Heterologous expression of phosphite dehydrogenase in the chloroplast or nucleus enables phosphite utilization and genetic selection in Picochlorum spp.

Dahlin, Lukas R.; Guarnieri, Michael T.

doi:10.1016/j.algal.2021.102604

Cited by 10 publications

(12 citation statements)

References 42 publications

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“…reinhardtii , the chloroplast genome of the red alga Cyanidioschyzon merolae 10D has been transformed, 36 , 54 as has the fast-growing green Picochlorum spp. 55 (discussed later). Much remains to be understood regarding transgene expression in algae, and there are numerous species-specific features that require consideration for each organism in which engineering is a goal.…”

Section: Key Developments In Engineering Algal Genomesmentioning

confidence: 94%

The synthetic future of algal genomes

Goold,

Moseley,

Lauersen

2024

Cell Genomics

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Section: Key Developments In Engineering Algal Genomesmentioning

confidence: 94%

The synthetic future of algal genomes

Goold,

Moseley,

Lauersen

2024

Cell Genomics

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“…One strategy for reducing contamination of algal cultures is the introduction of capacity for metabolism of inorganic phosphite as a phosphorous source. This has been shown in numerous organisms, including C. reinhardtii, to reduce contamination and act as a selection agent (López-Arredondo and Herrera-Estrella, 2012;Loera-Quezada et al, 2016;Changko et al, 2020;Cutolo et al, 2020;Dahlin and Guarnieri, 2022). Engineered expression of Pseudomonas stutzeri WM88 phosphite NAD + oxidoreductase ptxD from either the chloroplast or nuclear genomes of algae has been shown to confer the ability to metabolize phosphite (López-Arredondo and Herrera-Estrella, 2012;Changko et al, 2020;Cutolo et al, 2020;Dahlin and Guarnieri, 2022).…”

Section: Introductionmentioning

confidence: 99%

Combinatorial Engineering Enables Photoautotrophic Growth in High Cell Density Phosphite-Buffered Media to Support Engineered Chlamydomonas reinhardtii Bio-Production Concepts

et al. 2022

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Chlamydomonas reinhardtii has emerged as a powerful green cell factory for metabolic engineering of sustainable products created from the photosynthetic lifestyle of this microalga. Advances in nuclear genome modification and transgene expression are allowing robust engineering strategies to be demonstrated in this host. However, commonly used lab strains are not equipped with features to enable their broader implementation in non-sterile conditions and high-cell density concepts. Here, we used combinatorial chloroplast and nuclear genome engineering to augment the metabolism of the C. reinhardtii strain UVM4 with publicly available genetic tools to enable the use of inorganic phosphite and nitrate as sole sources of phosphorous and nitrogen, respectively. We present recipes to create phosphite-buffered media solutions that enable high cell density algal cultivation. We then combined previously reported engineering strategies to produce the heterologous sesquiterpenoid patchoulol to high titers from our engineered green cell factories and show these products are possible to produce in non-sterile conditions. Our work presents a straightforward means to generate C. reinhardtii strains for broader application in bio-processes for the sustainable generation of products from green microalgae.

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“…This has been shown in numerous organisms, including C. reinhardtii, to reduce contamination and act as a selection agent (López-Arredondo and Herrera-Estrella, 2012;Loera-Quezada et al, 2016;Changko et al, 2020;Cutolo et al, 2020;Dahlin and Guarnieri, 2022). Engineered expression of Pseudomonas stutzeri WM88 phosphite NAD+ oxidoreductase ptxD from either the chloroplast or nuclear genomes of algae has been shown to confer the ability to metabolize phosphite (López-Arredondo and Herrera-Estrella, 2012;Changko et al, 2020;Cutolo et al, 2020;Dahlin and Guarnieri, 2022). To date, demonstrated advances in nuclear transgene expression for metabolic engineering described above have not incorporated combinatorial engineering with chloroplast expression constructs in the same strain.…”

Section: Introductionmentioning

confidence: 99%

“…One strategy for reducing contamination of algal cultures is the introduction of metabolic capacity for metabolism of inorganic phosphite as a phosphorous source. This has been shown in numerous organisms, including C. reinhardtii , to reduce contamination and act as a selection agent (López-Arredondo and Herrera-Estrella, 2012; Loera-Quezada et al, 2016; Changko et al, 2020; Cutolo et al, 2020; Dahlin and Guarnieri, 2022). Engineered expression of Pseudomonas stutzeri WM88 phosphite NAD+ oxidoreductase ptxD from either the chloroplast or nuclear genomes of algae has been shown to confer the ability to metabolize phosphite (López-Arredondo and Herrera-Estrella, 2012; Changko et al, 2020; Cutolo et al, 2020; Dahlin and Guarnieri, 2022).…”

Section: Introductionmentioning

confidence: 99%

Combinatorial engineering for photoautotrophic production of recombinant products from the green microalga Chlamydomonas reinhardtii

Abdallah

Wellman

Overmans

et al. 2022

Preprint

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Chlamydomonas reinhardtii has emerged as a powerful green cell factory for metabolic engineering of sustainable products created from the photosynthetic lifestyle of this microalga. Advances in nuclear genome and transgene expression engineering are allowing robust engineering strategies to be demonstrated in this host. However, commonly used lab strains are not equipped with features to enable their broader implementation in non-sterile conditions and high-cell density concepts. Here, we use combinatorial chloroplast and nuclear genome engineering to complement the C. reinhardtii strain UVM4 with publicly available genetic tools to enable the use of inorganic phosphite and nitrate as a sole the source phosphorous and nitrogen, respectively. We present recipes to create phosphite-buffered media solutions that enable high cell density algal cultivation. We then combine previously reported engineering strategies to produce the heterologous sesquiterpenoid patchoulol to high titers from our engineered green cell factories and show these products are possible to produce in under non-sterile conditions. Our work presents a straightforward means to generate C. reinhardtii strains for broader application in bio-processes for the sustainable generation of products.

show abstract

Heterologous expression of phosphite dehydrogenase in the chloroplast or nucleus enables phosphite utilization and genetic selection in Picochlorum spp.

Cited by 10 publications

References 42 publications

The synthetic future of algal genomes

The synthetic future of algal genomes

Combinatorial Engineering Enables Photoautotrophic Growth in High Cell Density Phosphite-Buffered Media to Support Engineered Chlamydomonas reinhardtii Bio-Production Concepts

Combinatorial engineering for photoautotrophic production of recombinant products from the green microalga Chlamydomonas reinhardtii

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