Biodiesel from microalgae: Ways for increasing the effectiveness of lipid accumulation by genetic engineering methods

Korkhovoy, V.I.; Blume, Ya. B.

doi:10.3103/s0095452713060030

Cited by 6 publications

(5 citation statements)

References 87 publications

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“…The nitrogen-limited conditions consistently generated higher fatty acid peak areas than in nitrogen-replete conditions. This is in agreement with other studies indicating increased TAG content in Cr cells with nitrogen-limitation. − …”

Section: Resultssupporting

confidence: 93%

“…This problem was overcome by Ibanez et al by stimulating a known metabolic change in Saccharomyces cerevisiae cells and monitoring the change in lipid content by mass spectrometry at both the “bulk” and single cell level . Employing the same concept, Cr cells were deprived of nitrogen to induce the production of triacylglycerides (TAGs), lipid droplets, and other fatty acid comprised lipids. − Additionally, cells swell in size in response to nitrogen deprivation, which can be measured by the ATOFMS. Cells were monitored by ATOFMS over 4 days of nitrogen deprivation.…”

Section: Resultsmentioning

confidence: 99%

“…In contrast, cells under nitrogen-limited conditions nearly doubled in size over the course of the experiment, growing from 4−5 μm to 7–8 μm in aerodynamic diameter (Figure ). This is due to the accumulation of various lipids under nitrogen-limited conditions. − Small particle noise generated during nebulization creates the APS mode for particles <3 μm. The ATOFMS does not transmit these particles effectively and thus differs from APS measurements in this size range.…”

Section: Resultsmentioning

confidence: 99%

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Online Analysis of Single Cyanobacteria and Algae Cells under Nitrogen-Limited Conditions Using Aerosol Time-of-Flight Mass Spectrometry

et al. 2015

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Metabolomics studies typically perform measurements on populations of whole cells which provide the average representation of a collection of many cells. However, key mechanistic information can be lost using this approach. Investigating chemistry at the single cell level yields a more accurate representation of the diversity of populations within a cell sample; however, this approach has many analytical challenges. In this study, an aerosol time-of-flight mass spectrometer (ATOFMS) was used for rapid analysis of single algae and cyanobacteria cells with diameters ranging from 1 to 8 μm. Cells were aerosolized by nebulization and directly transmitted into the ATOFMS. Whole cells were determined to remain intact inside the instrument through a combination of particle sizing and imaging measurements. Differences in cell populations were observed after perturbing Chlamydomonas reinhardtii cells via nitrogen deprivation. Thousands of single cells were measured over a period of 4 days for nitrogen-replete and nitrogen-limited conditions. A comparison of the single cell mass spectra of the cells sampled under the two conditions revealed an increase in the dipalmitic acid sulfolipid sulfoquinovosyldiacylglycerol (SQDG), a chloroplast membrane lipid, under nitrogen-limited conditions. Single cell peak intensity distributions demonstrate the ability of the ATOFMS to measure metabolic differences of single cells. The ATOFMS provides an unprecedented maximum throughput of 50 Hz, enabling the rapid online measurement of thousands of single cell mass spectra.

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Section: Resultssupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Online Analysis of Single Cyanobacteria and Algae Cells under Nitrogen-Limited Conditions Using Aerosol Time-of-Flight Mass Spectrometry

et al. 2015

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“…Nutrient deprivation is a known route to perturb lipid and metabolite concentrations of ChRe. − Quantitation requires the cell to be fully lysed, the molecular constituents fully extracted, and matrix suppression effects, due to cellular media or from cellular metabolites, to be normalized. − Collection of LVC-electrosprayed material after single cell droplet ejection and exposure to LVC solvent indicate that cells are efficiently lysed in the system (SI Figure S-5). Further, signal levels from CC125 were not significantly lower than CC503 (wall-less mutant), indicating that CC125 cells are lysed even with the presence of a cell wall (SI Figure S-6).…”

