Faculty of Applied Sciences
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Item Application of lipid extracted algae in feed and energy production(2019) Ansari, Faiz Ahmad; Bux, Faizal; Gupta, Sanjay Kumar; Guldhe, AbhishekMicroalgae are well considered to be promising feedstocks for biodiesel production. Microalgae can be grown under different types of cultivation conditions and their biomass has tremendous potential to be used as biofuel feedstock and for other applications such as feed, food, cosmetics, pharmaceutical etc. Despite the many benefits and the significant development in the field of microalgal biodiesel production, there are several challenges including high cultivation cost and developing efficient downstream processing methods. The biomass production cost is high, which significantly hinders the use of microalgae as a feedstock. Most of the available literature is focused on upstream, single strain and single product strategy, where mainly algal lipids are used for biofuel production. Hence, for improving the sustainability of the algal biofuel production processes and related process economics, a multiple applications approach using integrated biorefinery and exploiting microalgae for environmental benefits is required. To explore the microalgal biorefinery concept it is vital to understand the various cultivation conditions and applications of biomass in different sectors. There are various strategies, which have potential to make algal biofuel technologies more economically feasible and environmentally sustainable. Use of alternative culture media, improving the biomass production and the efficiency of downstream processing (drying, cell disruption, lipid extraction etc.) algal biofuel technology economical. Utilizing lipid-extracted algae (LEA) for energy and aqua feed application will maximize overall economic return and will leave minimal residues as by-product. The major focus of this thesis was to utilize LEA as substrate for biomethane production and protein source in aquaculture feed. However, effect of preceding steps such as microalgae cultivation, biomass drying and cell disruption on major metabolites extraction was also studied. Microalgae were cultivated in different medium (domestic wastewater and BG11) and their biomass yields and biochemical composition (lipid, protein and carbohydrate) were compared. Different drying and cell disruption techniques were employed for lipid extraction and their effect on lipid, protein and carbohydrate yields were evaluated. The yield of major metabolites on whole cell and LEA were also compared. Suitable solvent systems were selected for optimum lipid extraction from wet and dry biomass with minimal toxic effect on LEA metabolites so that LEA can be further used for biomethane and aquaculture feed production. The choice of microalgae at large scale depends upon the number of factors such as their adaptability to large-scale cultivation, biomass production, major metabolites content, robustness towards the open system cultivation and contamination. In this study, S. obliquus and C. sorokiniana were cultivated in wastewater and BG11 medium at laboratory scale. Both strains are indigenous to KwaZulu-Natal. C. sorokiniana showed lower biomass and major metabolites (lipid, protein and carbohydrate) production at large scale compared to S. obliquus. Considering better adaptability to open cultivation, high biomass and metabolites yields, S. obliquus strain was selected for the LEA application study. Microalgae species, C. sorokiniana and S. obliquus were cultivated on BG11 and using different ratios of raw domestic wastewater and post-chlorinated wastewater as nutrient media. The cultivation of S. obliquus and C. sorokiniana showed biomass yield of 1.2-3.5 and 0.78-1.8 g L-1 in BG11 medium, respectively. While biomass yield observed in wastewater was 0.59-1.59 g L-1 for S. obliquus and 0.67-1.45 g L-1 for C. sorokiniana. The higher biomass yield in BG11 medium attributed to the higher nutrient contents in this medium compared to wastewater. The lipid contents for S. obliquus and C. sorokiniana were 20 and 16.5% dry cell weight (DCW), respectively when grown using BG11 medium. While increases in lipid contents of 26.25 and 29.4% DCW were found for S. obliquus and C. sorokiniana, respectively when cultivated using wastewater. Similarly, carbohydrate contents for S. obliquus and C. sorokiniana were 18 and 17% DCW, respectively for BG11 medium. Increased in carbohydrate contents of 25% for S. obliquus, 28.4% DCW for C. sorokiniana were observed for wastewater. Microalgae tend to accumulate more lipids and/or carbohydrates under nutrient stress condition. The nitrogen and phosphorus contents in wastewater are lower than BG11 medium, which were responsible for stressed condition for microalgae. With limited nutrients in wastewater compared to BG11 medium, growth of microalgae is also lower which resulted in lower protein content. Protein content for S. obliquus and C. sorokiniana in BG11 medium were 37.83-48.8 and 25-35.3% DCW, respectively. The protein contents for S. obliquus and C. sorokiniana in wastewater medium were 16.4-27.29 and 15.8-27.3% DCW, respectively. The biochemical composition depends upon the