• Journal of Microbiology and Biotechnology
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• v.26 no.5
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• pp.854-866
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• 2016

The production cost of biodiesel from microalgae is still not competitive, compared with that of petroleum fuels. The genetic improvement of microalgal strains to increase triacylglycerol (TAG) accumulation is one way to reduce production costs. One of the most promising approaches is the isolation of starch-deficient mutants, which have been reported to successfully increase TAG yields. To date, such a stable mutant is not available in an oleaginous marine microalga, despite several advantages of using marine species for biodiesel production. Algae in the genus Dunaliella are known to tolerate high salt concentration and other environmental stresses. In addition, the cultivation processes for large-scale outdoor commercialization have been well established for this genus. In this study, Dunaliella tertiolecta was used to screen for starch-deficient mutants, using an iodine vapor-staining method. Four out of 20,016 UV-mutagenized strains showed a substantial reduction of starch content. A significantly higher TAG content, up to 3-fold of the wild-type level, was observed in three of the mutants upon induction by nitrogen depletion. The carotenoid production and growth characteristics of these mutants, under both normal and oxidative stress conditions, were not compromised, suggesting that these processes are not necessarily affected by starch deficiency. The results from this work open up new possibilities for exploring Dunaliella for biodiesel production.

• 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.30 no.10
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• pp.1597-1606
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• 2020

Transcription factor engineering to regulate multiple genes has shown promise in the field of microalgae genetic engineering. Here, we report the first use of transcription factor engineering in Chlorella sp. HS2, thought to have potential for producing biofuels and bioproducts. We identified seven endogenous bZIP transcription factors in Chlorella sp. HS2 and named them HSbZIP1 through HSbZIP7. We overexpressed HSbZIP1, a C-type bZIP transcription factor, in Chlorella sp. HS2 with the goal of enhancing lipid production. Phenotype screening under heterotrophic conditions showed that all transformants exhibited increased fatty acid production. In particular, HSbZIP1 37 and 58 showed fatty acid methyl ester (FAME) yields of 859 and 1,052 mg/l, respectively, at day 10 of growth under heterotrophic conditions, and these yields were 74% and 113% higher, respectively, than that of WT. To elucidate the mechanism underlying the improved phenotypes, we identified candidate HSbZIP1-regulated genes via transcription factor binding site analysis. We then selected three genes involved in fatty acid synthesis and investigated mRNA expression levels of the genes by qRT-PCR. The result revealed that the possible HSbZIP1-regulated genes involved in fatty acid synthesis were upregulated in the HSbZIP1 transformants. Taken together, our results demonstrate that HSbZIP1 can be utilized to improve lipid production in Chlorella sp. HS2 under heterotrophic conditions.

• Journal of the Korean Society of Fisheries and Ocean Technology
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• v.51 no.4
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• pp.475-483
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• 2015

The marine microalgae Isochrysis galbana was cultured under various light-emitting diode (LED) light conditions with different wavelengths to examine changes in growth and in amino acid and fatty acid profiles. The culture conditions for the microalgae were Conway medium, salinity of 33 psu, temperature of $24^{\circ}C$, and a 16/8 h light/dark photoperiod. Six light sources, including 5 units of 180W LED lamps (peak wavelength: blue [LB] 470 nm; green [LG] 525 nm; yellow [LY] 595 nm; red [LR] 636 nm; white [LW] 442 nm) and 1 unit of a 175W metal halide (MH) lamp, were used for the experiment. The dry cell weights ($gL^{-1}$) of I. galbana under different light conditions were in the order of LW>LB${\geq}$MH>LR>LG>LY. Levels of essential amino acids were revealed to be significantly higher under LW, LG, and MH than under the other wavelengths (P<0.05). The fatty acid, unsaturated fatty acid, and DHA contents of I. galbana were higher under MH, LW, and LG. In addition, the carotenoid content was higher under MH, LW, and LG than under the other wavelengths (P<0.05). The fucoxanthin content was highest under MH (0.28%) and lowest under LY (0.2%), and it was 0.26% under LW and LG. The results indicate that the combined use of LW and LG is effective when using LED lamps for I. galbana cultivation.

• Journal of the Korea Organic Resources Recycling Association
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• v.9 no.1
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• pp.73-78
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• 2001

The biological carbon dioxide fixation using microalgae and photo-bioreactor has been known as an effective carbon dioxide reduction technology. As algae has many other environmental factor for its growth, the desirable cultivation factors were investigated using a green alga, Euglena gracilis Z. In this study, Euglena gracilis Z showed good $CO_2$ fixation ability in high $CO_2$ concentration of 10-20% and it contained the high protein and vitamin E enough to be used as fodder. For the mass cultivation, the continuous and semi-continuous cultivation methods were employed. The optimum hydraulic retention time (HRT) for the continuous cultivation was 4 days at carbon dioxide concentration of 10%. In this condition, the final cell number was $3.57{\times}10^6/m{\ell}$. The growth of Euglena gracilis Z increased according to the light intensity.

