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