[{"article_number":"e10490","title":"Growth‐mediated negative feedback shapes quantitative antibiotic response","author":[{"first_name":"Andreas","id":"4677C796-F248-11E8-B48F-1D18A9856A87","last_name":"Angermayr","orcid":"0000-0001-8619-2223","full_name":"Angermayr, Andreas"},{"full_name":"Pang, Tin Yau","last_name":"Pang","first_name":"Tin Yau"},{"first_name":"Guillaume","full_name":"Chevereau, Guillaume","last_name":"Chevereau"},{"first_name":"Karin","id":"39B66846-F248-11E8-B48F-1D18A9856A87","full_name":"Mitosch, Karin","last_name":"Mitosch"},{"first_name":"Martin J","full_name":"Lercher, Martin J","last_name":"Lercher"},{"full_name":"Bollenbach, Mark Tobias","orcid":"0000-0003-4398-476X","last_name":"Bollenbach","first_name":"Mark Tobias","id":"3E6DB97A-F248-11E8-B48F-1D18A9856A87"}],"external_id":{"isi":["000856482800001"]},"article_processing_charge":"No","user_id":"4359f0d1-fa6c-11eb-b949-802e58b17ae8","citation":{"chicago":"Angermayr, Andreas, Tin Yau Pang, Guillaume Chevereau, Karin Mitosch, Martin J Lercher, and Mark Tobias Bollenbach. “Growth‐mediated Negative Feedback Shapes Quantitative Antibiotic Response.” Molecular Systems Biology. Embo Press, 2022. https://doi.org/10.15252/msb.202110490.","ista":"Angermayr A, Pang TY, Chevereau G, Mitosch K, Lercher MJ, Bollenbach MT. 2022. Growth‐mediated negative feedback shapes quantitative antibiotic response. Molecular Systems Biology. 18(9), e10490.","mla":"Angermayr, Andreas, et al. “Growth‐mediated Negative Feedback Shapes Quantitative Antibiotic Response.” Molecular Systems Biology, vol. 18, no. 9, e10490, Embo Press, 2022, doi:10.15252/msb.202110490.","ama":"Angermayr A, Pang TY, Chevereau G, Mitosch K, Lercher MJ, Bollenbach MT. Growth‐mediated negative feedback shapes quantitative antibiotic response. Molecular Systems Biology. 2022;18(9). doi:10.15252/msb.202110490","apa":"Angermayr, A., Pang, T. Y., Chevereau, G., Mitosch, K., Lercher, M. J., & Bollenbach, M. T. (2022). Growth‐mediated negative feedback shapes quantitative antibiotic response. Molecular Systems Biology. Embo Press. https://doi.org/10.15252/msb.202110490","short":"A. Angermayr, T.Y. Pang, G. Chevereau, K. Mitosch, M.J. Lercher, M.T. Bollenbach, Molecular Systems Biology 18 (2022).","ieee":"A. Angermayr, T. Y. Pang, G. Chevereau, K. Mitosch, M. J. Lercher, and M. T. Bollenbach, “Growth‐mediated negative feedback shapes quantitative antibiotic response,” Molecular Systems Biology, vol. 18, no. 9. Embo Press, 2022."},"quality_controlled":"1","publisher":"Embo Press","oa":1,"acknowledgement":"This work was in part supported by Human Frontier Science Program GrantRGP0042/2013, Marie Curie Career Integration Grant303507, AustrianScience Fund (FWF) Grant P27201-B22, and German Research Foundation(DFG) Collaborative Research Center (SFB)1310to TB. SAA was supportedby the European Union’s Horizon2020Research and Innovation Programunder the Marie Skłodowska-Curie Grant agreement No707352. We wouldlike to thank the Bollenbach group for regular fruitful discussions. We areparticularly thankful for the technical assistance of Booshini Fernando andfor discussions of the theoretical aspects with Gerrit Ansmann. We areindebted to Bor Kavˇciˇc for invaluable advice, help with setting up theluciferase-based growth monitoring system, and for sharing plasmids. Weacknowledge the IST Austria Miba Machine Shop for their support inbuilding a housing for the stacker of the plate reader, which enabled thehigh-throughput luciferase-based experiments. We are grateful to RosalindAllen, Bor Kavˇciˇc and Dor Russ for feedback on the manuscript. Open Accessfunding enabled and organized by Projekt DEAL.","date_published":"2022-09-01T00:00:00Z","doi":"10.15252/msb.202110490","date_created":"2023-01-16T09:58:34Z","day":"01","publication":"Molecular Systems Biology","has_accepted_license":"1","isi":1,"year":"2022","status":"public","keyword":["Applied Mathematics","Computational Theory and Mathematics","General Agricultural