• Journal of Microbiology and Biotechnology
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• v.32 no.3
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• pp.378-386
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• 2022

Scenedesmus obliquus ABC-009 is a microalgal strain that accumulates large amounts of lutein, particularly when subjected to growth-limiting conditions. Here, the performance of this strain was evaluated for the simultaneous production of lutein and biofuels under three different modes of cultivation - photoautotrophic mode using BG-11 medium with air or 2% CO₂ and heterotrophic mode using YM medium. While it was found that the highest fatty acid methyl ester (FAME) level and lutein content per biomass (%) were achieved in BG-11 medium with CO₂ and air, respectively, heterotrophic cultivation resulted in much higher biomass productivity. While the cell concentrations of the cultures grown under BG-11 and CO₂ were largely similar to those grown in YM medium, the disparity in the biomass yield was largely attributed to the larger cell volume in heterotrophically cultivated cells. Post-cultivation light treatment was found to further enhance the biomass productivity in all three cases and lutein content in heterotrophic conditions. Consequently, the maximum biomass (757.14 ± 20.20 mg/l/d), FAME (92.78 ± 0.08 mg/l/d), and lutein (1.006 ± 0.23 mg/l/d) productivities were obtained under heterotrophic cultivation. Next, large-scale lutein production using microalgae was demonstrated using a 1-ton open raceway pond cultivation system and a low-cost fertilizer (Eco-Sol). The overall biomass yields were similar in both media, while slightly higher lutein content was obtained using the fertilizer owing to the higher nitrogen content.

• KSBB Journal
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• v.24 no.4
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• pp.377-382
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• 2009

The culture condition of growing Chlorella minutissima was optimized to produce biodiesel for fed-batch cultivation. First, under heterotrophic cultivation, the optimum level of glucose was determined to be 10 g/L for 20 days. After, three cultivation conditions were operated: autotrophic, heterotrophic, and mixotrophic growth. The lipid level and the maximum cell concentration from the fed-batch heterotrophic process were 32.0 (%, v/v) and 15.0 (g-dry wt./L) in 20 L flask, respectively. In addition, since the relatively constant specific lipid production rate was observed as 0.040 (% lipid/g-dry wt./day) at the latter period of cultivation time, the fed-batch process could maintain continuous lipid production. Fed-batch process is higher than those values from the batch process. The lipids from the fed-batch process contained over 38% of $C_{18}$, known as the suitable composition for the biodiesel application. For mixotrophic and heterotrophic growth under fed-batch condition, glucose was proved to be an appropriate carbon source for a large scale outdoor cultivation. For fed-batch cultivation, the feeding rate of seawater medium containing glucose was decided to be 0.5 L/day. The mixotrophic cultivation maintained maximum cell concentration of 24 (g-dry wt./L) and the lipid level of 43 (%, w/w). The lipid composition from this process was also proved to be suitable for the biodiesel production. The fatty acids from the mixotrophic growth contains 18% of $C_{17}$ and 49% of $C_{18}$, implying It also tells that C. minutissima is a suitable resource of biodiesel. Especially, the mixotrophic cultivation with fed-batch process might be useful for the large scale cultivation for the biodiesel production.

• Applied Chemistry for Engineering
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• v.33 no.2
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• pp.222-228
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• 2022

As a possible feedstock for biodiesel, the marine green alga Tetraselmis sp. was cultivated under different conditions of phototrophic, mixotrophic and heterotrophic cultures. Glycerol, a byproduct from biodiesel production process, was used as the carbon source of mixotrophic and heterotrophic culture. The effects of glycerol supply and nitrate-repletion were compared for different trophic conditions. Mixotrophic cultivation exhibited higher biomass productivity than that of phototrophic and heterotrophic cultivation. Maximum lipid productivity of 55.5 mg L^-1 d^-1 was obtained in the mixotrophic culture with 5 g L^-1 of glycerol and 8.8 mM of nitrate due to the enhancement of both biomass and lipid accumulation. The major fatty acid methyl esters (FAME) in the produced biodiesel were palmitic acid (C16:0), oleic acid (C18:1), linoleic acid (C18:2), and linolenic acid (C18:3). The degree of unsaturation was affected by different culture conditions. The biodiesel properties predicted by correlation equations based on the FAME profiles mostly complied with the specifications from the US, Europe and Korea, with the exception of the cold-filter plugging point (CFPP) criterion of Korea.

• Microbiology and Biotechnology Letters
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• v.35 no.1
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• pp.45-51
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• 2007

Arthrospira (Spirulina) platensis is one of the commercially important filamentous cyanobacteria. The heterotrophic cultivation of Arthrospira can be an alternative strategy for commercial mass production. In heterotrophic culture, the specific growth rate of A. platensis M9108, a glucose-resistant mutant of A. platensis PCC 9108, was $0.014h^{-1}$ which was 1.8 higher than that oi the previous report. The mutant possessed the facility to assimilate and to metabolize glucose efficiently under heterotrophic condition. However, the r-linolenic acid content of 6 Arthrospira strains was not increased in heterotrophic culture. Four Arthrospira strains out of 6 tested strains were able to utilize maltose as a carbon source under heterotrophic condition. The biomass production of these strains on maltose was similar to that on glucose. The specific growth rate of A. platensis M9108 increased with glucose concentration up to 5.0 g/L and then decreased at a glucose concentration of 10.0 g/L. Additionally, A. platensis M9108 under heterotrophic condition showed no aggregation during the cultivation in contrast to A. platensis PCC 9108.

