A Chloroplast-Localised Fluorescent Protein Enhances the Photosynthetic Action Spectrum in Green Algae

Microorganisms, Journal Year: 2022, Volume and Issue: 10(9), P. 1770 - 1770

Published: Sept. 1, 2022

Green microalgae are important sources of natural products and attractive cell factories for manufacturing high-value such as recombinant proteins. Increasing scales production must address the bottleneck providing sufficient light energy photosynthesis. Enhancing photosynthetic action spectrum green algae to improve utilisation yellow would provide additional Here, we evaluated Katushka fluorescent protein, which converts photons red photons, drive photosynthesis growth when expressed in Chlamydomonas reinhardtii chloroplasts. Transplastomic expressing a codon-optimised gene accumulated active was detected by excitation with light. Removal chlorophyll from cells, captures led increased fluorescence. In light, emission oxygen evolution growth. Utilisation transplastomic cells deficient photons. These results showed that simple effective light-capturing device enhanced C. reinhardtii.

Language: Английский

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Accelerating Chloroplast Engineering: A New System for Rapid Generation of Marker-Free Transplastomic Lines of Chlamydomonas reinhardtii

Microorganisms, Journal Year: 2023, Volume and Issue: 11(8), P. 1967 - 1967

Published: July 31, 2023

'Marker-free' strategies for creating transgenic microorganisms avoid the issue of potential transmission antibiotic resistance genes to other microorganisms. An already-established strategy engineering chloroplast genome (=plastome) green microalga Chlamydomonas reinhardtii involves restoration photosynthetic function using a recipient strain carrying plastome mutation in key photosynthesis gene. Selection transformant colonies is carried out on minimal media, such that only those cells which mutated gene has been replaced with wild-type copy DNA are capable phototrophic growth. However, this approach can suffer from issues efficiency due slow growth C. media and die-back untransformed lawn when mutant strains limited photosensitivity phenotype. Furthermore, rescue tended rely existing mutants not necessarily ideal transformation targeted transgene insertion: Mutants point mutations easily revert, deletions do extend intended insertion site give rise sub-population rescued lines lack transgene. In order improve accelerate pipeline reinhardtii, we have created novel line, HNT6, an engineered deletion exon 3 psaA, encodes one core subunits photosystem I (PSI). Such PSI highly light-sensitive allowing faster recovery by selecting light-tolerance acetate-containing rather than media. The extends upstream psaA-3 serves as neutral locus insertion, thereby ensuring all recovered transformants containing We demonstrate application HNT6 luciferase reporter.

Language: Английский

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Over-expression of a cyanobacterial gene for 1-deoxy-d-xylulose-5-phosphate synthase in the chloroplast of Chlamydomonas reinhardtii perturbs chlorophyll: carotenoid ratios

Journal of King Saud University - Science, Journal Year: 2022, Volume and Issue: 34(6), P. 102141 - 102141

Published: June 2, 2022

Terpenoids are a diverse class of naturally occurring compounds consisting more than 50,000 structurally different molecules and found in all living organisms. Many terpenoid compounds, particular those isolated from plants, have applications various commercial sectors including medicine, agriculture cosmetics. However, these high value terpenoids produced relatively small quantities their natural hosts chemical synthesis for large scale production is costly complicated. Therefore, there much focus on producing novel biological using metabolic engineering technologies. As photosynthetic system, the unicellular green alga C. reinhardtii interest as most well-studied model with well-established molecular tools genetic manipulation. direct manipulation biosynthetic pathways necessitates thorough understanding basic metabolism. To gain better methylerythritol phosphate (MEP) pathway that leads to biosynthesis chloroplast reinhardtii, hence this study has investigated effect over-expressing 1-deoxy-d-xylulose-5-phosphate synthase (DXS) plastidic downstream terpenoids. We marker-free transformants lines express an additional cyanobacterial gene DXS. The analysis content transgenic line demonstrates overexpressing DXS resulted two-fold decrease chlorophyll levels while carotenoid showed variable changes: zeaxanthin antherxanthin increased several-fold, lutein dropped approximately half, but β-carotene violaxanthin did not show significant change.

Language: Английский

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Three Parts of the Plant Genome: On the Way to Success in the Production of Recombinant Proteins

Plants, Journal Year: 2022, Volume and Issue: 12(1), P. 38 - 38

Published: Dec. 21, 2022

Recombinant proteins are the most important product of current industrial biotechnology. They indispensable in medicine (for diagnostics and treatment), food chemical industries, research. Plant cells combine advantages eukaryotic protein production system with simplicity efficacy bacterial one. The use plants for recombinant is an economically promising area that has emerged as alternative to traditional approaches. This review discusses plant systems expression using nuclear, plastid, mitochondrial genomes. Possibilities, problems, prospects modifications three parts genome light obtaining producer examined. Examples successful nuclear platform various biopharmaceuticals, veterinary drugs, technologically described, examples a high yield upon modification chloroplast genome. Potential utility mitochondria its over nucleus chloroplasts substantiated. Although these opportunities have not yet been exploited, potential

Language: Английский

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Microalgae as expression systems for recombinant protein production

Revista Tecnología en Marcha, Journal Year: 2024, Volume and Issue: unknown

Published: Nov. 29, 2024

In the field of biotechnology, recombinant proteins have revolutionized many industries, including pharmaceuticals, agriculture, and bioenergy. By producing high-value in heterologous hosts, cell factories may offer a more efficient, cost-effective, scalable, environmentally friendly solution to traditional protein production extraction methods, which can be highly laborious resource intensive. Microalgae emerged as attractive hosts due their Generally Recognized Safe (GRAS) status, versatile metabolism, genetic diversity between species, ease cultivation scale-up, general cost-effectiveness. For engineering, capability for complex synthesis post-translational modifications transformation comparison with chasses outside category make microalgae an advantageous on fronts. transformed enable efficient expression, most commonly nucleus chloroplast, each specific advantages limitations. The present literature review compiles some techniques, features, latest advances related microalgae, exploring different techniques Recombinant is only one processes that become sustainable by using platform.

Language: Английский

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