Myounghoon Moon

Rapid quantification of microalgal lipids in aqueous medium by a simple colorimetric method.

Identification of novel microalgal strains with high lipid productivity is one of the most important research topics in renewable biofuel research. However, the major bottleneck in the strain screening process is that currently known methods for the estimation of microalgal lipid are laborious and time-consuming. The present study successfully employed sulpho-phospho-vanillin (SPV) colorimetric method for direct quantitative measurement of lipids within liquid microalgal culture. The SPV reacts with lipids to produce a distinct pink color, and its intensity can be quantified using spectrophotometric methods by measuring absorbance at 530nm. This method was employed for a rapid quantification of intracellular lipid contents within Chlorella sp., Monoraphidium sp., Ettlia sp. and Nannochloropsis sp., all of which were found to have lipid contents ranging in between 10% and 30%. Subsequent analysis of the biomass using gas chromatography confirmed that our protocol is highly accurate (R(2)=0.99).

Effect of monochromatic illumination on lipid accumulation of Nannochloropsis gaditana under continuous cultivation.

Although nitrogen starvation is frequently used to increase lipid contents in microalgae, it has a negative effect on cellular growth. Since light supply is essential for photosynthetic organisms, the effects of cultivation under monochromatic illumination on the growth and lipid contents of Nannochloropsis gaditana were assessed. Continuous cultivation under blue and red light conditions improved the productivity and physical properties for biodiesel from this microalga. FAME yield was twofold higher under red light than under normal white light (21.12% vs 11.35%), with no significant difference in growth rates. Blue and red light increased photosynthetic oxygen evolution, carbon fixation and nutrient uptake. In total, more significant physiological changes were observed under red than under blue light. These results show that red light illumination may be useful for enhancing lipid production by N. gaditana, with the increased photosynthetic reducing equivalents induced by red light which could be deposited as lipids and carbohydrates.

Enhancing lipid productivity of Chlorella vulgaris using oxidative stress by TiO2 nanoparticles

Ability to increase the lipid production in microalgae is one of the heavily sought-after ideas to improve the economic feasibility of microalgae-derived transportation fuels for commercial applications. We used the oxidative stress by TiO2 nanoparticles, a well-known photocatalyst, to induce lipid production in microalgae. Chlorella vulgaris UTEX 265 was cultivated under various concentrations of TiO2 ranging from 0.1 to 5 g/L under UV-A illumination. Maximum specific growth rate was affected in responding to TiO2 concentrations. In the presence of UV-A, chlorophyll concentration was decreased at the highest concentration of TiO2 (5 g/L TiO2) by oxidative stress. The fatty acid ethyl ester (FAME) composition analysis suggested that oxidative stress causes the accumulation and decomposition of lipids. The highest FAME productivity was 18.2 g/L/d under low concentrations of TiO2 (0.1 g/L) and a short induction time (two days). The controlled condition of TiO2/UV-A inducing oxidative stress (0.1 g/L TiO2 and two days induction) could be used to increase the lipid productivity of C. vulgaris UTEX 265. Our results show the possibility of modulating the lipid induction process through oxidative stress with TiO2/UV-A.

Development of direct conversion method for microalgal biodiesel production using wet biomass of Nannochloropsis salina.

In this work, the effects of several factors, such as temperature, reaction time, and solvent and acid quantity on in situ transesterification yield of wet Nannochloropsis salina were investigated. Under equivalent total solvent volume to biomass ratio, pure alcohol showed higher yield compared to alcohol-chloroform solvent. For esterifying 200 mg of wet cells, 2 ml of methanol and 1 ml of ethanol was sufficient to complete in situ transesterification. Under temperatures of 105 °C or higher, 2.5% and 5% concentrations of sulfuric acid was able to successfully convert more than 90% of lipid within 30 min when methanol and ethanol was used as solvents respectively. Also, it was verified that the optimal condition found in small-scale experiments is applicable to larger scale using 2 L scale reactor as well.

Evaluation of various harvesting methods for high-density microalgae, Aurantiochytrium sp. KRS101.

