Phototrophic production of heterologous diterpenoids and a hydroxy-functionalized derivative from Chlamydomonas reinhardtii

Lauersen, Kyle J.; Wichmann, Julian; Baier, Thomas; Kampranis, Sotirios C.; Pateraki, Irini; Møller, Birger Lindberg; Kruse, Olaf

doi:10.1016/j.ymben.2018.07.005

Cited by 104 publications

(135 citation statements)

References 66 publications

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“…Recently, we demonstrated that transgenes could be optimized for expression from the algal nuclear genome by codon optimization and systematic incorporation of the first intron of the C. reinhardtii RuBisCO small subunit II gene to mimic host regulatory structures (Baier et al, 2018b). We have recently used this strategy for overexpression of numerous foreign genes (Lauersen et al, 2016;Lauersen et al, 2018;Wichmann et al, 2018) , as well as re-coding and overexpression of the endogenous photodecarboxylase (CrFAP) (Yunus et al, 2018) for various metabolic engineering activities in this host. Here, the amino acid sequence of the CrBKT was used to generate an optimized synthetic algal transgene, employing this same optimization strategy ( Figure 2b).…”

Section: Intragenic Expression Of β-Ketolase In C Reinhardtiimentioning

confidence: 99%

Turning a green alga red: engineering astaxanthin biosynthesis by intragenic pseudogene revival in Chlamydomonas reinhardtii

Perozeni

Cazzaniga

Baier

et al. 2019

Preprint

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The green alga Chlamydomonas reinhardtii does not synthesize high-value ketocarotenoids like canthaxanthin and astaxanthin, however, a β-carotene ketolase (CrBKT) can be found in its genome.CrBKT is poorly expressed, contains a long C-terminal extension not found in homologues and likely represents a pseudogene in this alga. Here, we used synthetic re-design of this gene to enable its constitutive overexpression from the nuclear genome of C. reinhardtii. Overexpression of the optimized CrBKT extended native carotenoid biosynthesis to generate ketocarotenoids in the algal host causing noticeable changes the green algal colour to a reddish-brown. We found that up to 50% of native carotenoids could be converted into astaxanthin and more than 70% into other ketocarotenoids by robust CrBKT overexpression. Modification of the carotenoid metabolism did not impair growth or biomass productivity of C. reinhardtii, even at high light intensities. Under different growth conditions, the best performing CrBKT overexpression strain was found to reach ketocarotenoid productivities up to 4.5 mg L -1 day -1 . Astaxanthin productivity in engineered C. reinhardtii shown here is competitive with that reported for Haematococcus lacustris (formerly pluvialis) which is currently the main organism cultivated for industrial astaxanthin production. In addition, the extractability and bio-accessibility of these pigments was much higher in cell wall deficient C. reinhardtii than the resting cysts of H. lacustris. Engineered C. reinhardtii strains could thus be a promising alternative to natural astaxanthin producing algal strains and may open the possibility of other tailor-made pigments from this host.

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Section: Intragenic Expression Of β-Ketolase In C Reinhardtiimentioning

confidence: 99%

Turning a green alga red: engineering astaxanthin biosynthesis by intragenic pseudogene revival in Chlamydomonas reinhardtii

Perozeni

Cazzaniga

Baier

et al. 2019

Preprint

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“…Finally, several non-native diterpenoids were successfully produced in C. reinhardtii, by the heterologous expression of diterpene synthases targeted to the algal chloroplast, to convert native GGPP into the diterpenoid products casbene, taxadiene and 13R(+) manoyl oxide [169].…”

Section: Endogenous Up-regulation and Heterologous Expression Of Isopmentioning

confidence: 99%

Potential and Challenges of Improving Photosynthesis in Algae

Vecchi

Barera

Bassi

et al. 2020

Plants

108

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Sunlight energy largely exceeds the energy required by anthropic activities, and therefore its exploitation represents a major target in the field of renewable energies. The interest in the mass cultivation of green microalgae has grown in the last decades, as algal biomass could be employed to cover a significant portion of global energy demand. Advantages of microalgal vs. plant biomass production include higher light-use efficiency, efficient carbon capture and the valorization of marginal lands and wastewaters. Realization of this potential requires a decrease of the current production costs, which can be obtained by increasing the productivity of the most common industrial strains, by the identification of factors limiting biomass yield, and by removing bottlenecks, namely through domestication strategies aimed to fill the gap between the theoretical and real productivity of algal cultures. In particular, the light-to-biomass conversion efficiency represents one of the major constraints for achieving a significant improvement of algal cell lines. This review outlines the molecular events of photosynthesis, which regulate the conversion of light into biomass, and discusses how these can be targeted to enhance productivity through mutagenesis, strain selection or genetic engineering. This review highlights the most recent results in the manipulation of the fundamental mechanisms of algal photosynthesis, which revealed that a significant yield enhancement is feasible. Moreover, metabolic engineering of microalgae, focused upon the development of renewable fuel biorefineries, has also drawn attention and resulted in efforts for enhancing productivity of oil or isoprenoids.

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“…Moreover, isoprenoids metabolic pathways are modulatory, making them easy targets to manipulate in multiple hosts. As an example, reliable non-native di-/tri-/sesquiterpenoids production has been recently achieved in P. tricornutum and C. reinhardtii (Lauersen et al 2016;Wichmann et al 2018;D'Adamo et al 2018;Lauersen et al 2018), increasing their commercialisation potential.…”

Section: Improving the Content Of Bio-active Compoundsmentioning

confidence: 99%

Potential of Microalgae Biomass for the Sustainable Production of Bio-commodities

Perin

Morosinotto

2019

Progress in Botany

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Human activities are causing major negative environmental impacts and the development of sustainable processes for production of commodities is a major urgency. Plants biomass represents a valuable alternative to produce energy and materials but exploiting present crops for commodities production would however require massive resources (i.e. land, water and nutrients), raising serious sustainability concerns. In addition to efforts to improve plants land and resources use efficiency, it is thus fundamental to look for alternative sources of biomass to complement crops. Microalgae are unicellular photosynthetic organisms that show a huge, yet untapped, potential in this context. Microalgae metabolism is powered by photosynthesis and thus uses sunlight, a renewable energy source and the exploitation of microalgae-based products has the potential to provide a beneficial environmental impact. These microorganisms have the ability to synthesise a wide spectrum of bioactive compounds, with many different potential applications (e.g. nutraceutics/pharmaceutics and biofuels). Several, still unresolved, challenges are however present such as the lack of cost-effective cultivation platforms and biomass-harvesting technologies. Moreover, the natural metabolic plasticity of microalgae is not optimized for a production at scale and low biomass productivity and product yields affect competitiveness. Tuning microalgae metabolism to maximize productivity thus represents an unavoidable challenge to reach the theoretical potential of such organisms.

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Phototrophic production of heterologous diterpenoids and a hydroxy-functionalized derivative from Chlamydomonas reinhardtii

Cited by 104 publications

References 66 publications

Turning a green alga red: engineering astaxanthin biosynthesis by intragenic pseudogene revival in Chlamydomonas reinhardtii

Turning a green alga red: engineering astaxanthin biosynthesis by intragenic pseudogene revival in Chlamydomonas reinhardtii

Potential and Challenges of Improving Photosynthesis in Algae

Potential of Microalgae Biomass for the Sustainable Production of Bio-commodities

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