Journal
ALGAL RESEARCH-BIOMASS BIOFUELS AND BIOPRODUCTS
Volume 58, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.algal.2021.102385
Keywords
Nannochloropsis; CRISPR/Cas9; Carbohydrate; Glucan synthase; Transglycosylase
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Funding
- Department of Energy's Bio-energy Technologies Office [DEEE0007089]
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The study knocked out two key enzymes responsible for chrysolaminarin synthesis in Nannochloropsis species using CRISPR/Cas9 technology, resulting in a significant decrease in soluble carbohydrate accumulation following nitrogen starvation.
Nannochloropsis species have garnered significant interest for biofuel production due to their ability to accumulate high levels of triacylglycerols (TAGs), especially following nitrogen-starvation. However, the first response to nutrient starvation is the synthesis of chrysolaminarin, a soluble beta-1,3-glucan with beta-1,6-branching. We employ CRISPR/Cas9 to knock out two key enzymes responsible for the synthesis of this oligosaccharide: a beta-glucan synthase (BGS) gene putatively responsible for the glucose beta-1,3-linkages, and a transglycosylase (TGS) which putatively catalyzes beta-1,6-branching. Analysis of the biomass from the generated mutants confirmed an similar to 5-fold decrease in the accumulation of soluble carbohydrate following nitrogen starvation, without an observed growth defect in a diel light-cycling regime compared to CRISPR-expressing controls.
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