Metabolic engineering of Synechocystis sp. PCC 6803 for enhanced ethanol production based on flux balance analysis

Abstract

Synechocystis sp. PCC 6803 is an attractive host for bio-ethanol production due to its ability to directly convert atmospheric carbon dioxide into ethanol using photosystems. To enhance ethanol production in Synechocystis sp. PCC 6803, metabolic engineering was performed based on in silico simulations, using the genome-scale metabolic model. Comprehensive reaction knockout simulations by flux balance analysis predicted that the knockout of NAD(P)H dehydrogenase enhanced ethanol production under photoautotrophic conditions, where ammonium is the nitrogen source. This deletion inhibits the re-oxidation of NAD(P)H, which is generated by ferredoxin-NADP+ reductase and imposes re-oxidation in the ethanol synthesis pathway. The effect of deleting the ndhF1 gene, which encodes NADH dehydrogenase subunit 5, on ethanol production was experimentally evaluated using ethanol-producing strains of Synechocystis sp. PCC 6803. The ethanol titer of the ethanol-producing ∆ndhF1 strain increased by 145%, compared with that of the control strain.

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Acknowledgements

We would like to thank Prof. Sonoike (Waseda University) for providing the ΔndhF1 strain of PCC 6803. This work was supported by Grant in Aid for Scientific Research (A) No. 24246134. This work was also supported by JSPS KAKENHI Grant Number 16H06559.

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Correspondence to Hiroshi Shimizu.

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Yoshikawa, K., Toya, Y. & Shimizu, H. Metabolic engineering of Synechocystis sp. PCC 6803 for enhanced ethanol production based on flux balance analysis. Bioprocess Biosyst Eng 40, 791–796 (2017). https://doi.org/10.1007/s00449-017-1744-8

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Keywords

  • Ethanol production
  • Flux balance analysis
  • Genome-scale metabolic model
  • NdhF1 deletion
  • Synechocystis sp. PCC 6803