nutrient composition of the medium and cultivation conditions. The two selected microalgae have shown potential for nutrient removal while cultivated in wastewater. The removal efficiency by S. obliquus was found to be 76.13% for COD, 98.54% for nitrogen and 97.99% for phosphate. Microalgae C. sorokiniana cultivation in wastewater removed 69.38% COD, 86.93% nitrogen and 68.24% phosphates. Increased lipid accumulation in the cells was also recorded in stressed conditions due to low nutrient availability from wastewater. After harvesting of microalgae from culture media, the water content in thick algal slurry (>85% DCW) lowers the products recovery. To overcome this challenge drying and cell disruption are required to enhance the efficiency of lipid extraction. Where drying and cell disruption increase the viability of biomass for lipid extraction process. Three biomass-drying techniques viz. sun, oven and freeze-drying and four-cell disruption techniques viz. microwave, sonication, osmotic shock and autoclave disruption were studied for their effect on recovery of major metabolites from S. obliquus. Microalgae metabolites recovery from whole cell and LEA were analysed and compared. The results showed that after lipid extraction, LEA still contained comparable protein to whole algae biomass however, the carbohydrate concentration was reduced. Oven drying exhibited the highest recovery of all the major metabolites followed by freeze-drying; sun drying however, showed lower yields. Despite lower metabolites recovery sun-drying technique is preferable at large scale due to its easy application and cost-effective nature. The main drawback of sun drying technique is weather dependence and required longer period to dry. The microwave and autoclave microalgal cell disruption improved the lipid yield but loss of other compounds was observed. In osmotic shock treatment, due to poor cell disruption efficiency low lipid were obtained and comparably lower protein loss was noticed during lipid extraction. Lipid extraction is crucial step for microalgae biodiesel production. Solvent-assisted lipid extraction is widely used technique for lipid recovery from dry or wet algae biomass. In a biorefinery approach, it is vital to choose appropriate solvents for the optimum lipid extraction whilst having minimal effect on the remaining metabolites (protein and carbohydrates) in LEA. LEA could be used for energy generation or aquaculture feed applications. Six commonly used organic solvents/ solvent systems were used for lipid extraction from wet and dry biomass. The results showed that the lipid extraction efficiency depends strongly on types of biomass as well as solvent systems selected. Lipid extraction from wet algal biomass could reduce the processing steps and save energy incurred in drying. However, the water present in wet algal slurry acts as a barrier, which results in lower lipid yield compared to the dry biomass. The results revealed that among all six-selected solvents, chloroform: ethanol (1:1 v/v) was most effective if wet biomass used specifically for lipid purpose only. To explore the biorefinery concept, isopropanol/hexane composition is the most suitable solvent system because it is less toxic and resulted in high protein (20.07% DCW) and carbohydrate (22.87%) yields in LEA. For dry algal biomass, chloroform: methanol (2:1 v/v) is an appropriate solvent system if biomass used especially for lipid (19.25%) extraction. If LEA to be used for energy and/or aquaculture feed application, DCM: methanol was found to be a suitable solvent system, which gave 32.79% protein and 26.92% carbohydrate yield. Comparatively hexane has lower lipid recovery but shown higher protein and carbohydrate yield in LEA. Due to less toxic, easy to scale up and inexpensive, hexane is preferable as a solvent for lipid extraction if LEA is to be further utilized at large scale for energy or feed application. Anaerobic digestion (AD) of organic residues is well-researched technology for biomethane production. Whole microalgae and LEA has promising potential for biomethane production. The anaerobic sludge used as inoculum for microalgal biomass digestion. Biomethane production from whole algae and products extracted algae highly depends on sludge to algae biomass ratio for higher methane production. The extraction of metabolites also changes the biochemical composition of residual biomass, which can affect the biomethane production. It is vital to understand the effect of various product-extracted algae and as well as pre-treated algae on the biochemical methane potential. In order to compare biomethane potential, four types of biomass were selected namely sun dried powder algae (SDPA), mild heat-treated algae (MHTA), LEA (using hexane as lipid extracting solvent) and protein-extracted algae (PEA). The average methane (CH4) production rate was ~ 2.5 times higher for protein and lipid extracted algae than for whole algae SDPA and MHTA whilst the cumulative CH4 production was higher for pre-treated algae. Highest cumulative CH4 production (318.7mL CH4 g-1 VS) was found for MHTA followed by SDPA (307.4mL CH4 g-1 VS). The CH4/CO2 ratios of 1.5 and 0.7 were observed for MHTA and LEA, respectively. Outcome of this