and Biological Sciences","General Immunology and Microbiology","General Biochemistry","Genetics and Molecular Biology","Information Systems"],"article_type":"original","type":"journal_article","tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","short":"CC BY (4.0)"},"_id":"12261","file_date_updated":"2023-01-30T09:49:55Z","department":[{"_id":"ToBo"}],"ddc":["570"],"date_updated":"2023-08-04T09:51:49Z","month":"09","intvolume":" 18","scopus_import":"1","oa_version":"Published Version","acknowledged_ssus":[{"_id":"M-Shop"}],"abstract":[{"lang":"eng","text":"Dose–response relationships are a general concept for quantitatively describing biological systems across multiple scales, from the molecular to the whole-cell level. A clinically relevant example is the bacterial growth response to antibiotics, which is routinely characterized by dose–response curves. The shape of the dose–response curve varies drastically between antibiotics and plays a key role in treatment, drug interactions, and resistance evolution. However, the mechanisms shaping the dose–response curve remain largely unclear. Here, we show in Escherichia coli that the distinctively shallow dose–response curve of the antibiotic trimethoprim is caused by a negative growth-mediated feedback loop: Trimethoprim slows growth, which in turn weakens the effect of this antibiotic. At the molecular level, this feedback is caused by the upregulation of the drug target dihydrofolate reductase (FolA/DHFR). We show that this upregulation is not a specific response to trimethoprim but follows a universal trend line that depends primarily on the growth rate, irrespective of its cause. Rewiring the feedback loop alters the dose–response curve in a predictable manner, which we corroborate using a mathematical model of cellular resource allocation and growth. Our results indicate that growth-mediated feedback loops may shape drug responses more generally and could be exploited to design evolutionary traps that enable selection against drug resistance."}],"issue":"9","volume":18,"license":"https://creativecommons.org/licenses/by/4.0/","file":[{"access_level":"open_access","relation":"main_file","content_type":"application/pdf","file_id":"12446","checksum":"8b1d8f5ea20c8408acf466435fb6ae01","success":1,"creator":"dernst","date_updated":"2023-01-30T09:49:55Z","file_size":1098812,"date_created":"2023-01-30T09:49:55Z","file_name":"2022_MolecularSystemsBio_Angermayr.pdf"}],"language":[{"iso":"eng"}],"publication_identifier":{"eissn":["1744-4292"]},"publication_status":"published"},{"date_updated":"2021-01-12T08:01:21Z","department":[{"_id":"ToBo"}],"_id":"520","status":"public","article_type":"letter_note","type":"journal_article","language":[{"iso":"eng"}],"publication_status":"published","publication_identifier":{"issn":["21615063"]},"volume":6,"issue":"3","oa_version":"None","pmid":1,"abstract":[{"lang":"eng","text":"Cyanobacteria are mostly engineered to be sustainable cell-factories by genetic manipulations alone. Here, by modulating the concentration of allosteric effectors, we focus on increasing product formation without further burdening the cells with increased expression of enzymes. Resorting to a novel 96-well microplate cultivation system for cyanobacteria, and using lactate-producing strains of Synechocystis PCC6803 expressing different l-lactate dehydrogenases (LDH), we titrated the effect of 2,5-anhydro-mannitol supplementation. The latter acts in cells as a nonmetabolizable analogue of fructose 1,6-bisphosphate, a known allosteric regulator of one of the tested LDHs. In this strain (SAA023), we achieved over 2-fold increase of lactate productivity. Furthermore, we observed that as carbon is increasingly deviated during growth toward product formation, there is an increased fixation rate in the population of spontaneous mutants harboring an impaired production pathway. This is a challenge in the development of green cell factories, which may be countered by the incorporation in biotechnological processes of strategies such as the one pioneered here."