Screening of oleaginous algal strains from <i>Chlamydomonas reinhardtii</i> mutant libraries via density gradient centrifugation

Sung, Young Joon; Choi, Hong Il; Lee, Jeong Seop; Hong, Min Eui; Sim, Sang Jun

doi:10.1002/bit.27149

Cited by 13 publications

(5 citation statements)

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“…In this regard, it is likely that lipid accumulation might account for the increasing trend in cell density we observed in TAP-NS. Nutrient starvation leads to a slower growth rate and eventually growth arrest, cell division arrest or cell death in microalgae [ 35 , 36 ]. During nitrogen limitation, cells rearrange their intracellular macromolecular pools, leading to mobilization of carbon to neutral lipid synthesis [ 36 ].…”

Section: Resultsmentioning

confidence: 99%

“…In addition, cc5373-sta6 subjected to TAP-NS displayed a gradual increase in cell density during the six-day period. The cc5373-sta6 has been reported to produce significantly higher levels of lipid accumulation than controls under nitrogen deprivation [ 10 , 36 ]. The increased cellular reactive oxygen species (ROS) play a dual role in signaling lipid biosynthesis and inducing autophagy to recycle specific cellular components [ 10 , 39 ].…”

Section: Resultsmentioning

confidence: 99%

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Comparison of the effects of nitrogen-, sulfur- and combined nitrogen- and sulfur-deprivations on cell growth, lipid bodies and gene expressions in Chlamydomonas reinhardtii cc5373-sta6

Gonzalez,

Ynalvez

2023

BMC Biotechnol

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Background Biofuel research that aims to optimize growth conditions in microalgae is critically important. Chlamydomonas reinhardtii is a green microalga that offers advantages for biofuel production research. This study compares the effects of nitrogen-, sulfur-, and nitrogen and sulfur- deprivations on the C. reinhardtii starchless mutant cc5373-sta6. Specifically, it compares growth, lipid body accumulation, and expression levels of acetyl-CoA carboxylase (ACC) and phosphoenolpyruvate carboxylase (PEPC). Results Among nutrient-deprived cells, TAP-S cells showed significantly higher total chlorophyll, cell density, and protein content at day 6 (p < 0.05). Confocal analysis showed a significantly higher number of lipid bodies in cells subjected to nutrient deprivation than in the control over the course of six days; N deprivation for six days significantly increased the size of lipid bodies (p < 0.01). In comparison with the control, significantly higher ACC expression was observed after 8 and 24 h of NS deprivation and only after 24 h with N deprivation. On the other hand, ACC and PEPC expression at 8 and 24 h of S deprivation was not significantly different from that in the control. A significantly lower PEPC expression was observed after 8 h of N and NS deprivation (p < 0.01), but a significantly higher PEPC expression was observed after 24 h (p < 0.01). Conclusions Based on our findings, it would be optimum to cultivate cc5373-sta6 cells in nutrient deprived conditions (-N, -S or –NS) for four days; whereby there is cell growth, and both a high number of lipid bodies and a larger size of lipid bodies produced.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Comparison of the effects of nitrogen-, sulfur- and combined nitrogen- and sulfur-deprivations on cell growth, lipid bodies and gene expressions in Chlamydomonas reinhardtii cc5373-sta6

Gonzalez,

Ynalvez

2023

BMC Biotechnol

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“…Percoll density centrifugation selectively sorts the mutant cells based on their relative densities [55]. The low-density cells are enriched during the procedure, as they are the ones with high lipid content, since lipids have lower density (0.9 g/cm 3 ) than other cellular biomolecules, such as carbohydrates, proteins, and nucleotides [56].…”

Section: Screening Methods For Selecting Promising Mutantsmentioning

confidence: 99%

“…Microalgae are considered a potential large-scale sink for CO 2 sequestration from flue gases, which contain 12-15% CO 2 [56,57]. However, most of the microalgal strains can only survive up to 5% CO 2 , hence development of high tolerance CO 2 strains is crucial [100][101][102].…”