objective revealed that pre-treatment process disrupts the microalgae cell walls, exposing intracellular material and increasing the surface area. The product-extracted algae changes the elemental composition, which decreases the cumulative gas yield CH4/CO2 ratio. Presence of high nitrogen in the form of protein produces ammonia (NH3) which inhibits the methane production. Therefore, it is imperative to use PEA biomass to improve the methane production yield than the whole cell biomass. Due to escalating price and unstable supply of fish meal (FM), alternative protein sources are used in aqua feed, however these sources do not meet to the requirement. The use of less expensive protein source in aquaculture feed as alternative to FM is required. Microalgae are primary producers in the food chain as well as a natural food for fish. Microalgal biomass is comprised of proteins, lipids, carbohydrates, pigments and many other bioactive compounds. The microalgal proteins have an appropriate balance of all essential amino acids, while lipids are rich in polyunsaturated fatty acids (omega-3 fatty acids, EPA, DHA). Whole algae contain all required ingredients while LEA also contain protein, carbohydrates, vitamins, bioactive compounds even though most of the lipid soluble nutrients have been removed. Thus, microalgae have promising potential to be used in aquaculture feed. Aquaculture production continues to increase globally, to meet the aquaculture feed demand algae supplemented aquaculture feed will play an important role in providing good quality fish. In this study, approximately 200 kg of microalgal biomass was harvested for the feed application. Due to lower toxicity, ease of availability and ease of recovery from mixture, hexane was used as a lipid extracting solvent at pilot scale to generate LEA. The 44 weeks (from juvenile to finisher stage) feeding trials were conducted to evaluate the effect of whole and LEA supplementation of S. obliquus strain on growth performance, disease tolerance, feed utilization, physiological activity, and fillet biochemical composition of Nile tilapia (Oreochromis niloticus). In the first trial, fish were fed with an algae free diet (control) and four experimental diets (2.5, 5, 7.5 and 10 wt%) as protein source of dried S. obliquus. The study showed that microalgae could be used as a protein supplement in the Tilapia feed for enhancement of morphological characteristics and nutritional value. The 7.5% and 10% supplementation of whole algal biomass in tilapia feed showed significant improvement in weight and length of the fish compared to the control. The daily body weight gain was 0.25 g higher in experimental groups than the control. The hepatosomatic index percentage was also higher in fish feed when 7.5% whole algae was used in fish feed as a protein source. The results also showed that 7.5% and 10% have better specific growth rate (1.57 and 1.5%), daily body weight gain (1.1 and 0.86 g), overall body weight gain (427.16 and 331.48 g), protein assimilation (43.96 and 40.46%) higher than the control diet fed fish. The survival rate of fish were 100% at every inclusion level. In second trial (44 weeks), two supplementations (7.5 and 10 wt%) of LEA as protein source were used in Nile tilapia diets. Results showed 7.5% and 10% LEA supplemented feed shown better growth performance than control. The protein content were 42.2%, 41.3% and 36.1% in tilapia fed with 7.5%, 10% LEA and control feed, respectively. The body weight gain, tilapia fed with 7.5% LEA shown 357 g while 10% LEA and control have 331.78 g, and 330.08 gm, respectively. The application of whole and LEA of S. obliquus in tilapia feed, shown appropriate supplementation level for tilapia feed at demonstration scale. This thesis presents advances in knowledge in the field of microalgae biorefinery research for pilot scale operations. This research work has covered various aspects such as effect of drying, cell disruption and lipid extraction on whole and LEA metabolites yield. The extraction of lipid from wet and dry microalgal biomass using various solvent systems provides a new insight for the selection of appropriate solvent systems, which can be used for the large-scale lipid extraction. The study on LEA for biomethane production enhances the understanding about the effects of different pre-treatments and product extractions on biomethane production. The results revealed that the supplementation of whole cell and LEA using S. obliquus for tilapia feed is safe therefore, can be used as an alternative protein source. The findings of this study have both academic and industrial value.Item Production of chitosan and lipids from a newly isolated Mucor circinelloides(2018) Zininga, Johnson Tungamirai; Permaul, Kugen; Singh, SurenFilamentous fungi are well-known sources of a wide variety of industrially-useful biomolecules. This study demonstrates the applicability of a newly isolated oleaginous fungi Mucor circinelloides ZSKP for lipid and chitosan production. Parameters affecting co-production were identified and were statistically optimized, which resulted in a 3–fold improvement in lipid production. The lipid profile showed a high content of unsaturated fatty acids including oleic, linolenic