• Journal of Microbiology and Biotechnology
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• v.30 no.1
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• pp.136-145
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• 2020

Chlorella sp. HS2, which previously showed excellent performance in phototrophic cultivation and has tolerance for wide ranges of salinity, pH, and temperature, was cultivated heterotrophically. However, this conventional medium has been newly optimized based on a composition analysis using elemental analysis and ICP-OES. In addition, in order to maintain a favorable dissolved oxygen level, stepwise elevation of revolutions per minute was adopted. These optimizations led to 40 and 13% increases in the biomass and lipid productivity, respectively (7.0 and 2.25 g l^-1d^-1 each). To increase the lipid content even further, 12 h heat shock at 50℃ was applied and this enhanced the biomass and lipid productivity up to 4 and 17% respectively (7.3 and 2.64 g l^-1d^-1, each) relative to the optimized conditions above, and the values were 17 and 14% higher than ordinary lipid-accumulating N-limitation (6.2 and 2.31 g l^-1d^-1). On this basis, heat shock was successfully adopted in novel Chlorella sp. HS2 cultivation as a lipid inducer for the first time. Considering its fast and cost-effective characteristics, heat shock will enhance the overall microalgal biofuel production process.

• Journal of Microbiology and Biotechnology
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• v.33 no.3
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• pp.310-318
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• 2023

Microalgae are attracting much attention as promising, eco-friendly producers of bioenergy due to their fast growth, absorption of carbon dioxide from the atmosphere, and production capacity in wastewater and salt water. However, microalgae can only accumulate large quantities of lipid in abiotic stress, which reduces productivity by decreasing cell growth. In this study, the strategy was investigated to increase cell viability and lipid production by overexpressing S-adenosylmethionine (SAM) synthetase (SAMS) in the microalga Chlamydomonas reinhardtii. SAM is a substance that plays an important role in various intracellular biochemical reactions, such as cell proliferation and stress response, and the overexpression of SAMS could allow cells to ithstand the abiotic stress and increase productivity. Compared to wild-type C. reinhardtii, recombinant cells overexpressing SAMS grew 1.56-fold faster and produced 1.51-fold more lipids in a nitrogen-depleted medium. Furthermore, under saline-stress conditions, the survival rate and lipid accumulation were 1.56 and 2.04 times higher in the SAMS-overexpressing strain, respectively. These results suggest that the overexpression of SAMS in recombinant C. reinhardtii has high potential in the industrial-scale production of biofuels and various other high-value-added materials.

• Journal of Microbiology and Biotechnology
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• v.25 no.6
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• pp.910-917
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• 2015

A total of 646 strains, including green algae and diatoms, were isolated from 220 samples to screen microalgae with high lipid productivity (LP). The samples were obtained from nine habitats in Northern Xinjiang, China in June 2013. This study initially identified eight lipidrich strains, namely, Desmodesmus intermedius XJ-498, D. intermedius XJ-145, D. intermedius XJ-99, Monoraphidium pusillum XJ-489, M. dybowskii XJ-435, M. dybowskii XJ-151, Mychonastes homosphaera XJ-488, and Podohedriella falcata XJ-176, based on 18S rDNA sequencing. The strains were cultured in a photobioreactor for the same period. Results showed that the specific growth rate (day^-1) of M. pusillum XJ-489 was the highest (1.14 ± 0.06), and the biomass concentration (g/l) of D. intermedius XJ-99 was the highest (2.84 ± 0.3). Futhermore, the lipid content (%) of M. dybowskii XJ-151 was the highest (33.5 ± 4.38), and the lipid productivity (mg l^-1 day^-1) of My. homosphaera XJ-488 was the highest (86.41 ± 9.04). C16 to C18 accounted for 86% to 98% of the total lipid, and the biodiesel qualities of the selected algae corresponded to international standards. This study suggests that My. homosphaera XJ-488, D. intermedius XJ-99, and M. dybowskii XJ-151 are the most potential strains for biodiesel production among all the isolated strains.

• Microbiology and Biotechnology Letters
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• v.49 no.2
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• pp.210-216
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• 2021

This study aimed to enhance ethanol productivity of Saccharomyces cerevisiae through genome editing using CRISPR/CAS9. To increase ethanol productivity, ACC1, ELO1, and OLE1 were overexpressed in S. cerevisiae using the CRISPR/CAS9 system. The strains overexpressing ACC1, ELO1, and OLE1 survived up to 24 h in YPD medium supplemented with 18% ethanol. Moreover, the ethanol yields in strains overexpressing ACC1 (428.18 mg ethanol/g glucose), ELO1 (416.15 mg ethanol/g glucose), and OLE1 (430.55 mg ethanol/g glucose) were higher than those in the control strains (400.26 mg ethanol/g glucose). In conclusion, the overexpression of these genes increased the viability of S. cerevisiae at high ethanol concentrations and the ethanol productivity without suppressing glucose consumption.

