Microalgae as an Efficient Vehicle for the Production and Targeted Delivery of Therapeutic Glycoproteins against SARS-CoV-2 Variants DOI Creative Commons
Jaber Dehghani, Ali Movafeghi, Elodie Mathieu‐Rivet

et al.

Marine Drugs, Journal Year: 2022, Volume and Issue: 20(11), P. 657 - 657

Published: Oct. 23, 2022

Severe acute respiratory syndrome–Coronavirus 2 (SARS-CoV-2) can infect various human organs, including the respiratory, circulatory, nervous, and gastrointestinal ones. The virus is internalized into cells by binding to angiotensin-converting enzyme (ACE2) receptor through its spike protein (S-glycoprotein). As S-glycoprotein required for attachment entry target cells, it primary mediator of SARS-CoV-2 infectivity. Currently, this glycoprotein has received considerable attention as a key component development antiviral vaccines or biologics against SARS-CoV-2. Moreover, since ACE2 constitutes main route virus, soluble form could be considered promising approach treatment coronavirus disease 2019 infection (COVID-19). Both are highly glycosylated molecules containing 22 7 consensus N-glycosylation sites, respectively. N-glycan structures attached these specific sites folding, conformation, recycling, biological activity both glycoproteins. Thus far, recombinant have been produced primarily in mammalian which an expensive process. Therefore, benefiting from cheaper cell-based biofactory would good value added cost-effective biopharmaceuticals directed COVID-19. To end, efficient synthesis machinery ability properly impose post-translational modifications make microalgae eco-friendly platform production pharmaceutical Notably, several (e.g., Chlamydomonas reinhardtii, Dunaliella bardawil, Chlorella species) already approved U.S. Food Drug Administration (FDA) safe food. Because microalgal contain rigid cell wall that act natural encapsulation protect proteins aggressive environment stomach, feature used rapid edible targeted delivery treatment/inhibition Herein, we reviewed pathogenesis mechanism then highlighted potential COVID-19 infection.

Language: Английский

Harnessing genetic engineering to drive economic bioproduct production in algae DOI Creative Commons
Abhishek Gupta, Kalisa Kang, Ruchi Pathania

et al.

Frontiers in Bioengineering and Biotechnology, Journal Year: 2024, Volume and Issue: 12

Published: Jan. 29, 2024

Our reliance on agriculture for sustenance, healthcare, and resources has been essential since the dawn of civilization. However, traditional agricultural practices are no longer adequate to meet demands a burgeoning population amidst climate-driven challenges. Microalgae emerge as beacon hope, offering sustainable renewable source food, animal feed, energy. Their rapid growth rates, adaptability non-arable land non-potable water, diverse bioproduct range, encompassing biofuels nutraceuticals, position them cornerstone future resource management. Furthermore, microalgae’s ability capture carbon aligns with environmental conservation goals. While microalgae offers significant benefits, obstacles in cost-effective biomass production persist, which curtails broader application. This review examines compared other host platforms, highlighting current innovative approaches aimed at overcoming existing barriers. These include range techniques, from gene editing, synthetic promoters, mutagenesis selective breeding metabolic engineering through transcription factors.

Language: Английский

Citations

20
Introns mediate post-transcriptional enhancement of nuclear gene expression in the green microalga Chlamydomonas reinhardtii DOI Creative Commons
Thomas Baier, Nick Jacobebbinghaus, Alexander Einhaus

et al.

PLoS Genetics, Journal Year: 2020, Volume and Issue: 16(7), P. e1008944 - e1008944

Published: July 30, 2020

Efficient nuclear transgene expression in the green microalga Chlamydomonas reinhardtii is generally hindered by low transcription rates. Introns can increase transcript abundance a process called Intron-Mediated Enhancement (IME) this alga and has been broadly observed other eukaryotes. However, mechanisms of IME microalgae are poorly understood. Here, we identified 33 native introns from highly expressed genes C. selected transcriptome studies as well 13 non-native introns. We investigated their capacities probed mechanism action modification splice sites, internal sequence motifs, position within transgenes. Several were found to elicit strong be applicable different constructs. determined that exclusively occurs transcribed ORFs regardless promoter not induced traditional enhancers transcription. Our results elucidate some mechanistic details reinhardtii, which similar those higher plants yet underly distinctly induction processes. findings narrow focus targets responsible for algal provides evidence underestimated regulators gene expression.

Language: Английский

Citations

77
Real-time monitoring of subcellular H2O2 distribution in Chlamydomonas reinhardtii DOI Creative Commons
Justus Niemeyer, David Scheuring, Julian Oestreicher

et al.

The Plant Cell, Journal Year: 2021, Volume and Issue: 33(9), P. 2935 - 2949

Published: June 30, 2021

Hydrogen peroxide (H2O2) is recognized as an important signaling molecule in plants. We sought to establish a genetically encoded, fluorescent H2O2 sensor that allows monitoring all major subcompartments of Chlamydomonas cell. To this end, we used the Modular Cloning toolbox target hypersensitive reduction–oxidation sensitive green protein2-Tsa2ΔCR cytosol, nucleus, mitochondrial matrix, chloroplast stroma, thylakoid lumen, and endoplasmic reticulum (ER). The was functional compartments, except for ER where it fully oxidized. Employing our novel sensors, show produced by photosynthetic linear electron transport (PET) stroma leaks into cytosol but only reaches other subcellular compartments if under nonphysiological conditions. Furthermore, heat-stressed cells, cytosolic levels closely mirror temperature up- downshifts are independent from PET. Heat stress led similar nucleus and, more mildly, mitochondria not chloroplast. Our results thus suggest establishment steep intracellular gradients normal physiological conditions with limited diffusion compartments. anticipate these sensors will greatly facilitate future investigations biology plant cells.

