{"title":"Ethylene production with engineered Synechocystis sp PCC 6803 strains","pubrep_id":"792","status":"public","user_id":"c635000d-4b10-11ee-a964-aac5a93f6ac1","doi":"10.1186/s12934-017-0645-5","article_processing_charge":"No","date_created":"2018-12-11T11:49:56Z","date_updated":"2023-09-20T12:09:21Z","license":"https://creativecommons.org/licenses/by/4.0/","file_date_updated":"2018-12-12T10:16:50Z","citation":{"ieee":"V. Veetil, A. Angermayr, and K. Hellingwerf, “Ethylene production with engineered Synechocystis sp PCC 6803 strains,” <i>Microbial Cell Factories</i>, vol. 16, no. 1. BioMed Central, 2017.","chicago":"Veetil, Vinod, Andreas Angermayr, and Klaas Hellingwerf. “Ethylene Production with Engineered Synechocystis Sp PCC 6803 Strains.” <i>Microbial Cell Factories</i>. BioMed Central, 2017. <a href=\"https://doi.org/10.1186/s12934-017-0645-5\">https://doi.org/10.1186/s12934-017-0645-5</a>.","ista":"Veetil V, Angermayr A, Hellingwerf K. 2017. Ethylene production with engineered Synechocystis sp PCC 6803 strains. Microbial Cell Factories. 16(1), 34.","apa":"Veetil, V., Angermayr, A., &#38; Hellingwerf, K. (2017). Ethylene production with engineered Synechocystis sp PCC 6803 strains. <i>Microbial Cell Factories</i>. BioMed Central. <a href=\"https://doi.org/10.1186/s12934-017-0645-5\">https://doi.org/10.1186/s12934-017-0645-5</a>","ama":"Veetil V, Angermayr A, Hellingwerf K. Ethylene production with engineered Synechocystis sp PCC 6803 strains. <i>Microbial Cell Factories</i>. 2017;16(1). doi:<a href=\"https://doi.org/10.1186/s12934-017-0645-5\">10.1186/s12934-017-0645-5</a>","short":"V. Veetil, A. Angermayr, K. Hellingwerf, Microbial Cell Factories 16 (2017).","mla":"Veetil, Vinod, et al. “Ethylene Production with Engineered Synechocystis Sp PCC 6803 Strains.” <i>Microbial Cell Factories</i>, vol. 16, no. 1, 34, BioMed Central, 2017, doi:<a href=\"https://doi.org/10.1186/s12934-017-0645-5\">10.1186/s12934-017-0645-5</a>."},"language":[{"iso":"eng"}],"extern":"1","intvolume":"        16","file":[{"file_id":"5240","creator":"system","date_updated":"2018-12-12T10:16:50Z","file_size":1361313,"file_name":"IST-2017-792-v1+1_s12934-017-0645-5.pdf","content_type":"application/pdf","relation":"main_file","access_level":"open_access","date_created":"2018-12-12T10:16:50Z"}],"publication_identifier":{"issn":["14752859"]},"publist_id":"6325","date_published":"2017-02-23T00:00:00Z","volume":16,"abstract":[{"lang":"eng","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."}],"isi":1,"month":"02","ddc":["579"],"tmp":{"short":"CC BY (4.0)","legal_code_url":"https://creativecommons.org/licenses/by/4.0/legalcode","name":"Creative Commons Attribution 4.0 International Public License (CC-BY 4.0)","image":"/images/cc_by.png"},"issue":"1","oa_version":"Published Version","quality_controlled":"1","external_id":{"isi":["000397733000001"],"pmid":["28231787"]},"type":"journal_article","pmid":1,"publication_status":"published","_id":"1061","publisher":"BioMed Central","has_accepted_license":"1","day":"23","article_number":"34","publication":"Microbial Cell Factories","oa":1,"scopus_import":"1","author":[{"full_name":"Veetil, Vinod","last_name":"Veetil","first_name":"Vinod"},{"last_name":"Angermayr","orcid":"0000-0001-8619-2223","first_name":"Andreas","id":"4677C796-F248-11E8-B48F-1D18A9856A87","full_name":"Angermayr, Andreas"},{"first_name":"Klaas","last_name":"Hellingwerf","full_name":"Hellingwerf, Klaas"}],"year":"2017"}