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Crypthecodinium cohnii strain:ATCC 30556

ABSTRACT: Crypthecodinium cohnii Raw RNA-sequencing reads

PROVIDER: PRJNA318728 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR3400310_1.fastq.gz	Fastqsanger.gz
	SRR3400310_2.fastq.gz	Fastqsanger.gz
	SRR3400311_1.fastq.gz	Fastqsanger.gz
	SRR3400311_2.fastq.gz	Fastqsanger.gz
	SRR3400312_1.fastq.gz	Fastqsanger.gz

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Rewiring the metabolic network to increase docosahexaenoic acid productivity in Crypthecodinium cohnii by fermentation supernatant-based adaptive laboratory evolution

Project description:To overcome the inhibition caused by the fermentation supernatant in the late fermentation stage of docosahexaenoic acid (DHA)-producing Crypthecodinium cohnii, fermentation supernatant-based adaptive laboratory evolution (FS-ALE) was conducted. The cell growth and DHA productivity of the evolved strain (FS280) obtained after 280 adaptive cycles corresponding to 840 days of evolution were increased by 161.87% and 311.23%, respectively, at 72 h under stress conditions and increased by 19.87% and 51.79% without any stress compared with the starting strain, demonstrating the effectiveness of FS-ALE.

2021-11-27 | PXD030028 |

Separation of Heterotrophic Microalgae <i>Crypthecodinium cohnii</i> by Dielectrophoresis.

Project description:Microalgae constitute an abundant source of poly-unsaturated fatty acids which are applied in various biotechnological fields such as pharmaceuticals and food supplement. Separating microalgae cells with respect to their lipid content would establish a relevant at-line analytical technique. The present study demonstrates an electrical approach for the separation of the lipid-producing microalgae Crypthecodinium cohnii using the effect of dielectrophoresis (DEP) in a microfluidic flow cell. Microalgae were cultivated for 8 days, while cell growth was characterized by optical density, dry cell weight, glucose concentration and lipid content via fluorescence microscopy. The size distribution of cells during cultivation was thoroughly investigated, since the DEP force scales with cell volume, but also depends on lipid content via cell electrophysiological constants. Thus, the challenge was to deconvolute one separation effect from the other, while the electrical cell constants of C. cohnii are not known yet. The DEP-dependent separation was realized by slanted top-bottom electrodes with the flowing cell suspension between them. Turning on the voltage deflected the cells from their initial path as determined by the streaming and thus changed their direction of flow. The separation efficiency of DEP was tested for various electrical field strengths and its performance was determined by quantitative analysis of optical and fluorescence videos. It could be shown for all size groups that the most lipid-containing cells were always subject to DEP separation and that the method is thus not only suitable for process analysis, but also for strain selection of the most productive cell lines.

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Genetic Engineering of Crypthecodinium cohnii to Increase Growth and Lipid Accumulation.

Project description:In this study, we evaluated suitable selected markers and optimized transformation protocols to develop a new genetic transformation methodology for DHA-producing Crypthecodinium cohnii. Additionally, ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO), potentially involved in CO2 fixation under autotrophic conditions, was selected as the target for construction of a gene knockdown mutant. Our results show that the constructs were successfully inserted into the C. cohnii chromosome by homologous recombination. Comparative analysis showed that deletion of the RuBisCO gene promoted cell growth and increased the lipid content of C. cohnii under heterotrophic conditions compared with those of the wild-type. The liquid chromatography-mass spectrometry (LC-MS) based metabolomic analysis showed that the metabolites involved in energy metabolism were upregulated, suggesting that the deletion of the RuBisCO gene may contribute to the re-direction of more carbon or energy toward growth and lipid accumulation under heterotrophic conditions.

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