• Environmental Engineering Research
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• v.20 no.4
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• pp.377-382
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• 2015

The high concentration of nitrogen and phosphorus in wastewater incorporated with the ability to use carbon dioxide as the carbon source make the microalgae become more attractive in wastewater treatment process. This study evaluates the optimal conditions for the digestion of settelable solids from the recirculating aquaculture system to produce the biomass of the green microalga Scenedesmus sp. After solids separation, aerobic digestion of settleable solids under disperse condition produced nitrate as the final product of consequently ammonification and nitrification processes. With the optimal digestion procedure, nitrate concentration during aerobic digestion in 2000 mL vessel increased from $9.63{\pm}0.65mg\;N/L$ to $58.66{\pm}0.06mg\;N/L$ in 10 days. Thereafter, cultivation of Scenedesmus sp. was performed in 1000 mL Duran bottle with air bubbling. The highest Scenedesmus sp. specific growth rate of $0.321{\pm}0.01/d$ was obtained in treatment using liquid fraction after aerobic digestion as the whole culture medium for Scenedesmus sp. cultivation. With this study, digestion of $8,800{\pm}128.12mg\;dry\;weight/L$ of settleable solids from fish pond finally produced $1,235{\pm}21mg\;dry\;weight/L$ of Scenedesmus sp. biomass.

• Journal of Microbiology and Biotechnology
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• v.26 no.3
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• pp.503-510
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• 2016

The accumulation (internal and superficial distribution) of magnesium ions (Mg²⁺) by the green freshwater microalga Chlorella vulgaris (C. vulgaris) was investigated under autotrophic culture in a stirred photobioreactor. The concentrations of the three forms of Mg²⁺ (dissolved, extracellular, and intracellular) were determined with atomic absorption spectroscopy during the course of C. vulgaris growth. The proportions of adsorbed (extracellular) and absorbed (intracellular) Mg²⁺ were quantified. The concentration of the most important pigment in algal cells, chlorophyll a, increased over time in proportion to the increase in the biomass concentration, indicating a constant chlorophyll/biomass ratio during the linear growth phase. The mean-average rate of Mg²⁺ uptake by C. vulgaris grown in a culture medium starting with 16 mg/l of Mg²⁺ concentration was measured. A clear relationship between the biomass concentration and the proportion of the Mg²⁺ removal from the medium was observed. Of the total Mg²⁺ present in the culture medium, 18% was adsorbed on the cell wall and 51% was absorbed by the biomass by the end of the experiment (765 h). Overall, 69% of the initial Mg²⁺ were found to be removed from the medium. This study supported the kinetic model based on a reversible first-order reaction for Mg²⁺ bioaccumulation in C. vulgaris, which was consistent with the experimental data.

• ALGAE
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• v.31 no.3
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• pp.267-276
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• 2016

Microalgae are currently a very promising source of biomass and triacylglycerol (TAG) for biofuels. In a previous study, we identified Chlorella saccharophila as a suitable source of oil for biodiesel production because it showed high biomass and lipid content with an appropriate fatty acid methyl esters profile. To improve the TAG accumulation in C. saccharophila, in this study we evaluated the effect of abscisic acid (ABA) addition on cell concentration, lipid content and TAG production in this microalga. First, we evaluated the effects of four ABA concentrations (1, 4, 10, and 20 μM) added at the beginning of a single-stage cultivation strategy, and found that all concentrations tested significantly increased cell concentration and TAG content in C. saccharophila. We then evaluated the addition of 1 μM ABA during the second stage of a two-stage cultivation strategy and compared it with a nitrogen deficiency treatment (ND) and a combination of ND and ABA (ND + ABA). Although ABA alone significantly increased lipid and TAG contents compared with the control, ND showed significantly higher TAG content, and ND + ABA showed the highest TAG content. When comparing the results of both strategies, we found a superior response in terms of TAG accumulation with the addition of 1 μM ABA at the beginning of a single-stage cultivation system. This strategy is a simple and effective way to improve the TAG content in C. saccharophila and probably other microalgae as a feedstock for biodiesel production.

