Faizal Bux

Bioprospecting for hyper-lipid producing microalgal strains for sustainable biofuel production

Global petroleum reserves are shrinking at a fast pace, increasing the demand for alternate fuels. Microalgae have the ability to grow rapidly, and synthesize and accumulate large amounts (approximately 20-50% of dry weight) of neutral lipid stored in cytosolic lipid bodies. A successful and economically viable algae based biofuel industry mainly depends on the selection of appropriate algal strains. The main focus of bioprospecting for microalgae is to identify unique high lipid producing microalgae from different habitats. Indigenous species of microalgae with high lipid yields are especially valuable in the biofuel industry. Isolation, purification and identification of natural microalgal assemblages using conventional techniques is generally time consuming. However, the recent use of micromanipulation as a rapid isolating tool allows for a higher screening throughput. The appropriate media and growth conditions are also important for successful microalgal proliferation. Environmental parameters recorded at the sampling site are necessary to optimize in vitro growth. Identification of species generally requires a combination of morphological and genetic characterization. The selected microalgal strains are grown in upscale systems such as raceway ponds or photobireactors for biomass and lipid production. This paper reviews the recent methodologies adopted for site selection, sampling, strain selection and identification, optimization of cultural conditions for superior lipid yield for biofuel production. Energy generation routes of microalgal lipids and biomass are discussed in detail.

BODIPY staining, an alternative to the Nile Red fluorescence method for the evaluation of intracellular lipids in microalgae

In order to develop feasible production processes for microalgal biodiesel, the isolation of high neutral lipid producing microalgae is crucial. Since the established Nile Red (NR) method for detection of intracellular lipids has been successful only for some microalgae, a more broadly applicable detection method would be desirable. Therefore, BODIPY 505/515, a lipophilic bright green fluorescent dye was tested for detection of intracellular lipids in Chlorella vulgaris, Dunaliella primolecta and Chaetoceros calcitrans. An optimum concentration of 0.067 μg ml(-1) was determined for lipid staining in the microalgae. Compared to NR, BODIPY 505/515 was more effective in staining microalgae and showed resistance to photobleaching, maintaining its fluorescence longer than 30 min.

PAM fluorometry as a tool to assess microalgal nutrient stress and monitor cellular neutral lipids

This study investigated the use of Pulse Amplitude Modulated (PAM) fluorometry to measure nutrient induced physiological stress and subsequent synthesis of cellular neutral lipids. A freshwater Chlorella sp. was subjected to complete nutrient stress (distilled H2O) and selective nutrient stress in modified BG-11 media (BG-11-N, BG-11-P and BG-11-Fe). Physiological stress was recorded using parameters, rETR, Fv/Fm, Ek, α and NPQ. Induced stress became evident when these parameters were significantly altered, suggesting the onset of neutral lipid synthesis. Complete nutrient stress induced the highest yield of cellular neutral lipids (∼49%) compared to absence of selected nutrients (∼30%). Physiological stress was recorded by a significant decrease in rETR (75%), Fv/Fm (36%), and Ek (60%) and an increase in NPQ (83%). Optimization of neutral lipids occurred by initially maximizing the biomass and subsequently subjecting the harvested biomass to complete nutrient stress.

Effects of parameters affecting biomass yield and thermal behaviour of Chlorella vulgaris.

Conventional fossil fuels are facing a global challenge which lead scientists to explore alternative fuel production from biological sources. The algae-based fuels are gaining rapid attention as it has potential to replace petroleum-based fuels. An indigenous high lipid producing microalgae was isolated from a freshwater pond in the KwaZulu-Natal province of South Africa. The isolate was later identified as Chlorella vulgaris, based on partial 28S large subunit ribosomal RNA gene sequence. The growth kinetics, pyrolytic characteristics and photosynthetic efficiency of Chlorella was evaluated in vitro. The optimized conditions for higher biomass yield of the selected strain were at 4% CO(2), 0.5 g l(-1) NO(3) and 0.04 g l(-1) PO(4), respectively. The pulse amplitude modulation results indicated that C. vulgaris could withstand a light intensity ranging from 150 to 350 μmol photons m(-2)s(-1). Further increase in light intensity resulted in a decline of the electron transport rate. Carbon fixation rate, lipid content and calorific value of C. vulgaris was 6.17 mg l(-1)h(-1), 21% and 17.44 kJ g(-1), respectively. The pyrolitic studies under inert atmosphere at different heating rates of 15, 30, 40 and 50°C min(-1) from ambient temperature to 800°C showed that the overall final weight loss recorded for the four different heating rates was in the range of 78.9-81%. These studies could be useful to appraise the biofuel potential of the isolated C. vulgaris strain, which can later be taken for pilot scale production.

The optimization of biomass and lipid yields of Chlorella sorokiniana when using wastewater supplemented with different nitrogen sources.

The potential of nitrogen sources supplementing domestic wastewater for the cultivation of microalgae was assessed. Urea, potassium nitrate, sodium nitrate and ammonium nitrate were evaluated for their effect on cultivation and lipid production of Chlorella sorokiniana. Urea showed the highest biomass yield of 0.220 g L(-1) and was selected for further experimentation. Urea concentrations (0-10 g L(-1)) were assessed for their effect on growth and microalgal physiology using pulse amplitude modulated fluorometry. A concentration of 1.5 g L(-1) urea produced 0.218 g L(-1) biomass and 61.52% lipid by relative fluorescence. Physiological stress was evident by the decrease in relative Electron Transport Rate from 10.45 to 6.77 and quantum efficiency of photosystem II charge separation from 0.665 to 0.131. Gas chromatography analysis revealed that C16:0, C18:0, C18:1, C18:2 and C18:3 were the major fatty acids produced by C. sorokiniana. Urea proved to be an effective nitrogen supplement for cultivation of C. sorokiniana in wastewater.