}],"intvolume":" 6","month":"03","scopus_import":1,"user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","citation":{"ista":"Du W, Angermayr A, Jongbloets J, Molenaar D, Bachmann H, Hellingwerf K, Branco Dos Santos F. 2017. Nonhierarchical flux regulation exposes the fitness burden associated with lactate production in Synechocystis sp. PCC6803. ACS Synthetic Biology. 6(3), 395–401.","chicago":"Du, Wei, Andreas Angermayr, Joeri Jongbloets, Douwe Molenaar, Herwig Bachmann, Klaas Hellingwerf, and Filipe Branco Dos Santos. “Nonhierarchical Flux Regulation Exposes the Fitness Burden Associated with Lactate Production in Synechocystis Sp. PCC6803.” ACS Synthetic Biology. American Chemical Society, 2017. https://doi.org/10.1021/acssynbio.6b00235.","apa":"Du, W., Angermayr, A., Jongbloets, J., Molenaar, D., Bachmann, H., Hellingwerf, K., & Branco Dos Santos, F. (2017). Nonhierarchical flux regulation exposes the fitness burden associated with lactate production in Synechocystis sp. PCC6803. ACS Synthetic Biology. American Chemical Society. https://doi.org/10.1021/acssynbio.6b00235","ama":"Du W, Angermayr A, Jongbloets J, et al. Nonhierarchical flux regulation exposes the fitness burden associated with lactate production in Synechocystis sp. PCC6803. ACS Synthetic Biology. 2017;6(3):395-401. doi:10.1021/acssynbio.6b00235","ieee":"W. Du et al., “Nonhierarchical flux regulation exposes the fitness burden associated with lactate production in Synechocystis sp. PCC6803,” ACS Synthetic Biology, vol. 6, no. 3. American Chemical Society, pp. 395–401, 2017.","short":"W. Du, A. Angermayr, J. Jongbloets, D. Molenaar, H. Bachmann, K. Hellingwerf, F. Branco Dos Santos, ACS Synthetic Biology 6 (2017) 395–401.","mla":"Du, Wei, et al. “Nonhierarchical Flux Regulation Exposes the Fitness Burden Associated with Lactate Production in Synechocystis Sp. PCC6803.” ACS Synthetic Biology, vol. 6, no. 3, American Chemical Society, 2017, pp. 395–401, doi:10.1021/acssynbio.6b00235."},"title":"Nonhierarchical flux regulation exposes the fitness burden associated with lactate production in Synechocystis sp. PCC6803","external_id":{"pmid":["27936615"]},"publist_id":"7298","author":[{"first_name":"Wei","full_name":"Du, Wei","last_name":"Du"},{"orcid":"0000-0001-8619-2223","full_name":"Angermayr, Andreas","last_name":"Angermayr","first_name":"Andreas","id":"4677C796-F248-11E8-B48F-1D18A9856A87"},{"full_name":"Jongbloets, Joeri","last_name":"Jongbloets","first_name":"Joeri"},{"first_name":"Douwe","last_name":"Molenaar","full_name":"Molenaar, Douwe"},{"first_name":"Herwig","last_name":"Bachmann","full_name":"Bachmann, Herwig"},{"last_name":"Hellingwerf","full_name":"Hellingwerf, Klaas","first_name":"Klaas"},{"full_name":"Branco Dos Santos, Filipe","last_name":"Branco Dos Santos","first_name":"Filipe"}],"publication":"ACS Synthetic Biology","day":"17","year":"2017","date_created":"2018-12-11T11:46:56Z","date_published":"2017-03-17T00:00:00Z","doi":"10.1021/acssynbio.6b00235","page":"395 - 401","quality_controlled":"1","publisher":"American Chemical Society"},{"day":"23","publication":"Microbial Cell Factories","has_accepted_license":"1","isi":1,"year":"2017","date_published":"2017-02-23T00:00:00Z","doi":"10.1186/s12934-017-0645-5","date_created":"2018-12-11T11:49:56Z","quality_controlled":"1","publisher":"BioMed Central","oa":1,"user_id":"c635000d-4b10-11ee-a964-aac5a93f6ac1","citation":{"ista":"Veetil V, Angermayr A, Hellingwerf K. 2017. Ethylene production with engineered Synechocystis sp PCC 6803 strains. Microbial Cell Factories. 16(1), 34.","chicago":"Veetil, Vinod, Andreas Angermayr, and Klaas Hellingwerf. “Ethylene Production with Engineered Synechocystis Sp PCC 6803 Strains.” Microbial Cell Factories. BioMed Central, 2017. https://doi.org/10.1186/s12934-017-0645-5.","ama":"Veetil V, Angermayr A, Hellingwerf K. Ethylene production with engineered Synechocystis sp PCC 6803 strains. Microbial Cell Factories. 