• Applied Chemistry for Engineering
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• v.19 no.2
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• pp.145-150
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• 2008

In this work we have studied the antifouling properties of the hydrophobic sol-gel modified sensing membrane and its optical properties for sensor application. E. coli JM109, B. cereus 318 and P. pastoris X-33 were cultivated in confocal cultivation dishes with glass surface, respectively. The glass surface was coated with the hydrophobic sol-gels prepared by the dimethoxy-dimethyl-silane (DiMe-DMOS) and tetramethyl-orthosilicate (TMOS). After cultivation, microorganisms adhered on the surface coated with sol-gels and glass surface were dyed by gram-staining method and the numbers of microorganisms were analyzed based on the image data of the scanning electronic microscope (SEM). A great number of microorganisms, about $2{\sim}3{\times}10^4/mm^2$, was adhered on the glass surfaces which no hydrophobic sol-gels were coated. But, the antifouling effect of the hydrophobic sol-gels was large, that microorganisms of less than $200{\sim}300/mm^2$ were adhered on the coated glass surface. The performance of the sensing membranes for detection of pH and dissolved oxygen was enhanced by recoating the light insulation layer prepared with the mixture of the hydrophobic sol-gel and graphite particles.

• Journal of Marine Bioscience and Biotechnology
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• v.15 no.2
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• pp.82-89
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• 2023

Microalgae, as photosynthetic organisms, possess the ability to produce a diverse array of bioactive compounds. This study focused on the transformant Chlamydomonas reinhardtii dZL and subjected it to cultivation under varying light intensities (60, 120, 180, and 240 µmol/m²/s). Our aim was to assess the impact of light intensity on both microalgal biomass and carotenoid production. The cultivation took place in 80 mL bubble column photobioreactors, specifically the Multi-cultivator. Notably, the culture exposed to 240 µmol/m²/s exhibited the most rapid cell growth, surpassing even the cell concentration achieved at 180 µmol/m²/s by day 8. A detailed analysis of the specific irradiance rate over time unequivocally revealed a sharp decline in growth rates when the rate fell below 2 × 10^-10 µmol/cell/s. Although the culture with 60 µmol/m²/s yielded the highest carotenoid content (1.2% of dry weight), the culture exposed to 240 µmol/m²/s recorded the highest carotenoid concentration at 8.9 mg/L owing to its higher biomass. Our findings reveal the critical importance of maintaining a specific irradiance rate above 2 × 10^-10 µmol/cell/s to enhance biomass and carotenoid productivity. This study lays the groundwork for defining optimal light intensity conditions applicable to mass culture systems, with the objective of augmenting C. reinhardtii biomass and optimizing carotenoid productivity.

• Journal of Plant Biotechnology
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• v.45 no.4
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• pp.400-408
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• 2018

Biodiversity has continued to degrade in the $21^{st}$ century due to global warming occasioned by destruction of the environment around the world.. The Nagoya protocol places Korea in a unique position to effectively develop and protect its domestic genetic resources. Microalgae under study in this research contains large amount of antioxidant substances such as beta carotene and astaxanthin, that can be used as biological resource owing to the large amounts of biomass that can be secured through photosynthesis. However, it is difficult to preserve it since cryopreservation method used for long-term preservation is yet to be developed. A basic study for long term cryopreservation was carried out on Nizschia frustulum and Nitzschia amabilis which belong to marine diatoms. As cryoprotectants (CPAs), glycerol, DMSO, and methanol which penetrate into cells were prepared at 5%, 10%, and 15% concentrations each, in case of methanol, it was tested at concentrations of 5%, 10% and 12% by its nature. Two kinds of microalgae, N. frustulum and N. amabilis, were diluted with $10^2$, $10^3$ and $10^4cells\;ml^{-1}$, respectively. The highest survival rate was shown at12% concentration of methanol, and the figures were $6.94{\pm}0.31%$ in N. frustulum and $8.85{\pm}0.16%$ in N. amabilis. As a result of 3 weeks cultivation of thawed microalgae after freezing, the result is shows that N. frustulum increased about 10 times faster and N. amabilis increased about 12 times the original concentration.

• Korean Journal of Food Science and Technology
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• v.47 no.6
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• pp.687-697
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• 2015

With the shortage of edible fats and oils and depletion of fossil fuels in many countries, microbial lipids is emerging as one of the most promising sources of fats and oils in the global market. Oleaginous microorganisms, also called single cell oils (SCOs), can accumulate lipids more than 25% in the cell volume. Triacylglycerols are the major storage lipids. SCOs offer several advantages for lipid production as follows: SCOs have short life span which would shorten production time, cultivation conditions are not affected by climate and place; the production process is easy to control. There are a number of oleaginous yeasts, molds, and microalgae. Furthermore, the lipid productivity of SCOs can be enhanced through strain improvement and the optimization of cultivation conditions. The new strains developed using recent advanced biotechnical methods showed greatly improved oleaginicity. Further, hydrolysates of lignocellulosic materials can be used as carbon sources for economic production of SCO.

• Environmental Engineering Research
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• v.25 no.3
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• pp.393-399
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• 2020

The use of different types of wastewater (WW) for the cultivation of microalgae and cyanobacteria during recent decades has provided important economic and environmental benefits. However, direct use of WW can lead to growth inhibition and biomass contamination. In the present study, we separated the key WW nutrients, namely nitrate and phosphate, by adsorption and regeneration and used the resulting regenerated water to cultivate the cyanobacterium Spirulina platensis. The adsorbent was granular γ-alumina derived from waste aluminum cans. This procedure recovered 19.9% of nitrate and 23.7% of phosphate from WW. The cyanobacterial cultures efficiently assimilated the nutrients from the medium prepared using regenerated WW, and the growth and nutrient uptake were similar to those in a synthetic medium. In addition, imposing nutrient limitations to increase carbohydrate productivity was easily achieved using regenerated wastewater nutrients, without requiring additional dilution or complex processing. In acute toxicity tests, the harvested biomass in a regenerated medium had similar toxicity levels compared to the biomass obtained from a synthetic medium. The proposed method of using regenerated WW to produce contamination-free biomass has broad potential applications.