Section: Microalga Mutantsmentioning

confidence: 99%

Harnessing the Power of Mutagenesis and Adaptive Laboratory Evolution for High Lipid Production by Oleaginous Microalgae and Yeasts

2020

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Oleaginous microalgae and yeasts represent promising candidates for large-scale production of lipids, which can be utilized for production of drop-in biofuels, nutraceuticals, pigments, and cosmetics. However, low lipid productivity and costly downstream processing continue to hamper the commercial deployment of oleaginous microorganisms. Strain improvement can play an essential role in the development of such industrial microorganisms by increasing lipid production and hence reducing production costs. The main means of strain improvement are random mutagenesis, adaptive laboratory evolution (ALE), and rational genetic engineering. Among these, random mutagenesis and ALE are straight forward, low-cost, and do not require thorough knowledge of the microorganism’s genetic composition. This paper reviews available mutagenesis and ALE techniques and screening methods to effectively select for oleaginous microalgae and yeasts with enhanced lipid yield and understand the alterations caused to metabolic pathways, which could subsequently serve as the basis for further targeted genetic engineering.

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“…FAME was detected using the gas chromatography-flame ionization detection (GC-FID) method and a DB-23 column (Agilent, USA). The detailed experimental conditions for detection of FAME has been reported elsewhere [30]. The amount of FAME was quantified via the correlation between peak area and concentration of internal standard.…”

Section: Lipid Extraction and Analysis Of Algal Cellsmentioning

confidence: 99%

Outdoor cultivation of microalgae in a coal-fired power plant for conversion of flue gas CO2 into microalgal direct combustion fuels

Sung

Lee

Yoon

et al. 2020

Syst Microbiol and Biomanuf

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Microalgae have piqued renewed interest as a sustainable biofuel feedstock owing to their high CO 2 conversion efficiency. However, the major limitation of microalga-based biofuel production is low productivity. In this study, CO 2 in flue gas emitted from the coal-fired power plants was fixed through mass microalgal cultivation using only sunlight as an energy source. To minimize the cost and energy required to supply the flue gas and efficiently utilize the microalgal biomass, a polycarbonate (PC) greenhouse and polymeric photobioreactors were installed near the power plant stack. Four different microalgal strains (Chlamydomonas reinhardtii, Chlorella sorokiniana, Neochloris oleoabundans, and Neochloris oleoabundans #13) were subjected to semi-continuous culturing for 1 month. The maximum biomass productivity was achieved by the N. oleoabundans #13 strain (0.703 g L −1 day −1). Additionally, polymerase chain reaction analysis revealed that the individual microalgal culture was not cross-contaminated with other microalgal cultures in this cultivation system, owing to the structural properties of photobioreactor comprising individual modules. The lipid content and calorific productivity of N. oleoabundans #13 biomass were 45.70% and 3.553 kJ L −1 day −1 , respectively, which indicate satisfactory performance of biomass as a direct combustion fuel. The CO 2 fixation rate, which was calculated based on the carbon content in the biomass, was 0.309 g CO 2 L −1 day −1. Therefore, large amounts of CO 2 can be reduced using the large-scale microalgal cultivation system, which enables efficient biological CO 2 conversion and maximizes microalgal biomass utilization.

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Screening of oleaginous algal strains from Chlamydomonas reinhardtii mutant libraries via density gradient centrifugation

Cited by 13 publications

References 48 publications

Comparison of the effects of nitrogen-, sulfur- and combined nitrogen- and sulfur-deprivations on cell growth, lipid bodies and gene expressions in Chlamydomonas reinhardtii cc5373-sta6

Comparison of the effects of nitrogen-, sulfur- and combined nitrogen- and sulfur-deprivations on cell growth, lipid bodies and gene expressions in Chlamydomonas reinhardtii cc5373-sta6

Harnessing the Power of Mutagenesis and Adaptive Laboratory Evolution for High Lipid Production by Oleaginous Microalgae and Yeasts

Outdoor cultivation of microalgae in a coal-fired power plant for conversion of flue gas CO2 into microalgal direct combustion fuels

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