2017;16(1). doi:10.1186/s12934-017-0645-5","apa":"Veetil, V., Angermayr, A., & Hellingwerf, K. (2017). Ethylene production with engineered Synechocystis sp PCC 6803 strains. Microbial Cell Factories. BioMed Central. https://doi.org/10.1186/s12934-017-0645-5","short":"V. Veetil, A. Angermayr, K. Hellingwerf, Microbial Cell Factories 16 (2017).","ieee":"V. Veetil, A. Angermayr, and K. Hellingwerf, “Ethylene production with engineered Synechocystis sp PCC 6803 strains,” Microbial Cell Factories, vol. 16, no. 1. BioMed Central, 2017.","mla":"Veetil, Vinod, et al. “Ethylene Production with Engineered Synechocystis Sp PCC 6803 Strains.” Microbial Cell Factories, vol. 16, no. 1, 34, BioMed Central, 2017, doi:10.1186/s12934-017-0645-5."},"title":"Ethylene production with engineered Synechocystis sp PCC 6803 strains","author":[{"first_name":"Vinod","full_name":"Veetil, Vinod","last_name":"Veetil"},{"orcid":"0000-0001-8619-2223","full_name":"Angermayr, Andreas","last_name":"Angermayr","first_name":"Andreas","id":"4677C796-F248-11E8-B48F-1D18A9856A87"},{"first_name":"Klaas","full_name":"Hellingwerf, Klaas","last_name":"Hellingwerf"}],"publist_id":"6325","external_id":{"pmid":["28231787"],"isi":["000397733000001"]},"article_processing_charge":"No","article_number":"34","file":[{"file_id":"5240","content_type":"application/pdf","relation":"main_file","access_level":"open_access","file_name":"IST-2017-792-v1+1_s12934-017-0645-5.pdf","date_created":"2018-12-12T10:16:50Z","file_size":1361313,"date_updated":"2018-12-12T10:16:50Z","creator":"system"}],"language":[{"iso":"eng"}],"publication_identifier":{"issn":["14752859"]},"publication_status":"published","volume":16,"issue":"1","pmid":1,"oa_version":"Published Version","abstract":[{"text":"Background: Metabolic engineering and synthetic biology of cyanobacteria offer a promising sustainable alternative approach for fossil-based ethylene production, by using sunlight via oxygenic photosynthesis, to convert carbon dioxide directly into ethylene. Towards this, both well-studied cyanobacteria, i.e., Synechocystis sp PCC 6803 and Synechococcus elongatus PCC 7942, have been engineered to produce ethylene by introducing the ethylene-forming enzyme (Efe) from Pseudomonas syringae pv. phaseolicola PK2 (the Kudzu strain), which catalyzes the conversion of the ubiquitous tricarboxylic acid cycle intermediate 2-oxoglutarate into ethylene. Results: This study focuses on Synechocystis sp PCC 6803 and shows stable ethylene production through the integration of a codon-optimized version of the efe gene under control of the Ptrc promoter and the core Shine-Dalgarno sequence (5\\'-AGGAGG-3\\') as the ribosome-binding site (RBS), at the slr0168 neutral site. We have increased ethylene production twofold by RBS screening and further investigated improving ethylene production from a single gene copy of efe, using multiple tandem promoters and by putting our best construct on an RSF1010-based broad-host-self-replicating plasmid, which has a higher copy number than the genome. Moreover, to raise the intracellular amounts of the key Efe substrate, 2-oxoglutarate, from which ethylene is formed, we constructed a glycogen-synthesis knockout mutant (glgC) and introduced the ethylene biosynthetic pathway in it. Under nitrogen limiting conditions, the glycogen knockout strain has increased intracellular 2-oxoglutarate levels; however, surprisingly, ethylene production was lower in this strain than in the wild-type background. Conclusion: Making use of different RBS sequences, production of ethylene ranging over a 20-fold difference has been achieved. However, a further increase of production through multiple tandem promoters and a broad-host plasmid was not achieved speculating that the transcription strength and the gene copy number are not the limiting factors in our system.","lang":"eng"}],"month":"02","intvolume":" 