• Journal of Korean Society of Water and Wastewater
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• v.27 no.6
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• pp.703-709
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• 2013

Among many microalgae cultivation types, heterotrophic culture with low cost carbon sources and energy saving culture method is crucial. A result of estimating the effects of pH on wastewater treatment using heterotrophic growing microalgae Chlorella sorokiniana shows that there was no difference in microalgae growth amount and nitrogen, phosphorus removal rate by wide range of pH(5 ~ 9). From pH 5 to 9, total nitrogen, phosphorous and glucose removal rates were 10.5 mg-N/L/d, 2 mg-P/L/d, 800 ~ 1000 mg/L respectively. This study reveals that C. sorokiniana cannot metabolite glycerol heterotrophically, however, glucose and acetate were proper carbon sources for growth and T-N, T-P and TOC removal. This research highlights the potential of heterotrophic microalgal growth with wastewater treatment plant with wide range of pH and carbon sources.

• Journal of Marine Bioscience and Biotechnology
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• v.6 no.2
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• pp.68-75
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• 2014

This study investigated the potential use of various organic carbon sources (glucose, glycerol and acetate) and different concentrations of $CO_2$ for culturing marine microalga Dunaliella tertiolecta. Cell growth and lipid production were monitored under heterotrophic, mixotrophic and photoautotrophic modes of cultivation. D. tertiolecta showed the ability to grow under mixotrophic (acetate and glucose), heterotrophic (glucose) and photoautotrophic condition under high $CO_2$ concentration (15%). With all the organic carbon sources (glucose, glycerol and acetate) tested in this study, 1~5% acetate enhanced cell growth rate and lipid content, while higher concentrations of acetate (10% and 15%) were inhibitory and resulted in cell death.

• 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.

• ALGAE
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• v.28 no.2
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• pp.131-147
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• 2013

Major progress has been made in the past decade towards understanding of the biosynthesis of red carotenoid astaxanthin and its roles in stress response while exploiting microalgae-based astaxanthin as a potent antioxidant for human health and as a coloring agent for aquaculture applications. In this review, astaxanthin-producing green microalgae are briefly summarized with Haematococcus pluvialis and Chlorella zofingiensis recognized to be the most popular astaxanthin-producers. Two distinct pathways for astaxanthin synthesis along with associated cellular, physiological, and biochemical changes are elucidated using H. pluvialis and C. zofingiensis as the model systems. Interactions between astaxanthin biosynthesis and photosynthesis, fatty acid biosynthesis and enzymatic defense systems are described in the context of multiple lines of defense mechanisms working in concert against photooxidative stress. Major pros and cons of mass cultivation of H. pluvialis and C. zofingiensis in phototrophic, heterotrophic, and mixotrophic culture modes are analyzed. Recent progress in genetic engineering of plants and microalgae for astaxanthin production is presented. Future advancement in microalgal astaxanthin research will depend largely on genome sequencing of H. pluvialis and C. zofingiensis and genetic toolbox development. Continuous effort along the heterotrophic-phototrophic culture mode could lead to major expansion of the microalgal astaxanthin industry.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.49 no.1
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• pp.38-44
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• 2016

The quality and quantity of live food sources strongly influence the success of fish production in farming operations. Thus, critical studies of live forage species are a crucial element for progress in fish aquaculture. The fat content of food is an especially important determinant of growth in marine fish. Omega-3 highly unsaturated fatty acids (HUFA) are essential components of diet that determine the nutritional value of larval fish. Euglena is a protist that has potential as a forage species. These single-celled organisms have plant and animal characteristics they are motile, elliptical in shape and 15–500 μm in diameter. Their nutritional content is excellent, but most studies have focused on cells raised in autotrophic culture. We therefore examined differences in the lipid and fatty acid contents, and the growth of Euglena cells grown under autotrophic, heterotrophic, and mixotrophic conditions. Biomass production reached 15.03 g/L, 12.28 g/L, and 3.66 g/L under mixotrophy, heterotrophy, and autotrophy, respectively. The proportional n-3 HUFA content differed among culture methods: 10.04%, 5.80% and 10.01% in mixotrophic, heterotrophic and autotrophic cultures, respectively. Mixotrophy was to be the best form of cultivation for improving the growth and nutritional content of Euglena.

• Applied Chemistry for Engineering
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• v.32 no.5
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• pp.487-496
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• 2021

Microalgae is one of the promising biodiesel feedstock with high growth rates compared to those of terrestrial oil crops. Despite its numerous advantages, biodiesel production from microalgae needs to reduce energy demand and material costs further to go to commercialization. During solvent extraction of microalgal lipids, lipid-extracted algae (LEA) cell residue is generated as an organic solid waste, about 80-85% of original algal biomass, and requires an appropriate recycling or economic disposal. The resulting LEA still contains significant amount of carbohydrates, proteins, N, P, and other micronutrients. This review will focus on recent advancement in the utilization of LEA as: (i) utilization as nutrients or carbon sources for microalgae and other organisms, (ii) anaerobic digestion to produce biogas or co-fermentation to produce CH₄ and H₂, and (iii) conversion to other forms of biofuel through thermochemical degradation processes. Possible mutual benefits in the integration of microalgae cultivation-biodiesel production-resulting LEA with anaerobic digestion and thermochemical conversion are also discussed.