and linoleic acids, while palmitic acid was the major saturated fatty acid (21%). A comparative study to evaluate the efficacy of enzymatic and chemical treatments for biodiesel production from fungal lipids and sunflower oil revealed slightly enhanced production of biodiesel from fungal lipids, using a commercial lipase. The biodiesel synthesized using lipids from M. circinelloides ZSKP satisfied standard specifications and had a higher cetane number (56), lower kinematic viscosity (4.6 mm2/s) and lower acid number (0.03) compared to sunflower oil. Upon optimizing chitosan production and extraction processes the chitosan production was improved 2-fold. The fungal chitosan showed antimicrobial properties and was more effective against Aspergillus niger A chitosan spray was developed which was able to increase the shelf life of fresh fruit produce. These results indicate that Mucor circinelloides ZSKP is a promising candidate for concurrent production of lipids and the versatile bio-polymer chitosan.Item Assessment of microalgal ACCase and rbcl gene expression as a function of nutrient and metal stress(2017) Singh, Poonam; Bux, Faizal; Kumari, Sheena K.; Guldhe, AbhishekMicroalgae are considered to be a potential feedstock for biodiesel production. However, the main concern with regard to the large scale microalgal biodiesel production process is its competence and economic viability. The commercial realization of microalgal biodiesel production requires substantial impetus towards development of efficient strategies to improve lipid yields upstream. Nitrogen (N) and phosphorus (P) stress during cultivation are the widely used lipid accumulation strategies for microalgae. However, these individual nutrient stress strategies are associated with compromised biomass productivity which hampers overall lipid productivity. Lipid enhancement strategies based on light, temperature and CO2 are associated with technological barriers for scale up and incur additional cost. Thus, the main aim of this study was to develop an integrated, easily applicable and scalable lipid enhancement strategy based on nutrients and metals such as N, P, iron (Fe), magnesium (Mg), calcium (Ca) and EDTA stress for selected indigenous microalgal strains. The effect of metal concentrations individually and in combination on microalgal lipids and biomass production is a scarcely exploited area. In this study, a novel approach involving individual as well as combined metals and EDTA stress under N and P limited conditions for lipid enhancement in microalgae was investigated. Microalgal growth physiology, photosynthetic performance, biochemical composition (lipid, carbohydrate and protein) and expression of selected key genes involved in photosynthesis (rbcL) and fatty acid biosynthesis (accD) were studied both under selected individual and combined stress conditions. Out of seven microalgal isolates obtained during the initial isolation and screening process, two strains were selected for lipid enhancement study based on their growth rates, biomass yields, lipid content and lipid productivities. The strains were later identified as Acutodesmus obliquus and Chlorella sorokiniana based on both morphological characteristics and phylogenetical analysis. The selected strains were thereafter subjected to different cultivation conditions involving varying metal, EDTA and nutrient stress conditions. A significant increase in lipid productivity was observed when the concentrations of Fe, Mg and EDTA were increased and Ca was decreased to degree in the N and P stress BG11 medium. For A. obliquus, a highest lipid productivity of 80.23 mgL-1d-1 was achieved with the developed strategy under limited N (750 mg L-1) condition which was 2.18 fold higher than BG11 medium and 1.89 fold higher than N limited condition alone. Similarly, for C. sorokiniana, highest lipid productivity of 77.03 mgL-1d-1 was achieved with the developed strategy under limited N (500 mgL-1) and P (10 mgL-1) which was 2.67 fold higher than BG11 medium and 2.35 fold higher than N and P limited condition alone. For both the microalgal strains, Fe was the most significant trace metal affecting their lipid productivity. These above observations were further confirmed through photosynthetic performance analysis and gene expression studies. At mid log phase, 6.38 and 5.15 fold increases in the expression levels of rbcL gene were observed under combined stress (OCMS+OE) as compared to the control (BG11) condition in A. obliquus and C. sorokiniana respectively. This also resulted in an increased expression level of accD gene involved in lipid biosynthesis to 10.25 fold and 9.79 fold in A. obliquus and C. sorokiniana respectively at late log phase. The results from expression studies of rbcL and accD genes were in compliance with biomass yields, photosynthetic performance, protein yield and lipid productivities for both the strains under different cultivating conditions. The universal applicability of the above strategy was confirmed by applying it to five other microalgae strains isolated in this study which resulted in considerable increase in their overall lipid productivity under optimized conditions. Attempts were made to scale up the lab scale study