16","scopus_import":"1","extern":"1","ddc":["579"],"date_updated":"2023-09-20T12:09:21Z","file_date_updated":"2018-12-12T10:16:50Z","_id":"1061","status":"public","pubrep_id":"792","type":"journal_article","tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","short":"CC BY (4.0)"}},{"department":[{"_id":"ToBo"}],"date_updated":"2021-01-12T06:49:10Z","type":"journal_article","status":"public","_id":"1218","issue":"14","volume":82,"publication_status":"published","language":[{"iso":"eng"}],"scopus_import":1,"main_file_link":[{"url":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4959195/","open_access":"1"}],"month":"07","intvolume":" 82","abstract":[{"lang":"eng","text":"Investigating the physiology of cyanobacteria cultured under a diel light regime is relevant for a better understanding of the resulting growth characteristics and for specific biotechnological applications that are foreseen for these photosynthetic organisms. Here, we present the results of a multiomics study of the model cyanobacterium Synechocystis sp. strain PCC 6803, cultured in a lab-scale photobioreactor in physiological conditions relevant for large-scale culturing. The culture was sparged withN2 andCO2, leading to an anoxic environment during the dark period. Growth followed the availability of light. Metabolite analysis performed with 1Hnuclear magnetic resonance analysis showed that amino acids involved in nitrogen and sulfur assimilation showed elevated levels in the light. Most protein levels, analyzed through mass spectrometry, remained rather stable. However, several high-light-response proteins and stress-response proteins showed distinct changes at the onset of the light period. Microarray-based transcript analysis found common patterns of~56% of the transcriptome following the diel regime. These oscillating transcripts could be grouped coarsely into genes that were upregulated and downregulated in the dark period. The accumulated glycogen was degraded in the anaerobic environment in the dark. A small part was degraded gradually, reflecting basic maintenance requirements of the cells in darkness. Surprisingly, the largest part was degraded rapidly in a short time span at the end of the dark period. This degradation could allow rapid formation of metabolic intermediates at the end of the dark period, preparing the cells for the resumption of growth at the start of the light period."}],"oa_version":"Submitted Version","author":[{"orcid":"0000-0001-8619-2223","full_name":"Angermayr, Andreas","last_name":"Angermayr","id":"4677C796-F248-11E8-B48F-1D18A9856A87","first_name":"Andreas"},{"first_name":"Pascal","full_name":"Van Alphen, Pascal","last_name":"Van Alphen"},{"first_name":"Dicle","last_name":"Hasdemir","full_name":"Hasdemir, Dicle"},{"first_name":"Gertjan","last_name":"Kramer","full_name":"Kramer, Gertjan"},{"last_name":"Iqbal","full_name":"Iqbal, Muzamal","first_name":"Muzamal"},{"last_name":"Van Grondelle","full_name":"Van Grondelle, Wilmar","first_name":"Wilmar"},{"first_name":"Huub","last_name":"Hoefsloot","full_name":"Hoefsloot, Huub"},{"first_name":"Younghae","full_name":"Choi, Younghae","last_name":"Choi"},{"full_name":"Hellingwerf, Klaas","last_name":"Hellingwerf","first_name":"Klaas"}],"publist_id":"6117","title":"Culturing synechocystis sp. Strain pcc 6803 with N2 and CO2 in a diel regime reveals multiphase glycogen dynamics with low maintenance costs","citation":{"mla":"Angermayr, Andreas, et al. “Culturing Synechocystis Sp. Strain Pcc 6803 with N2 and CO2 in a Diel Regime Reveals Multiphase Glycogen Dynamics with Low Maintenance Costs.” Applied and Environmental Microbiology, vol. 82, no. 14, American Society for Microbiology, 2016, pp. 4180–89, doi:10.1128/AEM.00256-16.","short":"A. Angermayr, P. Van Alphen, D. Hasdemir, G. Kramer, M. Iqbal, W. Van Grondelle, H. Hoefsloot, Y. Choi, K. Hellingwerf, Applied and Environmental Microbiology 82 (2016) 4180–4189.","ieee":"A. Angermayr et al., “Culturing synechocystis sp. Strain pcc 6803 with N2 and CO2 in a diel regime reveals multiphase glycogen dynamics with low maintenance costs,” Applied and Environmental Microbiology, vol. 82, no. 14. American Society for Microbiology, pp. 4180–4189, 2016.","ama":"Angermayr A, Van Alphen P, Hasdemir D, et al. Culturing synechocystis sp. Strain pcc 6803 with N2 and CO2 in a diel regime reveals multiphase glycogen dynamics with low maintenance costs. Applied and Environmental Microbiology. 