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Photosynthetic conversion of CO2 to farnesyl diphosphate-derived phytochemicals (amorpha-4,11-diene and squalene) by engineered cyanobacteria
- Choi, Sun Young, Lee, Hyun Jeong, Choi, Jaeyeon, Kim, Jiye, Sim, Sang Jun, Um, Youngsoon, Kim, Yunje, Lee, Taek Soon, Keasling, Jay D., Woo, Han Min
- Biotechnology for biofuels 2016 v.9 no.1 pp. 202
- Synechococcus sp. PCC 7942, biochemical pathways, carbon dioxide, cell growth, enzymes, genes, growth retardation, hosts, isoprenoids, metabolic engineering, phosphates, photosynthesis, phytopharmaceuticals, squalene, value added
- BACKGROUND: Metabolic engineering of cyanobacteria has enabled photosynthetic conversion of CO₂ to value-added chemicals as bio-solar cell factories. However, the production levels of isoprenoids in engineered cyanobacteria were quite low, compared to other microbial hosts. Therefore, modular optimization of multiple gene expressions for metabolic engineering of cyanobacteria is required for the production of farnesyl diphosphate-derived isoprenoids from CO₂. RESULTS: Here, we engineered Synechococcus elongatus PCC 7942 with modular metabolic pathways consisting of the methylerythritol phosphate pathway enzymes and the amorphadiene synthase for production of amorpha-4,11-diene, resulting in significantly increased levels (23-fold) of amorpha-4,11-diene (19.8 mg/L) in the best strain relative to a parental strain. Replacing amorphadiene synthase with squalene synthase led to the synthesis of a high amount of squalene (4.98 mg/L/OD₇₃₀). Overexpression of farnesyl diphosphate synthase is the most critical factor for the significant production, whereas overexpression of 1-deoxy-D-xylulose 5-phosphate reductase is detrimental to the cell growth and the production. Additionally, the cyanobacterial growth inhibition was alleviated by expressing a terpene synthase in S. elongatus PCC 7942 strain with the optimized MEP pathway only (SeHL33). CONCLUSIONS: This is the first demonstration of photosynthetic production of amorpha-4,11-diene from CO₂ in cyanobacteria and production of squalene in S. elongatus PCC 7942. Our optimized modular OverMEP strain (SeHL33) with either co-expression of ADS or SQS demonstrated the highest production levels of amorpha-4,11-diene and squalene, which could expand the list of farnesyl diphosphate-derived isoprenoids from CO₂ as bio-solar cell factories.