Section: Results and Discussionmentioning

confidence: 99%

Rapid, Untargeted Chemical Profiling of Single Cells in Their Native Environment

2019

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We report a method that enables untargeted, high throughput, and quantitative mass spectrometric analysis of single cells from cell suspension without needing additional sample preparation procedures (e.g., molecular tagging) through the combination of single-cell printer technology and liquid vortex capture−mass spectrometry (SCP-LVC-MS). The operating principle behind the SCP-LVC-MS technology is single cell isolation via small droplet piezoelectric ejection followed by capture of the droplet into an LVC-MS sampling probe. Once exposed to an appropriate solvent, the cell is lysed, extracted, and analyzed by MS. The SCP-LVC-MS approach was validated by measuring the lipid composition of microalgae, Chlamydomonas reinhardtii (ChRe) and Euglena gracilis (EuGr), and HeLa cells in their native growth media. Numerous diacylglyceryltrimethylhomo-Ser (DGTS), phosphatidylcholine (PC), monogalactosyldiacylglycerol (MGDG), and digalactosyldiacylglycerol (DGDG) lipids were observed in single cells. Continuous solvent flow ensures that cells are analyzed rapidly, and no signal carryover between cells is observed. ChRe and EuGr microalgae mixed together in the same solution were differentiated cell-by-cell in real-time based on differences between levels of diacylglyceryltrimethylhomo-Ser (DGTS) and phosphatidylcholine (PC) lipids measured in each cell. Several DGTS lipids present in ChRe were quantified with single-cell resolution by normalizing to a DGTS(32:0) internal standard added to the LVC probe solvent during analysis. Quantitative peak areas were validated by comparing to bulk lipid extracts. Lastly, peak area distributions comprised of hundreds of cells were compared for ChRe after 5 days of nitrogen-limited and normal growth conditions, which show clear differences and the ability to resolve cellular population differences with single-cell resolution.

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“…In microalgae, glycerol-3-phosphate dehydrogenase (G3PDH) catalyzes the conversion of dihydroxyacetone phosphate (DHAP) to glycerol-3-phosphate in the cytosol, followed by glycerol-3-phosphate acyltransferase (GPAT) which catalyzes the conversion of glycerol-3-phosphate to lysophosphatidic acid, lysophosphatidic acid acyltransferase (LPAAT) the conversion of lysophosphatidic acid to phosphatidic acid, phosphatidic acid phosphatase (PAP) the conversion of phosphatidic acid to diacylglycerol, and finally diacylglycerol acyltransferase (DGAT) the conversion of diacylglycerol to triacylglycerol (TAG) in the endoplasmic reticulum ( Korkhovoy and Blume, 2013 ; Figure 1 ). Hsieh et al adopted a multiple gene expression strategy to elevate the lipid accumulation of microalgae ( Hsieh et al, 2012 ).…”

Section: Construction Of High-efficient Lipid Producing Microalgaementioning

confidence: 99%

Enhancing microalgal lipid accumulation for biofuel production

Zhu

Sun²,

Fei³

et al. 2022

Front. Microbiol.

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Microalgae have high lipid accumulation capacity, high growth rate and high photosynthetic efficiency which are considered as one of the most promising alternative sustainable feedstocks for producing lipid-based biofuels. However, commercialization feasibility of microalgal biofuel production is still conditioned to the high production cost. Enhancement of lipid accumulation in microalgae play a significant role in boosting the economics of biofuel production based on microalgal lipid. The major challenge of enhancing microalgal lipid accumulation lies in overcoming the trade-off between microalgal cell growth and lipid accumulation. Substantial approaches including genetic modifications of microalgal strains by metabolic engineering and process regulations of microalgae cultivation by integrating multiple optimization strategies widely applied in industrial microbiology have been investigated. In the present review, we critically discuss recent trends in the application of multiple molecular strategies to construct high performance microalgal strains by metabolic engineering and synergistic strategies of process optimization and stress operation to enhance microalgal lipid accumulation for biofuel production. Additionally, this review aims to emphasize the opportunities and challenges regarding scaled application of the strategic integration and its viability to make microalgal biofuel production a commercial reality in the near future.

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Biodiesel from microalgae: Ways for increasing the effectiveness of lipid accumulation by genetic engineering methods

Cited by 6 publications

References 87 publications

Online Analysis of Single Cyanobacteria and Algae Cells under Nitrogen-Limited Conditions Using Aerosol Time-of-Flight Mass Spectrometry

Online Analysis of Single Cyanobacteria and Algae Cells under Nitrogen-Limited Conditions Using Aerosol Time-of-Flight Mass Spectrometry

Rapid, Untargeted Chemical Profiling of Single Cells in Their Native Environment

Enhancing microalgal lipid accumulation for biofuel production

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