Biotechnology of cyanobacterial isoprene production
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Heterologous cyanobacterial production of isoprene (C5H8) presents an opportunity to develop renewable resources for fuel and industrial chemicals. Isoprene can be generated photosynthetically in these microorganisms from dimethylallyl-diphosphate (DMAPP) by the recombinant enzyme isoprene synthase (ISPS), as a transgenic product of the isoprenoid biosynthetic pathway. The present work sought to combine recent enhancements in the cellular level of reactant (DMAPP) and enzyme (ISPS), as a means in the further development of this technology. This objective was approached upon the heterologous overexpression of fni, an isopentenyl isomerase from Streptococcus pneumoniae, which increased the amount of the DMAPP reactant at the expense of its isomer, isopentenyl-diphosphate (IPP), in the cells. In addition, the cellular concentration of ISPS was substantially enhanced upon expression of the ISPS gene, as a fusion construct with the highly expressed in cyanobacteria cpcB gene, encoding the abundant β-subunit of phycocyanin. Synergy between these two modifications, i.e., enhancement in DMAPP substrate availability and enhancement in the concentration of the ISPS enzyme, improved the isoprene-to-biomass production ratio in cyanobacteria from 0.2:1 mg g−1 (w:w), attained with the ISPS transgene alone, up to 12.3:1 mg g−1 (w:w), measured when the combined two modifications were applied to the same cell. This is the highest verifiable yield of heterologous photosynthetic isoprene production reported so far. Findings in this work constitute a step forward in the development of the cyanobacterial biotechnology for isoprene production.
KeywordsBioenergy Isoprene Isopentenyl diphosphate isomerase Metabolic engineering Synechocystis PCC 6803 Synthetic biology
The work was conducted in partial satisfaction of the requirements for the degree of Doctor of Philosophy by JEC, who was supported by a graduate student fellowship from the NSF Sage IGERT program. Authors wish to thank Dr. Cinzia Formighieri for making the Fni-SmR construct.
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Conflict of interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
- Branco Dos Santos F, Du W, Hellingwerf KJ (2016) Synechocystis: not just a plug-bug for CO2, but a green E. coli. Front Bioeng Biotechnol 2:36Google Scholar
- Eaton-Rye JJ (2004) The construction of gene knockouts in the cyanobacterium Synechocystis sp. PCC 6803. In Methods of Molecular Biology, Vol 274: Photosynthesis Protocols (Carpentier R, Ed) pp 309–324, Humana Press, Totowa, NJGoogle Scholar
- Glazer AN, Lundell DJ, Yamanaka G, Williams RC (1983) The structure of a “simple” phycobilisome. Ann Microbiol (Paris) 134B(1):159–180Google Scholar
- Knoot CJ, Ungerer J, Wangikar PP, Pakrasi HB (2018) Cyanobacteria: promising biocatalysts for sustainable chemical production. J Biol Chem 293:5044-5052. https://doi.org/10.1074/jbc.R117.815886
- Leonard E, Ajikumar PK, Thayer K, Xiao WH, Mo JD, Tidor B, Stephanopoulos G, Prather KLJ (2010) Combining metabolic and protein engineering of a terpenoid biosynthetic pathway for overproduction and selectivity control. Proc Natl Acad Sci U S A 107(31):13654–13659CrossRefPubMedPubMedCentralGoogle Scholar