• ALGAE
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• v.31 no.3
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• pp.257-266
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• 2016

Nitrogen is one of the most critical nutrients affecting cell growth and biochemical composition of microalgae, ultimately determining the lipid or carbohydrate productivity for biofuels. In order to investigate the effect of nitrogen sources on the cell growth and biochemical composition of the marine microalga Tetraselmis sp., nine different N sources, including NaNO₃, KNO₃, NH₄NO₃, NH₄HCO₃, NH₄Cl, CH₃COONH₄, urea, glycine, and yeast extract were compared at the given concentration of 8.82 mM. Higher biomass concentration was achieved under organic nitrogen sources, such as yeast extract (2.23 g L⁻¹) and glycine (1.62 g L⁻¹), compared to nitrate- (1.45 g L⁻¹) or ammonium-N (0.98 g L⁻¹). All ammonium sources showed an inhibition of cell growth, but accumulated higher lipids, showing a maximum content of 28.3% in ammonium bicarbonate. When Tetraselmis sp. was cultivated using yeast extract, the highest lipid productivity of 36.0 mg L⁻¹ d⁻¹ was achieved, followed by glycine 21.5 mg L⁻¹ d⁻¹ and nitrate 19.9 mg L⁻¹ d⁻¹. Ammonium bicarbonate resulted in the lowest lipid productivity of 14.4 mg L⁻¹ d⁻¹. The major fatty acids in Tetraselmis sp. were palmitic, oleic, linoleic and linolenic acids, regardless of the nutritional compositions, indicating the suitability of this species for biodiesel production.

• Journal of Microbiology and Biotechnology
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• v.23 no.1
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• pp.92-98
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• 2013

Microalgal cultivation using wastewater is now regarded as essential for biodiesel production, as two goals can be achieved simultaneously; that is, nutrient removal efficiency and biomass production. Therefore, this study examined the effects of carbon sources, the N:P ratio, and the hydraulic retention time (HRT) to identify the optimal conditions for nutrient removal efficiency and biomass production. The effluent from a 2nd lagoon was used to cultivate microalgae. Whereas the algal species diversity and lipid content increased with a longer HRT, the algal biomass productivity decreased. Different carbon sources also affected the algal species composition. Diatoms were dominant with an increased pH when bicarbonate was supplied. However, 2% $CO_2$ gas led to a lower pH and the dominance of filamentous green algae with a much lower biomass productivity. Among the experiments, the highest chlorophyll-a concentration and lipid productivity were obtained with the addition of phosphate up to 0.5 mg/l P, since phosphorus was in short supply compared with nitrogen. The N and P removal efficiencies were also higher with a balanced N:P ratio, based on the addition of phosphate. Thus, optimizing the N:P ratio for the dominant algae could be critical in attaining higher algal growth, lipid productivity, and nutrient removal efficiency.

• Journal of Microbiology and Biotechnology
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• v.23 no.9
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• pp.1260-1268
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• 2013

A series of experiments were carried out with three native strains of microalgae to measure growth rates, biomass, and lipid productivities. Scenedesmus sp. IMMTCC-6 had better biomass growth rate and higher lipid production. The growth, lipid accumulation, and carbon dioxide ($CO_2$) consumption rate of Scenedesmus sp. IMMTCC-6 were tested under different NaOH concentrations in modified BBM. The algal strain showed the maximum specific growth rate (0.474/day), biomass productivity (110.9 mg $l^{-1}d^{-1}$), and $CO_2$ consumption rate (208.4 mg $l^{-1}d^{-1}$) with an NaOH concentration of 0.005 M on the $8^{th}$ day of cultivation. These values were 2.03-, 6.89-, and 6.88-fold more than the algal cultures grown in control conditions (having no NaOH and $CO_2$). The $CO_2$ fixing efficiency of the microalga with other alternative carbon sources like $Na_2CO_3$ and $NaHCO_3$ was also investigated and compared. The optimized experimental parameters at shake-flask scale were implemented for scaling up the process in a self-engineered photobioreactor. A significant increase in lipid accumulation (14.23% to 31.74%) by the algal strain from the logarithmic to stationary phases was obtained. The algal lipids were mainly composed of $C_{16}/C_{18}$ fatty acids, and are desirable for biodiesel production. The study suggests that microalga Scenedesmus sp. IMMTCC-6 is an efficient strain for biodiesel production and $CO_2$ biofixation using stripping solution of NaOH in a cyclic process.