2016;82(14):4180-4189. doi:10.1128/AEM.00256-16","apa":"Angermayr, A., Van Alphen, P., Hasdemir, D., Kramer, G., Iqbal, M., Van Grondelle, W., … Hellingwerf, K. (2016). Culturing synechocystis sp. Strain pcc 6803 with N2 and CO2 in a diel regime reveals multiphase glycogen dynamics with low maintenance costs. Applied and Environmental Microbiology. American Society for Microbiology. https://doi.org/10.1128/AEM.00256-16","chicago":"Angermayr, Andreas, Pascal Van Alphen, Dicle Hasdemir, Gertjan Kramer, Muzamal Iqbal, Wilmar Van Grondelle, Huub Hoefsloot, Younghae Choi, and Klaas Hellingwerf. “Culturing Synechocystis Sp. Strain Pcc 6803 with N2 and CO2 in a Diel Regime Reveals Multiphase Glycogen Dynamics with Low Maintenance Costs.” Applied and Environmental Microbiology. American Society for Microbiology, 2016. https://doi.org/10.1128/AEM.00256-16.","ista":"Angermayr A, Van Alphen P, Hasdemir D, Kramer G, Iqbal M, Van Grondelle W, Hoefsloot H, Choi Y, Hellingwerf K. 2016. Culturing synechocystis sp. Strain pcc 6803 with N2 and CO2 in a diel regime reveals multiphase glycogen dynamics with low maintenance costs. Applied and Environmental Microbiology. 82(14), 4180–4189."},"user_id":"3E5EF7F0-F248-11E8-B48F-1D18A9856A87","page":"4180 - 4189","date_published":"2016-07-01T00:00:00Z","doi":"10.1128/AEM.00256-16","date_created":"2018-12-11T11:50:46Z","year":"2016","day":"01","publication":"Applied and Environmental Microbiology","quality_controlled":"1","publisher":"American Society for Microbiology","oa":1,"acknowledgement":"Dutch Ministry of Economic Affairs, Agriculture, and Innovation through the program BioSolar CellsS. Andreas Angermayr,Pascal van Alphen, Klaas J. Hellingwerf\r\nWe thank Naira Quintana (presently at Rousselot, Belgium) for the ini-\r\ntiative at the 10th Cyanobacterial Molecular Biology Workshop\r\n(CMBW), June 2010, Lake Arrowhead, Los Angeles, CA, USA, to start the\r\ncollaborative endeavor reported here. We thank Timo Maarleveld from\r\nCWI/VU (Amsterdam) for a custom-made Python script handling the output from the NMR analysis and for evaluating and visualizing the\r\nseparate metabolites for their evaluation. We thank Rob Verpoorte from\r\nLeiden University (metabolome analysis) and Hans Aerts from the AMC\r\n(proteome analysis) for lab space and equipment. We thank Robert Leh-\r\nmann (Humboldt University Berlin) and Ilka Axmann (University of\r\nDüsseldorf) for sharing the R-code for the LOS transformation of the\r\ntranscript data. We thank Hans C. P. Matthijs from IBED for inspiring\r\ndialogues and insightful thoughts on continuous culturing of cyanobac-\r\nteria. We thank Sandra Waaijenborg for performing the transcript nor-\r\nmalization and Johan Westerhuis from BDA, Jeroen van der Steen and\r\nFilipe Branco dos Santos from MMP, and Lucas Stal from IBED/NIOZ for\r\nhelpful discussions. We thank Milou Schuurmans from MMP for help\r\nwith sampling and glycogen determination. We thank the members of the\r\nRNA Biology & Applied Bioinformatics group at SILS, in particular Selina\r\nvan Leeuwen, Elisa Hoekstra, and Martijs Jonker, for the microarray anal-\r\nysis. We thank the reviewers of this work for their insightful comments\r\nwhich improved the quality of the manuscript. This work, including the efforts of S. Andreas Angermayr, Pascal van\r\nAlphen, and Klaas J. Hellingwerf, was funded by Dutch Ministry of Eco-\r\nnomic Affairs, Agriculture, and Innovation through the program BioSolar\r\nCells."