Min S. Park

Advances in direct transesterification of algal oils from wet biomass

An interest in biodiesel as an alternative fuel for diesel engines has been increasing because of the issue of petroleum depletion and environmental concerns related to massive carbon dioxide emissions. Researchers are strongly driven to pursue the next generation of vegetable oil-based biodiesel. Oleaginous microalgae are considered to be a promising alternative oil source. To commercialize microalgal biodiesel, cost reductions in oil extraction and downstream biodiesel conversion are stressed. Herein, starting from an investigation of oil extraction from wet microalgae, a review is conducted of transesterification using enzymes, homogeneous and heterogeneous catalysts, and yield enhancement by ultrasound, microwave, and supercritical process. In particular, there is a focus on direct transesterification as a simple and energy efficient process that omits a separate oil extraction step and utilizes wet microalgal biomass; however, it is still necessary to consider issues such as the purification of microalgal oils and upgrading of biodiesel properties.

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).

Concurrent extraction and reaction for the production of biodiesel from wet microalgae.

This work addresses a reliable in situ transesterification process which integrates lipid extraction from wet microalgae, and its conversion to biodiesel, with a yield higher than 90 wt.%. This process enables single-step production of biodiesel from microalgae by mixing wet microalgal cells with solvent, methanol, and acid catalyst; and then heating them in one pot. The effects of reaction parameters such as reaction temperature, wet cell weight, reaction time, and catalyst volume on the conversion yield are investigated. This simultaneous extraction and transesterification of wet microalgae may enable a significant reduction in energy consumption by eliminating the drying process of algal cells and realize the economic production of biodiesel using wet microalgae.

An effective, cost-efficient extraction method of biomass from wet microalgae with a functional polymeric membrane

For energy-efficient extraction of biomass from microalgae, it is essential to extract the intracellular lipid directly from wet microalgae without drying the microalgal biomass. In this work, a novel, highly efficient cell disruption process was devised using a functional membrane coated with a cationic polymer. The proposed mechanism of cell disruption involves the perturbation of the local electrostatic equilibrium of the amphiphilic microalgal cell membrane caused by the direct contact with the tertiary-amine cations on the surface of the membrane. A tert-amine-containing polymer, poly-dimethylaminomethylstyrene (pDMAMS) film was conformally deposited on a nylon membrane by a vapor-phase polymerization process, termed as initiated chemical vapor deposition (iCVD). For the wet extraction with this membrane, the pDMAMS-coated membrane was immersed in a microalgal culture of Aurantiochytrium sp. KRS101. The microalgal culture was simply shaken together with the membrane to prompt the contact with the pDMAMS-coated membrane. With this ultimately simple procedure, the bursting of cells was clearly observed. Surprisingly, by this simple, energy-efficient process, a significantly high disruption yield of 25.6 ± 2.18% was achieved. The membrane-based extraction process is highly desirable in that (1) the process does not require an energy-consuming drying procedure, and (2) the proposed cell disruption method with a functional membrane is extremely simple and highly efficient.

Water use and its recycling in microalgae cultivation for biofuel application.

Microalgal biofuels are not yet economically viable due to high material and energy costs associated with production process. Microalgae cultivation is a water-intensive process compared to other downstream processes for biodiesel production. Various studies found that the production of 1 L of microalgal biodiesel requires approximately 3000 L of water. Water recycling in microalgae cultivation is desirable not only to reduce the water demand, but it also improves the economic feasibility of algal biofuels as due to nutrients and energy savings. This review highlights recently published studies on microalgae water demand and water recycling in microalgae cultivation. Strategies to reduce water footprint for microalgal cultivation, advantages and disadvantages of water recycling, and approaches to mitigate the negative effects of water reuse within the context of water and energy saving are also discussed.

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.

Enhancement of lipid productivity by ethyl methane sulfonate-mediated random mutagenesis and proteomic analysis in Chlamydomonas reinhardtii

Microalgae-derived biomass has been considered as the most promising candidate for next generation biofuel due to its sustainability and biodegradability. In this study, microalgal strain Chlamydmonas reinhardtii was randomly mutagenized by using a chemical mutagen, ethyl methane sulfonate (EMS) to create mutants showing enhanced lipid production. We identified three random mutants that displayed high lipid production in the screening using Nile red staining. Among those, mutant #128 was selected as candidate for further studies. Our flow cytometry and confocal microscopy analysis revealed that mutant #128 contains larger and more abundant lipid bodies than that of wild-type. Moreover, mutant #128 showed 1.4-fold increased fatty acid methyl ester (FAME) content compared to wild-type under nitrogen depleted condition. In addition, mutant #128 grew faster and accumulated more biomass, resulting in high lipid production. 2D gel electrophoresis and MALDI-TOF analysis used for gene targeting revealed that β-subunit of mitochondrial ATP Synthase and two-component response regulator PilR may be involved in enhanced characteristics of mutant #128. These results show the possibilities of EMS mediated random mutagenesis in generation of mutants to produce high amount of lipid as well as further study for molecular mechanism of mutants.

A comprehensive study on algal–bacterial communities shift during thiocyanate degradation in a microalga-mediated process

Changes in algal and bacterial communities during thiocyanate (SCN(-)) decomposition in a microalga-mediated process were studied. Pyrosequencing indicated that Thiobacillus bacteria and Micractinium algae predominated during SCN(-) hydrolysis, even after its complete degradation. Principal components analysis and evenness profiles (based on the Pareto-Lorenz curve) suggested that the changes in the bacterial communities were driven by nitrogen and sulfur oxidation, pH changes, and photoautotrophic conditions. The populations of predominant microalgae remained relatively stable during SCN(-) hydrolysis, but the proportion of bacteria - especially nitrifying bacteria - fluctuated. Thus, the initial microalgal population may be crucial in determining which microorganisms dominate when the preferred nitrogen source becomes limited. The results also demonstrated that microalgae and SCN(-)-hydrolyzing bacteria can coexist, that microalgae can be effectively used with these bacteria to completely treat SCN(-), and that the structure of the algal-bacterial community is more stable than the community of nitrifying bacteria alone during SCN(-) degradation.

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.