Trends in biohydrogen production: major challenges and state-of-the-art developments

Hydrogen has shown enormous potential to be an alternative fuel of the future. Hydrogen production technology has gained much attention in the last few decades due to advantages such as its high conversion efficiency, recyclability and non-polluting nature. Over the last few decades, biological hydrogen production has shown great promise for generating large scale sustainable energy to meet ever increasing global energy demands. Various microorganisms, namely bacteria, cyanobacteria, and algae which are capable of producing hydrogen from water, solar energy, and a variety of organic substrates, are explored and studied in detail. Current biohydrogen production technologies, however, face two major challenges such as low-yield and high production cost. Advances have been made in recent years in biohydrogen research to improve the hydrogen yield through process modifications, physiological manipulations, through metabolic and genetic engineering. Recently, cell immobilization such as microbes trapping with nanoparticles within the bioreactor has shown an increase in hydrogen production. This review critically evaluated various biological hydrogen production technologies, key challenges, and recent advancements in biohydrogen research and development.

Lipid extracted algae as a source for protein and reduced sugar: a step closer to the biorefinery

The objective of this study was to investigate the feasibility of using lipid extracted algae (LEA) as a source for protein and reduced sugar, and the effects of various procedural treatments on their yields. LEA provided comparable yields of protein and reduced sugars to those from total algae. Oven drying provided highest yields of all products followed by freeze drying, while sun drying significantly lowered their yields. Effective cell disruption by microwave and autoclave increased the lipid yields from algae, but resulted in increased loss of other compounds with lipid extracting solvents lowering their yields during sequential extraction. Relatively inefficient cell disruption by ultrasonication and osmotic shock lowered the amount of cell protein lost to the lipid extracting solvents. These results highlight the complexity of concurrent extraction of all value added products from algae, and the need for proper selection of the processes to achieve the objectives of integrated biorefinery.

Application of quantitative RT-PCR to determine the distribution of Microthrix parvicella in full-scale activated sludge treatment systems.

Three wastewater treatment plants in South Africa were investigated to understand the phylogeny and distribution of Microthrix parvicella using real-time polymerase chain reaction (RT-PCR). The phylogenetic analysis of the 16S rRNA of M. parvicella revealed 98% to 100% homology of South African clones to M. parvicella reported in Genbank. The standard curves for RT-PCR showed R2 values greater than 0.99, accurate for quantification. The relative occurrence of M. parvicella 16S rRNA gene copies in the three wastewater treatment plants was in the range 0% to 3.97%. M. parvicella copies increased when the environmental temperature (≤20°C) and food/microorganism (F/M) ratio was low. The M. parvicella 16S rRNA copies could be positively correlated to the sludge volume index at low temperature. At higher temperature, there was a rapid reduction in M. parvicella population irrespective of other favorable factors, indicating the strong influence of temperature on filamentous proliferation. RT-PCR has potential applications in wastewater treatment plants to monitor sudden shift in the microbial population and assessing the plants efficacy.

OVERVIEW OF THE POTENTIAL OF MICROALGAE FOR CO2 SEQUESTRATION

An economic and environmentally friendly approach of overcoming the problem of fossil CO2 emissions would be to reuse it through fixation into biomass. Carbon dioxide (CO2), which is the basis for the formation of complex sugars by green plants and microalgae through photosynthesis, has been shown to significantly increase the growth rates of certain microalgal species. Microalgae possess a greater capacity to fix CO2 compared to C4 plants. Selection of appropriate microalgal strains is based on the CO2 fixation and tolerance capability together with lipid potential, both of which are a function of biomass productivity. Microalgae can be propagated in open raceway ponds or closed photobioreactors. Biological CO2 fixation also depends on the tolerance of selected strains to high temperatures and the amount of CO2 present in flue gas, together with SOx and NOx. Potential uses of microalgal biomass after sequestration could include biodiesel production, fodder for livestock, production of colorants and vitamins. This review summarizes commonly employed microalgal species as well as the physiological pathway involved in the biochemistry of CO2 fixation. It also presents an outlook on microalgal propagation systems for CO2 sequestration as well as a summary on the life cycle analysis of the process.

Improving the feasibility of producing biofuels from microalgae using wastewater

Biofuels have received much attention recently owing to energy consumption and environmental concerns. Despite many of the technologies being technically feasible, the processes are often too costly to be commercially viable. The major stumbling block to full-scale production of algal biofuels is the cost of upstream and downstream processes and environmental impacts such as water footprint and indirect greenhouse gas emissions from chemical nutrient production. The technoeconomics of biofuels production from microalgae is currently unfeasible due to the cost of inputs and productivities achieved. The use of a biorefinery approach sees the production costs reduced greatly due to utilization of waste streams for cultivation and the generation of several potential energy sources and value-added products while offering environmental protection. The use of wastewater as a production media, coupled with CO2 sequestration from flue gas greatly reduces the microalgal cultivation costs. Conversion of residual biomass and by-products, such as glycerol, for fuel production using an integrated approach potentially holds the key to near future commercial implementation of biofuels production.