Language: Английский

Citations

67
Metabolic Engineering for Efficient Ketocarotenoid Accumulation in the Green Microalga Chlamydomonas reinhardtii DOI
Sofia Amendola, Jacob Sebastian Kneip, Florian Meyer

et al.

ACS Synthetic Biology, Journal Year: 2023, Volume and Issue: 12(3), P. 820 - 831

Published: Feb. 23, 2023

Astaxanthin is a valuable ketocarotenoid with various pharmaceutical and nutraceutical applications. Green microalgae harbor natural capacities for pigment accumulation due to their 2-C-methyl-d-erythritol 4-phosphate (MEP) pathway. Recently, redesigned ß-carotene ketolase (BKT) was found enable in the model microalga Chlamydomonas reinhardtii, transformants exhibited reduced photoinhibition under high-light. Here, systematic screening by synthetic transgene design of carotenoid pathway enzymes overexpression from nuclear genome identified phytoene synthase (PSY/crtB) as bottleneck C. reinhardtii. Increased hydroxylase (CHYB) activity be essential engineered astaxanthin accumulation. A combined BKT, crtB, CHYB expression strategy resulted volumetric production 9.5 ± 0.3 mg L–1 (4.5 0.1 g–1 CDW) mixotrophic 23.5 (1.09 h–1) high cell density conditions, 4-fold increase compared previous reports This work presents investigation bottlenecks reinhardtii phototrophic green factory competitive use industrial biotechnology.

Language: Английский

Citations

27
Overexpression of Sedoheptulose-1,7-Bisphosphatase Enhances Photosynthesis in Chlamydomonas reinhardtii and Has No Effect on the Abundance of Other Calvin-Benson Cycle Enzymes DOI Creative Commons

Alexander Hammel,

Frederik Sommer, David Zimmer

et al.

Frontiers in Plant Science, Journal Year: 2020, Volume and Issue: 11

Published: June 23, 2020

The productivity of plants and microalgae needs to be increased feed the growing world population promote development a low-carbon economy. This goal can achieved by improving photosynthesis via genetic engineering. In this study, we have employed Modular Cloning strategy overexpress Calvin-Benson cycle (CBC) enzyme sedoheptulose-1,7 bisphosphatase (SBP1) up 3-fold in unicellular green alga Chlamydomonas reinhardtii. protein derived from nuclear transgene represented ~0.3% total cell protein. Photosynthetic rate growth were significantly SBP1-overexpressing lines under high-light elevated CO2 conditions. Absolute quantification abundance all other CBC enzymes QconCAT approach revealed no consistent differences between recipient strain. analysis also that eleven represent 11.9% Chlamydomonas. Here range concentrations turned out much larger than estimated earlier, with 128-fold difference most abundant (rbcL) least (triose phosphate isomerase). Accordingly, intermediates are often but not always higher binding site for which they act as substrates. highest substrate ratios might good candidates overexpression subsequent engineering steps.

Language: Английский

Citations

57
High cell density cultivation enables efficient and sustainable recombinant polyamine production in the microalga Chlamydomonas reinhardtii DOI
Robert A. Freudenberg, Thomas Baier, Alexander Einhaus

et al.

Bioresource Technology, Journal Year: 2020, Volume and Issue: 323, P. 124542 - 124542

Published: Dec. 16, 2020

Language: Английский

Citations

55
Rational Promoter Engineering Enables Robust Terpene Production in Microalgae DOI
Alexander Einhaus, Thomas Baier,

Marian Rosenstengel

et al.

ACS Synthetic Biology, Journal Year: 2021, Volume and Issue: 10(4), P. 847 - 856

Published: March 25, 2021

Microalgal biotechnology promises sustainable light-driven production of valuable bioproducts and addresses urgent demands to attain a economy. However, unfold its full potential as platform for biotechnology, new powerful tools nuclear engineering need be established. Chlamydomonas reinhardtii, the model microalgal synthetic biology genetic has already been used produce various bioproducts. Nevertheless, low transgene titers, lack potent expression elements, sparse comparative evaluation prevents further development C. reinhardtii biotechnological host. By systematically evaluating existing elements combined with rational promoter engineering, we established novel improved standardized application tools, unveiled an synergism between PSAD 5′ UTR corresponding chloroplast targeting peptide. Promoter strategies, implemented in newly designed algal promoter, increased sesquiterpene (E)-α-bisabolene by 18-fold compared native version 4-fold commonly elements. Our results improve microalgae display significant step toward establishing green cell-factory.

Language: Английский

Citations

55
Engineering a powerful green cell factory for robust photoautotrophic diterpenoid production DOI
Alexander Einhaus,

Jasmin Steube,

Robert A. Freudenberg

et al.

Metabolic Engineering, Journal Year: 2022, Volume and Issue: 73, P. 82 - 90

Published: June 16, 2022

Language: Английский

Citations

34
Engineered production of isoprene from the model green microalga Chlamydomonas reinhardtii DOI Creative Commons
Razan Z. Yahya, Gordon B. Wellman, Sebastian Overmans

et al.

Metabolic Engineering Communications, Journal Year: 2023, Volume and Issue: 16, P. e00221 - e00221

Published: March 23, 2023

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

Language: Английский

Citations

20
An oral delivery system for controlling white spot syndrome virus infection in shrimp using transgenic microalgae DOI

Asama Kiataramgul,

Sugunya Maneenin,

Saul Purton

et al.

Aquaculture, Journal Year: 2020, Volume and Issue: 521, P. 735022 - 735022

Published: Jan. 27, 2020

Language: Английский

Citations

48