• ALGAE
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• v.37 no.2
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• pp.175-183
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• 2022

Polar microorganisms produce physiologically active substances to adapt to harsh environments, and these substances can be used as biomedical compounds. The green microalga Micractinium variabile KSF0031, which was isolated from Antarctica, produced phytol, a natural antimicrobial agent. Furthermore, several polyunsaturated fatty acids (PUFAs), including omega-3, exhibit antioxidant properties. Here statistical methods (Plackett-Burman design and Box-Behnken design) were used to optimize the culture medium of KSF0031 to improve biomass production, and K₂HPO₄, MgSO₄·7H ₂O, and ammonium ferric citrate green (AFCg) were selected as significant components of the culture medium. Changes in the concentration of K₂HPO₄ and MgSO₄·7H ₂O as positive factors and AFCg as a negative factor affected cell growth to a remarkable degree. The biomass production in a 100 L culture using the optimized medium for 24 d at 18℃ was improved by 37.5% compared to that obtained using the original BG-11 medium. The quantities of PUFAs and phytol obtained were 13 mg g^-1 dry cell weight (DCW) and 10.98 mg g^-1 DCW, which represent improved yields of 11.70% and 48.78%, respectively. The results of this study could contribute to an improved production of phytol and fatty acids from Antarctic microalgae in the biomedical industry.

• Journal of Microbiology and Biotechnology
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• v.15 no.6
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• pp.1159-1163
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• 2005

The production of hydrogen by Chlamydomonas reinhardtii UTEX 90, a marine green alga, was performed under dark fermentation. The effects of initial nitrogen and phosphorus concentration on the cell growth and the production of hydrogen and organic substances were investigated. In the growth stage, the maximum dry cell weight (DCW) was 3 g/l when the initial ammonium concentration was 15 mM. In the dark fermentation, the maximum hydrogen production was $3.5\;{\mu}mol/\;mg$ DCW when the initial nitrogen concentration was 7.5 mM. The nitrogen concentration had a greater effect on organic compound and hydrogen production than the phosphorus concentration during the dark fermentation. An investigation of the duration of dark fermentation showed that, at least until three days, dark fermentation should be prolonged for maximum hydrogen production.

• ALGAE
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• v.28 no.4
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• pp.379-387
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• 2013

The purpose of this study was to express bovine lactoferrin N-lobe in Chlorella vulgaris, a green microalga, using the pCAMBIA1304 vector. Chlorella-codon-optimized bovine lactoferrin N-lobe (Lfb-N gene) was cloned in the expression vector pCAMBIA1304, creating the plasmid pCAMLfb-N. pCAMLfb-N was then introduced into C. vulgaris by electro-transformation. Transformants were separated from BG-11 plates containing 20 ${\mu}g\;mL^{-1}$ hygromycin. Polymerase chain reaction was used to screen transformants harboring Lfb-N gene. Finally, total soluble protein was extracted from the transformants, and the expression of Lfb-N protein was detected using western blotting. Using this method, we successfully expressed bovine lactoferrin in C. vulgaris. Therefore, our results suggested that recombinant lactoferrin N-lobe, which has many uses in the biomedical and pharmaceutical industries, can be produced economically.

• Korean Chemical Engineering Research
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• v.44 no.1
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• pp.46-51
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• 2006

As one of the $CO_2$ reduction strategies, a biological method was proposed to convert $CO_2$ to useful biomass with antioxidant carotenoids by photosynthetic microorganisms. One of the photoautotrophs, Haematococcus pluvialis is a freshwater green microalga and accumulates the secondary carotenoid astaxanthin during induction of green vegetative cells to red cyst cells. In this study, $CO_2$ fixation and astaxanthin production using H. pluvialis was conducted by photoautotrophic culture in the $CO_2$ supplemented photo-incubator. Maximum growth rate of H. pluvialis was obtained at a 5％ $CO_2$ environment on basic N and P conditions of NIES-C medium. The photoautotrophic induction consisted of 5％ $CO_2$ supply and high light illumination promoted astaxanthin synthesis in H. pluvialis, yielding an astaxanthin productivity of $9.6mg/L{\cdot}day$ and a $CO_2$ conversion rate of $27.8mg/L{\cdot}day$ to astaxanthin. From the results the sequential photoautotrophic culture and induction process using H. pluvialis is expecting an alternative $CO_2$ reduction technology with a function of valuable biosubstance production.