},{"date_created":"2018-12-11T11:52:52Z","issue":"6","doi":"10.1016/j.tibtech.2015.03.009","volume":33,"date_published":"2015-06-01T00:00:00Z","page":"352 - 361","language":[{"iso":"eng"}],"publication":"Trends in Biotechnology","day":"01","publication_status":"published","year":"2015","intvolume":" 33","month":"06","publisher":"Elsevier","scopus_import":1,"quality_controlled":"1","oa_version":"None","abstract":[{"text":"Through metabolic engineering cyanobacteria can be employed in biotechnology. Combining the capacity for oxygenic photosynthesis and carbon fixation with an engineered metabolic pathway allows carbon-based product formation from CO2, light, and water directly. Such cyanobacterial 'cell factories' are constructed to produce biofuels, bioplastics, and commodity chemicals. Efforts of metabolic engineers and synthetic biologists allow the modification of the intermediary metabolism at various branching points, expanding the product range. The new biosynthesis routes 'tap' the metabolism ever more efficiently, particularly through the engineering of driving forces and utilization of cofactors generated during the light reactions of photosynthesis, resulting in higher product titers. High rates of carbon rechanneling ultimately allow an almost-complete allocation of fixed carbon to product above biomass.","lang":"eng"}],"department":[{"_id":"ToBo"}],"title":"Metabolic engineering of cyanobacteria for the synthesis of commodity products","author":[{"id":"4677C796-F248-11E8-B48F-1D18A9856A87","first_name":"Andreas","orcid":"0000-0001-8619-2223","full_name":"Angermayr, Andreas","last_name":"Angermayr"},{"full_name":"Gorchs, Aleix","last_name":"Gorchs","first_name":"Aleix"},{"first_name":"Klaas","last_name":"Hellingwerf","full_name":"Hellingwerf, Klaas"}],"publist_id":"5585","user_id":"3E5EF7F0-F248-11E8-B48F-1D18A9856A87","citation":{"chicago":"Angermayr, Andreas, Aleix Gorchs, and Klaas Hellingwerf. “Metabolic Engineering of Cyanobacteria for the Synthesis of Commodity Products.” Trends in Biotechnology. Elsevier, 2015. https://doi.org/10.1016/j.tibtech.2015.03.009.","ista":"Angermayr A, Gorchs A, Hellingwerf K. 2015. Metabolic engineering of cyanobacteria for the synthesis of commodity products. Trends in Biotechnology. 33(6), 352–361.","mla":"Angermayr, Andreas, et al. “Metabolic Engineering of Cyanobacteria for the Synthesis of Commodity Products.” Trends in Biotechnology, vol. 33, no. 6, Elsevier, 2015, pp. 352–61, doi:10.1016/j.tibtech.2015.03.009.","ama":"Angermayr A, Gorchs A, Hellingwerf K. Metabolic engineering of cyanobacteria for the synthesis of commodity products. Trends in Biotechnology. 2015;33(6):352-361. doi:10.1016/j.tibtech.2015.03.009","apa":"Angermayr, A., Gorchs, A., & Hellingwerf, K. (2015). Metabolic engineering of cyanobacteria for the synthesis of commodity products. Trends in Biotechnology. Elsevier. https://doi.org/10.1016/j.tibtech.2015.03.009","short":"A. Angermayr, A. Gorchs, K. Hellingwerf, Trends in Biotechnology 33 (2015) 352–361.","ieee":"A. Angermayr, A. Gorchs, and K. Hellingwerf, “Metabolic engineering of cyanobacteria for the synthesis of commodity products,” Trends in Biotechnology, vol. 33, no. 6. Elsevier, pp. 352–361, 2015."},"date_updated":"2021-01-12T06:51:46Z","status":"public","type":"journal_article","_id":"1586"},{"citation":{"mla":"Hammar, Petter, et al. “Single-Cell Screening of Photosynthetic Growth and Lactate Production by Cyanobacteria.” Biotechnology for Biofuels, vol. 8, no. 1, 193, BioMed Central, 2015, doi:10.1186/s13068-015-0380-2.","ieee":"P. Hammar et al., “Single-cell screening of photosynthetic growth and lactate production by cyanobacteria,” Biotechnology for Biofuels, vol. 8, no. 1. BioMed Central, 2015.","short":"P. Hammar, A. Angermayr, S. Sjostrom, J. Van Der Meer, K. Hellingwerf, E. Hudson, H. Joensson, Biotechnology for Biofuels 8 (2015).","apa":"Hammar, P., Angermayr, A., Sjostrom, S., Van Der Meer, J., Hellingwerf, K., Hudson, E., & Joensson, H. (2015). Single-cell screening of photosynthetic growth and lactate production by cyanobacteria. Biotechnology for Biofuels. BioMed Central. https://doi.org/10.1186/s13068-015-0380-2","ama":"Hammar P, Angermayr A, Sjostrom S, et al. Single-cell screening of photosynthetic growth and lactate production by cyanobacteria. Biotechnology for Biofuels. 