Five technologies, coagulation, electro-flotation (EF), electro-coagulation-flotation (ECF), centrifugation, and membrane filtration, were systematically assessed for their adequacy of harvesting Aurantiochytrium sp. KRS101, a heterotrophic microalgal species that has much higher biomass concentration than photoautotrophic species. Coagulation, EF, and ECF were found to have limited efficiency. Centrifugation was overly powerful to susceptible cells like Aurantiochytrium sp. KRS101, inducing cell rupture and consequently biomass loss of over 13%. Membrane filtration, in particular equipped with an anti-fouling turbulence generator, turned out to be best suited: nearly 100% of harvesting efficiency and low water content in harvested biomass were achieved. With rotation rate increased, high permeate fluxes could be attained even with extremely concentrated biomass: e.g., 219.0 and 135.0 L/m(2)/h at 150.0 and 203.0 g/L, respectively. Dynamic filtration appears to be indeed a suitable means especially to obtain highly concentrated biomass that have no need of dewatering and can be directly processed.

Use of orange peel extract for mixotrophic cultivation of Chlorella vulgaris: increased production of biomass and FAMEs.

Mass cultivation of microalgae is necessary to achieve economically feasible production of microalgal biodiesel, but the high cost of nutrients is a major limitation. In this study, orange peel extract (OPE) was used as an inorganic and organic nutrient source for the cultivation of Chlorella vulgaris OW-01. Chemical composition analysis of the OPE indicated that it contains sufficient nutrients for mixotrophic cultivation of C. vulgaris OW-01. Analysis of biomass and FAME production showed that microalgae grown in OPE medium produced 3.4-times more biomass and 4.5-times more fatty acid methyl esters (FAMEs) than cells cultured in glucose-supplemented BG 11 medium (BG-G). These results suggest that growth of microalgae in an OPE-supplemented medium increases lipid production and that OPE has potential for use in the mass cultivation of microalgae.

Utilization of lipid extracted algal biomass and sugar factory wastewater for algal growth and lipid enhancement of Ettlia sp.

The present study assessed the use of hydrolysate of lipid extracted algal biomass (LEA) combined with the sugar factory wastewater (SFW) as a low cost nutrient and a carbon source, respectively for microalgal cultivation. Microalgal strain Ettlia sp. was both mixotrophically and heterotrophically cultivated using various amounts of hydrolysate and SFW. The culture which was grown in medium containing 50% LEA hydrolysate showed highest growth, achieving 5.26 ± 0.14 gL(-1) after 12 days of cultivation. The addition of SFW increased the lipid productivity substantially from 5.8 to 95.5 mg L(-1)d(-1) when the culture medium was fortified with 20% SFW. Gas chromatography analysis indicated a noticeable increase of 20% in C16 and C18 fraction in FAME distribution under above condition. Therefore, it can be concluded that the combination of LEA hydrolysate and sugar factory waste water can be a powerful growth medium for economical algal cultivation.

Isolation and characterization of thermostable phycocyanin from Galdieria sulphuraria

Phycocyanin is a highly valuable pigmented protein synthesized by several species of cyanobacteria and red alga. In this study we demonstrate the production of thermostable phycocyanin from the unicellular red alga Galdieria sulphuraria. Phycocyanin was extracted by repeated freeze-thaw cycles and purified in a two-step process using ammonium sulfate fractionation, at 25% and 50% concentrations. Purified phycocyanin exhibited maximum absorbance at 620 nm, and the purity ratio (A620/A280) was found to be greater than 4. The recovery efficiency of phycocyanin from the crude extract was above 80%. In total, approximately 19 milligram pure phycocyanin was obtained from 3 g of wet cell mass of Galdieria sp. Subunits α and β of the protein were separated by SDS-PAGE and analyzed by MALDITOF mass spectrometry for identification, which confirmed that the isolated protein is phycocyanin. The molecular weight of α and β subunits of phycocyanin was found to be 17.6 and 18.4 kDa, respectively.

Energy-efficient cultivation of Chlamydomonas reinhardtii for lipid accumulation under flashing illumination conditions

Microalgal cultivation has been limited by the efficiency and costs associated with providing light energy, the most expensive and essential element needed for microalgal growth to a culture, particularly to cultures grown in a photo bioreactor (PBR). This study examined the economic benefits of using flashing illumination conditions in the context of microalgal cultivation. Chlamydomonas reinhardtii was cultivated under various conditions, including various inoculum sizes, light intensities, and durations of the light and dark periods. Our results showed that the highest microalgal growth efficiencies could be obtained using a large inoculum size under high intensity illumination accompanied by a 1:1 ratio of light and dark periods. The duration of the flashing light period was further optimized; permitting light energy savings of 62.5% of the light energy expended under continuous illumination conditions without reducing the biomass or lipid productivity. This study provides a more economical approach to cultivating C. reinhardtii via optimized flashing illumination without sacrificing microalgal growth or lipid content.