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Production of 23BD in 7942 was improved by altering the ribosomal binding site. They utilized a combinatorial approach with 4 different RBS sequences. A broad range in 23BD production from 207 mg/L to 496 mg/L was observed. This demonstrates the efficacy of this approach and the importance of regulating gene expression for achieving high titer production.
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5• Oliver, J.W., Machado, I.M., Yoneda, H., Atsumi, S., Combinatorial optimization of cyanobacterial 2,3-butanediol production. Metab Eng 22 (2014), 76–82 Production of 23BD in 7942 was improved by altering the ribosomal binding site. They utilized a combinatorial approach with 4 different RBS sequences. A broad range in 23BD production from 207 mg/L to 496 mg/L was observed. This demonstrates the efficacy of this approach and the importance of regulating gene expression for achieving high titer production.
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1 was investigated and strong control of gene expression was demonstrated. Constitutive promoters in two of the genes in the 23BD pathway (alsS and adh) were discovered and gene order was shown to be critical.
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1 was investigated and strong control of gene expression was demonstrated. Constitutive promoters in two of the genes in the 23BD pathway (alsS and adh) were discovered and gene order was shown to be critical.
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Previous work had identified strategies for sugar utilization in 7942. This work was expanded upon by demonstrating 23BD production from engineered 7942 strains under various light conditions (light, dark, and diurnal) with glucose or xylose. From both the glucose (galP) and xylose (xylEAB) strains, a significant increase in both biomass accumulation, and 23BDO production was observed.
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9•• McEwen, J.T., Kanno, M., Atsumi, S., 2,3 Butanediol production in an obligate photoautotrophic cyanobacterium in dark conditions via diverse sugar consumption. Metab Eng 36 (2016), 28–36 Previous work had identified strategies for sugar utilization in 7942. This work was expanded upon by demonstrating 23BD production from engineered 7942 strains under various light conditions (light, dark, and diurnal) with glucose or xylose. From both the glucose (galP) and xylose (xylEAB) strains, a significant increase in both biomass accumulation, and 23BDO production was observed.
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McEwen, J.T.1
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A biosynthetic pathway of 1,3-propanediol was established in 7942. The addition of a glycerol dehydratase allowed for the conversion of glycerol to 1,3-propanediol. The engineered strain produced 288 mg/L after 14 days under continuous light conditions.
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10•• Hirokawa, Y., Maki, Y., Tatsuke, T., Hanai, T., Cyanobacterial production of 1,3-propanediol directly from carbon dioxide using a synthetic metabolic pathway. Metab Eng 34 (2016), 97–103 A biosynthetic pathway of 1,3-propanediol was established in 7942. The addition of a glycerol dehydratase allowed for the conversion of glycerol to 1,3-propanediol. The engineered strain produced 288 mg/L after 14 days under continuous light conditions.
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Hirokawa, Y.1
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Two genes from E. coli (xylAB) were installed into 6803 to utilize xylose. The addition of xylose consumption genes and xylose improved ethylene production by 64%.
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21• Lee, T-C., Xiong, W., Paddock, T., Carrieri, D., Chang, F., Chiu, H-F., Ungerer, J., Juo, S-H.H., Maness, P-C., Yu, J., Engineered xylose utilization enhances bio-products productivity in the cyanobacterium Synechocystis sp. PCC 6803. Metab Eng 30 (2015), 179–189 Two genes from E. coli (xylAB) were installed into 6803 to utilize xylose. The addition of xylose consumption genes and xylose improved ethylene production by 64%.
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99-99 Lactic acid production in 6803 was improved by optimizing gene expression, increasing carbon flux and eliminating competing pathways. They were further able to increase production by optimizing Ldh through site directed mutagenesis. Carbon partitioning to lactic acid was increased from 5% to over 50%.
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33• Angermayr, S.A., van der Woude, A.D., Correddu, D., Vreugdenhil, A., Verrone, V., Hellingwerf, K.J., Exploring metabolic engineering design principles for the photosynthetic production of lactic acid by Synechocystis sp. PCC6803. Biotechnol Biofuels, 7, 2014 99-99 Lactic acid production in 6803 was improved by optimizing gene expression, increasing carbon flux and eliminating competing pathways. They were further able to increase production by optimizing Ldh through site directed mutagenesis. Carbon partitioning to lactic acid was increased from 5% to over 50%.
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A pathway from C. fritschii PCC 9212 was introduced into 7002 to produce P3HB4HB. 3HB-CoA and 4HB-CoA are condensed into copolymer by polyhydroxyalkanoate synthase. The production was 4.5% of total DCW with 12% 4HB composition.
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