Biomass generation and heterologous isoprenoid milking from engineered microalgae grown in anaerobic membrane bioreactor effluent

Freitas, Bárbara Bastos de; Overmans, Sebastian; Medina, Julie Sanchez; Hong, Pei‐Ying; Lauersen, Kyle J.

doi:10.1016/j.watres.2022.119486

Cited by 18 publications

(20 citation statements)

References 42 publications

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“…Preculturing prior to growth analysis was performed by inoculation of cells to shake flasks with MA2 medium for 4 days, then cells were resuspended according to desired densities in 400 mL flasks prior to growth analysis in Algem photobioreactors (Algenuity©, UK). Cultures in biological duplicates with starting cell concentrations of 3×10 6 cells mL −1 were cultivated in environmental simulations of the mid Red Sea coast conditions of Thuwal, Saudi Arabia (22.3046N, 39.1022E) as recently reported (de Freitas et al, 2023) and compared with control cultures using 12:12 h light:dark or 24 h constant 1500 μ E illumination at 42 °C. All cultures received continuous 7% CO 2 in air at a 25 mL min −1 flow rate.…”

Section: Methodsmentioning

confidence: 99%

Cultivation of the polyextremophileCyanidioschyzon merolae10D during summer conditions on the coast of the Red Sea and its adaptation to hypersaline sea water

Villegas-Valencia

González-Portela

Freitas

et al. 2023

Preprint

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The west coast of Saudi Arabia borders the Red Sea, which maintains high average temperatures and increased salinity compared to other seas or oceans. Summer conditions in the Arabian Peninsula may exceed the temperature tolerance of most currently cultivated microalgae. The Cyanidiales are polyextremophilic red algae whose native habitats are at the edges of acidic hot springs. Cyanidioschyzon merolae 10D has recently emerged as an interesting model organism capable of high-cell density cultivation on pure CO2 with optimal growth at 42 degrees C and low pH between 0.5-2. C. merolae biomass has an interesting macromolecular composition, is protein rich, and contains valuable bio-products like heat-stable phycocyanin, carotenoids, beta-glucan, and starch. Here, photobioreactors were used to model C. merolae 10D growth performance in simulated environmental conditions of the mid-Red Sea coast across four seasons, it was then grown at various scales outdoors in Thuwal, Saudi Arabia during the Summer of 2022. We show that C. merolae 10D is amenable to cultivation with industrial-grade nutrient and CO2 inputs outdoors in this location and that its biomass is relatively constant in biochemical composition across culture conditions. We also show the adaptation of C. merolae 10D to high salinity levels of those found in Red Sea waters and conducted further modeled cultivations in nutrient enriched local sea water. It was determined that salt-water adapted C. merolae 10D could be cultivated with reduced nutrient inputs in local conditions. The results presented here indicate this may be a promising alternative species for algal bioprocesses in outdoor conditions in extreme desert summer environments.

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Section: Methodsmentioning

confidence: 99%

Cultivation of the polyextremophileCyanidioschyzon merolae10D during summer conditions on the coast of the Red Sea and its adaptation to hypersaline sea water

Villegas-Valencia

González-Portela

Freitas

et al. 2023

Preprint

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“…For the green alga, catalytic activity of the IbIspS mediated by temperature appears to be the most significant factor in isoprene production, and that optimal enzyme activity is likely outside its range of growth temperatures. However, daily high-temperature peaks are tolerated for C. reinhardtii, as recently shown in environmental modelling in the extreme environment of the Red Sea coast (de Freitas et al, 2023). It could, therefore, be possible to create cultivation parameters that balance cell growth densities and temperature in lower and consistent light conditions to maximize isoprene yields.…”

Section: Temperature Affects Engineered Algal Isoprene Production Mor...mentioning

confidence: 98%

“…As alternatives to fermentative hosts, algae can produce heterologous products of interest directly from CO 2 as a carbon source, while consuming nitrogen and phosphorous sources in water (de Freitas et al, 2023). These properties make microalgae interesting for sustainable bio-production concepts that convert waste to value.…”

Section: Chlamydomonas As a Host For Heterologous Isoprenoid Productionmentioning

confidence: 99%

“…It was recently shown that heterologous patchoulol production was lower when cells were exposed to modeled higher temperature and light environmental conditions (de Freitas et al, 2023), suggesting that both temperature and light regimes impact isoprenoid biosynthesis in the alga. Isoprene synthases have optimal catalytic activities at ~42 • C (Li et al, 2019).…”

Section: Temperature Affects Engineered Algal Isoprene Production Mor...mentioning

confidence: 99%

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Engineered production of isoprene from the model green microalga Chlamydomonas reinhardtii

Yahya

Wellman

Overmans

et al. 2023

Metabolic Engineering Communications

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“…As alternatives to fermentative hosts, algal chassis can produce heterologous products of interest directly from CO2 as a carbon source, while consuming nitrogen and phosphorous sources in water (de Freitas et al, 2023). These properties make microalgae interesting for sustainable bioproduction concepts that convert waste to value.…”

Section: Chlamydomonas As a Host For Heterologous Isoprenoid Productionmentioning

confidence: 99%

Engineered production of isoprene from the model green microalgaChlamydomonas reinhardtii

Yahya

Wellman

Overmans

et al. 2023

Preprint

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Isoprene is a clear, colorless, volatile 5-carbon hydrocarbon that is one monomer of all cellular isoprenoids and a platform chemical with multiple applications in industry. Many plants have evolved isoprene synthases (IspSs) with the capacity to liberate isoprene from dimethylallyl pyrophosphate (DMAPP) as part of cellular protection mechanisms. Isoprene is hydrophobic and volatile, rapidly leaves plant tissues and is one of the main carbon emission sources from vegetation globally. The universality of isoprenoid metabolism allows volatile isoprene production from microbes expressing heterologous IspSs. Here, we compared heterologous overexpression from the nuclear genome and localization into the plastid of four plant terpene synthases (TPs) in the green microalga Chlamydomonas reinhardtii. Using sealed vial mixotrophic cultivation, direct quantification of isoprene production was achieved from the headspace of living cultures, with the highest isoprene production observed in algae expressing the Ipomoea batatas IspS. Perturbations of the downstream carotenoid pathway through keto carotenoid biosynthesis enhanced isoprene titers, which could be further enhanced by increasing flux towards DMAPP through heterologous co-expression of a yeast isopentenyl-PP delta isomerase. Multiplexed controlled-environment testing revealed that cultivation temperature, rather than illumination intensity, was the main factor affecting isoprene yield from the engineered alga. This is the first report of heterologous isoprene production from a eukaryotic alga and sets a foundation for further exploration of carbon conversion to this commodity chemical.

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Biomass generation and heterologous isoprenoid milking from engineered microalgae grown in anaerobic membrane bioreactor effluent

Cited by 18 publications

References 42 publications

Cultivation of the polyextremophileCyanidioschyzon merolae10D during summer conditions on the coast of the Red Sea and its adaptation to hypersaline sea water

Cultivation of the polyextremophileCyanidioschyzon merolae10D during summer conditions on the coast of the Red Sea and its adaptation to hypersaline sea water

Engineered production of isoprene from the model green microalga Chlamydomonas reinhardtii

Engineered production of isoprene from the model green microalgaChlamydomonas reinhardtii

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