• Journal of Microbiology and Biotechnology
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• v.25 no.1
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• pp.109-118
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• 2015

Open raceway ponds are cost-efficient for mass cultivation of microalgae compared with photobioreactors. Although low-cost options like wastewater as nutrient source is studied to overcome the commercialization threshold for biodiesel production from microalgae, a cost analysis on the use of wastewater and other incremental increases in productivity has not been elucidated. We determined the effect of using wastewater and wavelength filters on microalgal productivity. Experimental results were then fitted into a model, and cost analysis was performed in comparison with control raceways. Three different microalgal strains, Chlorella vulgaris AG10032, Chlorella sp. JK2, and Scenedesmus sp. JK10, were tested for nutrient removal under different light wavelengths (blue, green, red, and white) using filters in batch cultivation. Blue wavelength showed an average of 27% higher nutrient removal and at least 42% higher chemical oxygen demand removal compared with white light. Naturally, the specific growth rate of microalgae cultivated under blue wavelength was on average 10.8% higher than white wavelength. Similarly, lipid productivity was highest in blue wavelength, at least 46.8% higher than white wavelength, whereas FAME composition revealed a mild increase in oleic and palmitic acid levels. Cost analysis reveals that raceways treating wastewater and using monochromatic wavelength would decrease costs from 2.71 to 0.73 $/kg biomass. We prove that increasing both biomass and lipid productivity is possible through cost-effective approaches, thereby accelerating the commercialization of low-value products from microalgae, like biodiesel.

• Microbiology and Biotechnology Letters
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• v.47 no.3
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• pp.381-389
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• 2019

Chlorella vulgaris was isolated from the Nile River, Qalubia Governorate, Egypt, for possible use in biodiesel production. BG-II nutrient growth media was used for isolation and laboratory growth. Identification was performed via 18S rRNA gene amplification, followed by sequencing. The alga was exposed to UV-C (254 nm) for 15, 30, and 45 s to improve dry weight accumulation and to increase the oil production. Daily measurements of dry weight ($g{\cdot}l^{-1}$) were performed; oil content and volumetric lipid productivity were also determined. UV-C exposure led to an increase in the volumetric lipid productivity by 27, 27.3, and $32.4mg{\cdot}l^{-1}{\cdot}d^{-1}$ with 15, 30, and 45 s, respectively, as compared with the control, which resulted in $18mg{\cdot}l^{-1}{\cdot}d^{-1}$. Of the examined mutants, the one with the highest productivity was re-irradiated by UV-C (254 nm) for 15, 30, 45, and 60 s. For 15 s of exposure time, the oil content increased to 34%, while it was 31% at 30 s; further, it decreased to 22% at 45 and 60 s exposures. The fatty acid methyl ester profile was 82.22% in the first mutant at 45 s, compared with the wild strain that contained a total of 66.01% of FAs. Furthermore, the highest levels of polyunsaturated fatty acid methyl ester were observed in the mutant exposed for 45 s, and it reached 11.41%, which reduced the cetane number to 71.3.

• Journal of Microbiology and Biotechnology
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• v.32 no.11
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• pp.1357-1372
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• 2022

Heavy reliance on fossil fuels has been associated with increased climate disasters. As an alternative, microalgae have been proposed as an effective agent for biomass production. Several advantages of microalgae include faster growth, usage of non-arable land, recovery of nutrients from wastewater, efficient CO₂ capture, and high amount of biomolecules that are valuable for humans. Microalgae Chlorella spp. are a large group of eukaryotic, photosynthetic, unicellular microorganisms with high adaptability to environmental variations. Over the past decades, Chlorella has been used for the large-scale production of biomass. In addition, Chlorella has been actively used in various food industries for improving human health because of its antioxidant, antidiabetic, and immunomodulatory functions. However, the major restrictions in microalgal biofuel technology are the cost-consuming cultivation, processing, and lipid extraction processes. Therefore, various trials have been performed to enhance the biomass productivity and the lipid contents of Chlorella cells. This study provides a comprehensive review of lipid enhancement strategies mainly published in the last five years and aimed at regulating carbon sources, nutrients, stresses, and expression of exogenous genes to improve biomass production and lipid synthesis.