Feasibility of using Chlorella vulgaris for the production of algal lipids, for advancement towards a potential application in the manufacture of commodity chemicals and the treatment of wastewater

Yee Keung Wong, Hong Kong Baptist University

Principal supervisor: Prof. Yung Kin Lam.;Thesis submitted to the Department of Biology.Thesis (Ph.D.)--Hong Kong Baptist University, 2016.


Driven by the increase in industrialization and population, the global demand of energy and material products is steadily growing. Microalgae have come into prominence in the past several decades due to their ability to utilize solar energy to fix atmospheric carbon dioxide, and produce biomass and lipids at productivities much higher than those possible with terrestrial biomass. The main objective of this research is to maximize the biomass and lipid production of Chlorella vulgaris by varying different external conditions so as to achieve the ideal feedstock for the production of commodity chemicals and implement wastewater treatment. The effects of various culture medium compositions on Chlorella vulgaris growth and lipid production were investigated using batch culture. Thirteen culture media: Modified Chu’s No. 10, Bold basal, BG-11, Modified BG-11, N-8, M-8, RM, Modified Spirulina, F-si, Fogg’s Nitrogen free, Fog, F/2, and Johnson medium were compared in terms of optical density, biomass production, specific growth rate and lipid production. Following a 10-day culture in a temperature controlled environment, Bold basal medium was found to have the highest average biomass productivity of 48.056 ± 2.097 mg L -1 day -1 , with overall specific growth rate of (d -1 ): 0.211 ± 0.003 and lipid productivity of 9.295 mg L -1 day -1 among the selected media. This is a basis for the optimization of different cultivating medium to enhance algal lipid production. In order to maximize the quality and quantity of the algal biomass and lipid content in Chlorella vulgaris, different strategies were used using different ratios of nitrogen and phosphorus source in the modified Bold basal medium (BBM). In the 12-day batch culture period, the highest biomass productivity obtained was 72.083 mg L -1 day -1 under Bold basal medium with Nitrogem control Phosphorus limited conditions. The highest lipid content, lipid concentration and lipid productivity obtained were 53.202%, 287.291 mg/L and 23.449 mg L -1 day -1 respectively, under Bold basal medium with Nitrogen Control Phosphorus Deprivation conditions. Nitrogen starvation was found to be the critical factor affecting the biomass production and lipid accumulation while the starvation of phosphorus induced a higher total lipid content and affected the lipid composition of Chlorella vulgaris cultures. Recently, as the demand for pure microalgae strains for the production of algal lipid as a feedstock of renewable energy has been increasing, the designation of an effective photobioreactor (PBR) for mass cultivation is essential to assure stability in the amount of feedstock. Various PBRs design such as bubbling, air-lift, porous air-lift was compared. In general, the bubbling design is a better PBR designs than the others, having the highest biomass concentration of 0.78 g/L during the culture time. Besides, it was observed that the 35 cm draft tube of the porous air-lift PBR had shorter mixing time (24.5 seconds) and higher biomass concentration (0.518 g/L) than the 50 cm air- lift design. The bubbling PBR with the highest gas flowrate of 2.7 L/min produced the highest biomass production of 0.74 g/L within the cultivation time. The information is shown to be a useful guide for determining the optimal condition of the PBRs. Light wavelengths and intensities were determinant factors in affecting the growth and lipid content of autotrophic organisms such as C. vulgaris. The experiment investigated the effect of algal lipid production by using LEDs (Light Emitting Diodes) with different wavelengths. C. vulgaris was grown in the effluent for 10 days under the photoperiod of 18:6 h Light/Dark cycles with different visible light sources (cool white, blue and red) and intensities (50 μmol m -2 s -1 ) at 25°C. The overall maximum dry biomass of 1353.33 mg/L was observed at 50 μmol m -2 s -1 cool white light during 10th culture day, with the highest overall productivity of dry biomass production (117.23 mg/L d -1 ) within cultivation time. The highest lipid content (34.06 %) was obtained with the blue color due to light efficiency and deep penetration to the photosynthetic pigments (chlorophyll) in C. vulgaris. However, the highest lipid productivity was observed in cool white light of 318.63 mg/L during the 10th culture day. The effect of light intensity toward the lipid productivity was further investigated by increasing the light intensity of cool white light. The highest lipid productivity was observed at 110 μmol m -2 s -1 in a light intensity of 658.99 mg/L during the 10th culture day. In high irradiance (110 μmol m -2 s -1 ), the proportion of poly unsaturated fatty acid (C18:1 and C18:2) contributed most of the fatty acid methyl ester (FAME) content in the collected sample, irrespective of all treatments. The next study optimized the harvesting rate of algae by using an electro- coagulation-flotation (ECF) harvester, which combines the electrochemical reaction in the electrodes and the dispersion of hydrogen gas to allow floatation of microalgae cell for surface harvesting. The response surface methodology model (RSM) was employed to optimize different ECF parameters: electrode plate material, electrode plate number, charge of electrode, electrolyte concentration and pH of the solution. The model revealed that aluminum was the best electrode material for the ECF process. It was also found that a three electrode plates setup with one anode and two cathodes had the best performance for harvesting. Additionally, sodium chloride (NaCl) at 8 g/L in harvesting medium could increase the flocculant concentration and reduce electric power consumption. Also, having the culture medium at pH 4 also had a significant effect on improving the flocculant production. Combining these optimal conditions, the highest flocculant concentration reached 2966 mg/L in 60 mins; a 79.8% increase in flocculant concentration, based on the tested conditions. The results of this study show the significance of different parameters affecting the coagulation and flocculation of C. vulgaris and provide a reference for the design of a large-scaled harvester for microalgae harvesting in the further study. To conclude, this research comprises a study on the use of indigenous algae for the production of algal lipid, which is used to produce commodity chemicals. Details on the use of nutrient sources, the techniques of cultivation and the optimization of cell harvesting were included so as to remove nutrients from effluents to minimize the occurrence of eutrophication in harbor, thereby providing economic advantages. Thus, the optimization of these processes is very adequate and offers significant advantages for the wastewater treatment. The developing of algal cell biotechnology is necessary to further enhance algal lipid production in an attempt to apply it commercially.