Eukaryotic microalgae as hosts for light-driven heterologous isoprenoid production
Eukaryotic microalgae hold incredible metabolic potential for the sustainable production of heterologous isoprenoid products. Recent advances in algal engineering have enabled the demonstration of prominent examples of heterologous isoprenoid production.
Isoprenoids, also known as terpenes or terpenoids, are the largest class of natural chemicals, with a vast diversity of structures and biological roles. Some have high-value in human-use applications, although may be found in their native contexts in low abundance or be difficult to extract and purify. Heterologous production of isoprenoid compounds in heterotrophic microbial hosts such as bacteria or yeasts has been an active area of research for some time and is now a mature technology. Eukaryotic microalgae represent sustainable alternatives to these hosts for biotechnological production processes as their cultivation can be driven by light and freely available CO2 as a carbon source. Their photosynthetic lifestyles require metabolic architectures structured towards the generation of associated isoprenoids (carotenoids, phytol) which participate in photon capture, energy dissipation, and electron transfer. Eukaryotic microalgae should, therefore, contain inherently high capacities for the generation of heterologous isoprenoid products. Although engineering strategies in eukaryotic microalgae have lagged behind the more genetically tractable bacteria and yeasts, recent advances in algal engineering concepts have demonstrated prominent examples of light-driven heterologous isoprenoid production from these photosynthetic hosts. This work seeks to provide practical insights into the choice of eukaryotic microalgae as biotechnological chassis. Recent reports of advances in algal engineering for heterologous isoprenoid production are highlighted as encouraging examples that promote their expanded use as sustainable green-cell factories. Current state of the art, limitations, and future challenges are also discussed.
KeywordsMicroalgae Chlamydomonas reinhardtii Phaeodactylum tricornutum Terpenoids Isoprenoids Cytochrome P450s
Geranyl pyrophosphate (synthase)
Farnesyl pyrophosphate (synthase)
Geranylgeranyl pyrophosphate (synthase)
Yellow fluorescent protein
Cyan fluorescent protein
Red fluorescent protein
Cytochrome P450 monooxygenase
This work has been supported by the technology platform and infrastructure at the Center for Biotechnology (CeBiTec) of Bielefeld University. Sincere thanks to Dr. Thomas Baier for critical reading of this manuscript and those mentioned in the text who provided pictures.
Compliance with ethical standards
Conflict of interest
The author declares no conflict of interest.
- Archibald JM (2012) the evolution of algae by secondary and tertiary endosymbiosis. In: Piganeau G (ed) Advances in botanical research. Elsevier, New York, pp 87–118Google Scholar
- Black JB, Perez-Pinera P, Gersbach CA (2017) Mammalian synthetic biology: engineering biological systems. Annu Rev Biomed Eng 19:249–277. https://doi.org/10.1146/annurev-bioeng-071516-044649 CrossRefPubMedGoogle Scholar
- Bogen C, Al-Dilaimi A, Albersmeier A et al (2013) Reconstruction of the lipid metabolism for the microalga Monoraphidium neglectum from its genome sequence reveals characteristics suitable for biofuel production. BMC Genomics 14:926. https://doi.org/10.1186/1471-2164-14-926 CrossRefPubMedPubMedCentralGoogle Scholar
- Bohlmann J, Crock J, Jetter R, Croteau R (1998) Terpenoid-based defenses in conifers: cDNA cloning, characterization, and functional expression of wound-inducible (E)-alpha-bisabolene synthase from grand fir (Abies grandis). Proc Natl Acad Sci USA 95:6756–6761. https://doi.org/10.1073/pnas.95.12.6756 CrossRefPubMedGoogle Scholar
- Buckingham J, Macdonald FM, Bradley HM et al (1994) Dictionary of natural products, 1st edn. Chapman and Hall, LondonGoogle Scholar
- Dong B, Hu HH, Li ZF et al (2017) A novel bicistronic expression system composed of the intraflagellar transport protein gene ift25 and FMDV 2A sequence directs robust nuclear gene expression in Chlamydomonas reinhardtii. Appl Microbiol Biotechnol 101:4227–4245. https://doi.org/10.1007/s00253-017-8177-9 CrossRefPubMedGoogle Scholar
- Gruchattka E, Kayser O (2015) In vivo validation of in silico predicted metabolic engineering strategies in yeast: disruption of α-ketoglutarate dehydrogenase and expression of ATP-citrate lyase for terpenoid production. PLoS One 10:e0144981. https://doi.org/10.1371/journal.pone.0144981 CrossRefPubMedPubMedCentralGoogle Scholar
- Hallmann A (2007) Algal transgenics and biotechnology. Transgenic Plant J 1:81–98Google Scholar
- Kirby J, Keasling JD (2009) Biosynthesis of plant isoprenoids: perspectives for microbial engineering. Annu Rev Plant Biol 60:335–355. https://doi.org/10.1146/annurev.arplant.043008.091955 CrossRefPubMedGoogle Scholar
- Lauersen KJ, Vanderveer TL, Berger H et al (2013b) Ice recrystallization inhibition mediated by a nuclear-expressed and -secreted recombinant ice-binding protein in the microalga Chlamydomonas reinhardtii. Appl Microbiol Biotechnol 97:9763–9772. https://doi.org/10.1007/s00253-013-5226-x CrossRefPubMedGoogle Scholar
- Pateraki I, Heskes AM, Hamberger B (2015) Cytochromes P450 for Terpene Functionalisation and Metabolic Engineering. In: Advances in biochemical engineering/biotechnology. pp 107–139Google Scholar
- Poliner E, Pulman JA, Zienkiewicz K et al (2018b) A toolkit for Nannochloropsis oceanica CCMP1779 enables gene stacking and genetic engineering of the eicosapentaenoic acid pathway for enhanced long-chain polyunsaturated fatty acid production. Plant Biotechnol J 16:298–309. https://doi.org/10.1111/pbi.12772 CrossRefPubMedGoogle Scholar
- Schwender J, Seemann M, Lichtenthaler HK, Rohmer M (1996) Biosynthesis of isoprenoids (carotenoids, sterols, prenyl side-chains of chlorophylls and plastoquinone) via a novel pyruvate/glyceraldehyde 3-phosphate non-mevalonate pathway in the green alga Scenedesmus obliquus. Biochem J 316:73–80. https://doi.org/10.1042/bj3160073 CrossRefPubMedPubMedCentralGoogle Scholar