2015;8(1). doi:10.1186/s13068-015-0380-2","chicago":"Hammar, Petter, Andreas Angermayr, Staffan Sjostrom, Josefin Van Der Meer, Klaas Hellingwerf, Elton Hudson, and Hakaan Joensson. “Single-Cell Screening of Photosynthetic Growth and Lactate Production by Cyanobacteria.” Biotechnology for Biofuels. BioMed Central, 2015. https://doi.org/10.1186/s13068-015-0380-2.","ista":"Hammar P, Angermayr A, Sjostrom S, Van Der Meer J, Hellingwerf K, Hudson E, Joensson H. 2015. Single-cell screening of photosynthetic growth and lactate production by cyanobacteria. Biotechnology for Biofuels. 8(1), 193."},"user_id":"2DF688A6-F248-11E8-B48F-1D18A9856A87","author":[{"full_name":"Hammar, Petter","last_name":"Hammar","first_name":"Petter"},{"first_name":"Andreas","id":"4677C796-F248-11E8-B48F-1D18A9856A87","last_name":"Angermayr","full_name":"Angermayr, Andreas","orcid":"0000-0001-8619-2223"},{"first_name":"Staffan","last_name":"Sjostrom","full_name":"Sjostrom, Staffan"},{"first_name":"Josefin","full_name":"Van Der Meer, Josefin","last_name":"Van Der Meer"},{"last_name":"Hellingwerf","full_name":"Hellingwerf, Klaas","first_name":"Klaas"},{"last_name":"Hudson","full_name":"Hudson, Elton","first_name":"Elton"},{"first_name":"Hakaan","last_name":"Joensson","full_name":"Joensson, Hakaan"}],"publist_id":"5537","title":"Single-cell screening of photosynthetic growth and lactate production by cyanobacteria","article_number":"193","year":"2015","has_accepted_license":"1","publication":"Biotechnology for Biofuels","day":"25","date_created":"2018-12-11T11:53:05Z","doi":"10.1186/s13068-015-0380-2","date_published":"2015-11-25T00:00:00Z","oa":1,"quality_controlled":"1","publisher":"BioMed Central","date_updated":"2021-01-12T06:52:04Z","ddc":["570"],"file_date_updated":"2020-07-14T12:45:07Z","department":[{"_id":"ToBo"}],"_id":"1623","tmp":{"legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","image":"/images/cc_by.png","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","short":"CC BY (4.0)"},"type":"journal_article","pubrep_id":"467","status":"public","publication_status":"published","language":[{"iso":"eng"}],"file":[{"access_level":"open_access","relation":"main_file","content_type":"application/pdf","checksum":"172b0b6f4eb2e5c22b7cec1d57dc0107","file_id":"4796","creator":"system","date_updated":"2020-07-14T12:45:07Z","file_size":2914089,"date_created":"2018-12-12T10:10:11Z","file_name":"IST-2016-467-v1+1_s13068-015-0380-2.pdf"}],"volume":8,"issue":"1","abstract":[{"text":"Background\r\nPhotosynthetic cyanobacteria are attractive for a range of biotechnological applications including biofuel production. However, due to slow growth, screening of mutant libraries using microtiter plates is not feasible.\r\nResults\r\nWe present a method for high-throughput, single-cell analysis and sorting of genetically engineered l-lactate-producing strains of Synechocystis sp. PCC6803. A microfluidic device is used to encapsulate single cells in picoliter droplets, assay the droplets for l-lactate production, and sort strains with high productivity. We demonstrate the separation of low- and high-producing reference strains, as well as enrichment of a more productive l-lactate-synthesizing population after UV-induced mutagenesis. The droplet platform also revealed population heterogeneity in photosynthetic growth and lactate production, as well as the presence of metabolically stalled cells.\r\nConclusions\r\nThe workflow will facilitate metabolic engineering and directed evolution studies and will be useful in studies of cyanobacteria biochemistry and physiology.\r\n","lang":"eng"}],"oa_version":"Published Version","scopus_import":1,"intvolume":" 8","month":"11"}]