• Journal of Microbiology and Biotechnology
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• v.17 no.3
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• pp.414-419
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• 2007

Botryococcus braunii is a green colonial fresh water microalga and it is recognized as one of the renewable resources for production of liquid hydrocarbons. CFTRI-Bb-l and CFTRI-Bb-2 have been reported for the first time and their performance with regard to growth and biochemical profile is presented here. The present study focused on effect of carbon dioxide $(CO_2)$ on biomass, hydrocarbon, carbohydrate production, fatty acid profile, and carotenoid content in various species of B. braunii (LB-572, SAG 30.81, MCRC-Bb, N-836, CFTRI-Bb-l, and CFTRI-Bb-2) at 0.5, 1.0, and 2.0% (v/v) levels using a two-tier flask. $CO_2$ at 2.0% (v/v) level enhanced growth of the organism, and a two-fold increase in biomass and carotenoid contents was observed in all the B. braunii strains studied compared with control culture (without $CO_2$ supplementation). At 1 % and 2% (v/v) $CO_2$ concentrations, palmitic acid and oleic acid levels increased by 2.5 to 3 folds in one of the strains of B. braunii (LB-572). Hydrocarbon content was found to be above 20% at 2% $CO_2$ level in the B. braunii LB-572, CFTRI-Bb-2, CFTRI-Bb-l, and N-836 strains, whereas it was less than 20% in the SAG 30.81 and MCRC-Bb strains compared with control culture. This culture methodology will provide information on $CO_2$ requirement for growth of algae and metabolite production. B. braunii spp. can be grown at the tested levels of $CO_2$ concentration without much influence on culture pH.

• Environmental Engineering Research
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• v.25 no.2
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• pp.135-146
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• 2020

This study was aimed to investigate the possibility of using raw and anaerobically-digested piggery wastewater as culture media for a green microalga Chlorella vulgaris (C. vulgaris). Due to high concentration of ammonia and dark color, the microalga did not grow well in this wastewater. In order to solve this problem, air stripping and NaOCl-treatment were applied to reduce the concentration of NH₃-N and the color intensity from the wastewater. Algal growth was monitored in terms of specific growth rate, biomass productivity, and nutrient removal efficiency. As a result, C. vulgaris grew without any sign of inhibition in air-stripped and 10-folds diluted anaerobically-digested piggery wastewater with enhanced biomass productivity of 0.57 g/L·d and nutrient removal of 98.7-99.8% for NH₃-N and 41.0-62.5% for total phosphorus. However, NaOCl-treatment showed no significant effect on growth of C. vulgaris, although dark color was removed greatly. Interestingly, despite that the soluble organic concentration after air stripping was still high, the biomass productivity was 4.4 times higher than BG-11. Moreover, air stripping was identically effective for raw piggery wastewater as for anaerobic digestate. Therefore, it was concluded that air stripping was a very effective method for culturing microalgae and removing nutrients from raw and anaerobically-digested piggery wastewaters.

• Journal of Microbiology and Biotechnology
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• v.21 no.10
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• pp.1073-1080
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• 2011

In order to investigate and generalize the effects of carbon and nitrogen sources on the growth of and lipid production in Chlorella sp. 227, several nutritional combinations consisting of different carbon and nitrogen sources and concentrations were given to the media for cultivation of Chlorella sp. 227, respectively. The growth rate and lipid content were affected largely by concentration rather than by sources. The maximum specific growth was negatively affected by low concentrations of carbon and nitrogen. There is a maximum allowable inorganic carbon concentration (less than 500~1,000 mM bicarbonate) in autotrophic culture, but the maximum lipid content per gram dry cell weight (g DCW) was little affected by the concentration of inorganic carbon within the concentration. The lipid content per g DCW was increased when the microalga was cultured with the addition of glucose and bicarbonate (mixotrophic) at a fixed nitrogen concentration and with the lowest nitrogen concentration (0.2 mM), relatively. Considering that lipid contents per g DCW increased in those conditions, it suggests that a high ratio of carbon to nitrogen in culture media promotes lipid accumulation in the cells. Interestingly, a significant increase of the oleic acid amount to total fatty acids was observed in those conditions. These results showed the possibility to induce lipid production of high quality and content per g DCW by modifying the cultivation conditions.