_CR28","doi-asserted-by":"publisher","first-page":"275","DOI":"10.1007\/BF02178570","volume":"8","author":"R De Philippis","year":"1996","unstructured":"De Philippis R, Sili C, Vincenzini M. Response of an exopolysaccharide-producing heterocystous cyanobacterium to changes in metabolic carbon flux. J Appl Phycol. 1996;8:275\u201381.","journal-title":"J Appl Phycol"},{"key":"2718_CR29","doi-asserted-by":"publisher","DOI":"10.1186\/s13068-020-1656-8","volume":"13","author":"JK Gupta","year":"2020","unstructured":"Gupta JK, Rai P, Jain KK, Srivastava S. Overexpression of bicarbonate transporters in the marine cyanobacterium Synechococcus sp. PCC 7002 increases growth rate and glycogen accumulation. Biotechnol Biofuels. 2020;13:17.","journal-title":"Biotechnol Biofuels"},{"key":"2718_CR30","doi-asserted-by":"publisher","first-page":"76","DOI":"10.1016\/j.ymben.2015.03.002","volume":"29","author":"NA Kamennaya","year":"2015","unstructured":"Kamennaya NA, et al. Installing extra bicarbonate transporters in the cyanobacterium Synechocystis sp. PCC6803 enhances biomass production. Metab Eng. 2015;29:76\u201385.","journal-title":"Metab Eng"},{"key":"2718_CR31","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0056887","volume":"8","author":"J St\u00f6ckel","year":"2013","unstructured":"St\u00f6ckel J, Elvitigala TR, Liberton M, Pakrasi HB. Carbon availability affects diurnally controlled processes and cell morphology of Cyanothece 51142. PLoS ONE. 2013;8:e56887.","journal-title":"PLoS ONE"},{"key":"2718_CR32","doi-asserted-by":"publisher","first-page":"1333","DOI":"10.1007\/s00253-015-7024-0","volume":"100","author":"E Touloupakis","year":"2016","unstructured":"Touloupakis E, Cicchi B, Benavides AMS, Torzillo G. Effect of high pH on growth of Synechocystis sp. PCC 6803 cultures and their contamination by golden algae (Poterioochromonas sp.). Appl Microbiol Biotechnol. 2016;100:1333\u201341.","journal-title":"Appl Microbiol Biotechnol"},{"key":"2718_CR33","doi-asserted-by":"publisher","first-page":"121","DOI":"10.1016\/j.coche.2016.09.008","volume":"14","author":"JM Burg","year":"2016","unstructured":"Burg JM, et al. Large-scale bioprocess competitiveness: the potential of dynamic metabolic control in two-stage fermentations. Curr Opin Chem Eng. 2016;14:121\u201336.","journal-title":"Curr Opin Chem Eng"},{"key":"2718_CR34","doi-asserted-by":"publisher","DOI":"10.1038\/s44222-024-00225-x","author":"K Shabestary","year":"2024","unstructured":"Shabestary K, et al. Design of microbial catalysts for two-stage processes. Nat Rev Bioeng. 2024. https:\/\/doi.org\/10.1038\/s44222-024-00225-x.","journal-title":"Nat Rev Bioeng"},{"key":"2718_CR35","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1016\/j.ymben.2021.09.010","volume":"68","author":"K Shabestary","year":"2021","unstructured":"Shabestary K, et al. Cycling between growth and production phases increases cyanobacteria bioproduction of lactate. Metab Eng. 2021;68:131\u201341.","journal-title":"Metab Eng"},{"key":"2718_CR36","doi-asserted-by":"publisher","first-page":"1669","DOI":"10.1021\/acssynbio.8b00056","volume":"7","author":"K Shabestary","year":"2018","unstructured":"Shabestary K, et al. Targeted repression of essential genes to arrest growth and increase carbon partitioning and biofuel titers in cyanobacteria. ACS Synth Biol. 2018;7:1669\u201375.","journal-title":"ACS Synth Biol"},{"key":"2718_CR37","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1099\/00221287-111-1-1","volume":"111","author":"R Rippka","year":"1979","unstructured":"Rippka R, Deruelles J, Waterbury JB, Herdman M, Stanier RY. Generic assignments, strain histories and properties of pure cultures of cyanobacteria. Microbiology. 1979;111:1\u201361.","journal-title":"Microbiology"},{"key":"2718_CR38","doi-asserted-by":"publisher","first-page":"26","DOI":"10.1186\/s13062-021-00316-4","volume":"16","author":"S Kl\u00e4hn","year":"2021","unstructured":"Kl\u00e4hn S, et al. Integrative analysis of the salt stress response in cyanobacteria. Biol Direct. 2021;16:26.","journal-title":"Biol Direct"},{"key":"2718_CR39","doi-asserted-by":"publisher","DOI":"10.1186\/1754-6834-7-99","volume":"7","author":"SA Angermayr","year":"2014","unstructured":"Angermayr SA, et al. Exploring metabolic engineering design principles for the photosynthetic production of lactic acid by Synechocystis sp. PCC6803. Biotechnol Biofuels. 2014;7:99.","journal-title":"Biotechnol Biofuels"},{"key":"2718_CR40","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1021\/acssynbio.6b00239","volume":"6","author":"M Kanno","year":"2017","unstructured":"Kanno M, Atsumi S. Engineering an obligate photoautotrophic cyanobacterium to utilize glycerol for growth and chemical production. ACS Synth Biol. 2017;6:69\u201375.","journal-title":"ACS Synth Biol"},{"key":"2718_CR41","doi-asserted-by":"publisher","first-page":"11169","DOI":"10.1021\/jp4013152","volume":"117","author":"SA Angermayr","year":"2013","unstructured":"Angermayr SA, Hellingwerf KJ. On the use of metabolic control analysis in the optimization of cyanobacterial biosolar cell factories. J Phys Chem B. 2013;117:11169\u201375.","journal-title":"J Phys Chem B"}],"container-title":["Biotechnology for Biofuels and Bioproducts"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13068-025-02718-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/article\/10.1186\/s13068-025-02718-2\/fulltext.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/link.springer.com\/content\/pdf\/10.1186\/s13068-025-02718-2.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,25]],"date-time":"2025-11-25T09:13:05Z","timestamp":1764061985000},"score":1,"resource":{"primary":{"URL":"https:\/\/biotechnologyforbiofuels.biomedcentral.com\/articles\/10.1186\/s13068-025-02718-2"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,11,25]]},"references-count":41,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2025,12]]}},"alternative-id":["2718"],"URL":"https:\/\/doi.org\/10.1186\/s13068-025-02718-2","relation":{},"ISSN":["2731-3654"],"issn-type":[{"value":"2731-3654","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,11,25]]},"assertion":[{"value":"6 July 2025","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"6 November 2025","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"25 November 2025","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Declarations"}},{"value":"The authors declare no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"118"}}