to open circular pond (3000L) cultivation for A. obliquus. Results showed a 2.08 fold increase in lipid productivity under optimized conditions compared to the control, which emphasizes the scalability of the developed strategy even under uncontrolled conditions. In conclusion, the developed combined metal and EDTA stress strategy not only assisted in alleviating the biomass productivity but also enhanced the lipid accumulation which resulted in overall increased lipid productivity under N and P limited condition. Furthermore, the improved carbohydrate and protein productivities observed with the developed lipid enhancement strategy make it suitable for biorefinery approach with multiple products. An improvement in lipid profile and high biodiesel conversion were also observed with this universally applicable and scalable lipid enhancement strategy confirming their potential applicability during large scale cultivation for biodiesel production.Item The use of PAM fluorometry to assess microalgal physiological stress for the production of biodiesel(2011) White, Sarah Anne; Bux, FaizalUnder environmental stress, most microalgae produce intracellular neutral lipids as a storage mechanism. In the biotechnology industry, these lipids are extracted and converted to microalgal biodiesel; however the extent of the stress is not measured for optimum lipid accumulation. In the series of studies undertaken, Pulse Amplitude Modulated (PAM) fluorometry was used to measure three types of physiological stress on Chlorella sp., a freshwater microalgal species. Biomass and lipid yields were also used as indicators of the induced stress conditions. Firstly, nutrient induced physiological stress and the subsequent synthesis of cellular neutral lipids was investigated.Item Characterization and application of bambara groundnut starch-lipid complexes(2017) Oyeyinka, Samson Adeoye; Amonsou, Eric Oscar; Singh, SurenBambara groundnut (Vigna subterranea) is an indigenous underutilised leguminous crop to Africa. It is a good source of protein and carbohydrate including starch. Bambara groundnut is a traditional crop grown mainly for subsistence in Southern Africa. Bambara groundnut has the advantage of being drought tolerant and can thrive in hot temperatures and poor soil conditions. Therefore, it has great potential as an alternative crop to soya bean and peanuts for cultivation and utilisation. Bambara groundnut starch can potentially be used for various industrial applications. However, native starches are not suitable for most industrial applications, hence the need for modification. Bambara groundnut starch has been previously modified using physical and chemical modification methods. Natural alternatives such as the use of lipids are being sought to modify starches due to the associated risk with chemically modified starch. In this research, Bambara groundnut starch was modified with lipids to improve functional properties, utilisation and application. Specifically, the physicochemical properties of native Bambara groundnut starch obtained from five Bambara groundnut genotypes and three landraces (maroon, brown and cream) were determined. Bambara groundnut starch was modified with lipids (palmitic acid, stearic acid, oleic acid, linoleic acid and lysophosphatidylcholine) and the physicochemical properties of the modified starch were investigated. Further, the influence of high-pressure homogenization on complexation of Bambara groundnut starch with lipids was assessed in comparison with maize and potato starches. Lastly, an application of modified Bambara groundnut starch in biofilm production was also studied. Bambara groundnut landraces generally showed higher amylose contents (approx. 33%) than the genotypes (approx. 28%). Differences were observed in the crystalline patterns of these starches. Bambara groundnut genotypes exhibited the C-type-crystallinity, while the landraces showed the unusual A-type pattern. In terms of functionality, landrace starches showed better swelling than the genotypes. Subsequent studies on modification used maroon Bambara groundnut starch since the amylose content was higher than other landraces and there was a consistent supply of the grains during the period of the study. Generally, Bambara groundnut starch showed higher complexing ability with all the lipids than maize and potato reference samples. These differences in complexing ability among the starches could be due to the variation in amylose contents (Bambara groundnut starch: 31.5%, maize: 22.5% and potato: 24.6%). Fatty acids complexed better with Bambara groundnut starch than lysophosphatidylcholine, which could be due to the structural differences in comparison with the lysophosphatidylcholine molecule. The number of fatty acid in the glycerol backbone and the additional steric hindrance of the polar phosphatidic acid group in the lysophosphatidylcholine may have reduced its complexing ability. Among the fatty acids, palmitic acid complexed better than stearic and the unsaturated fatty acids, possibly due to its short chain length compared to other fatty acids. Bambara groundnut starch showed reduced peak and setback viscosities in the presence of stearic acid, linoleic acid and lysophosphatidylcholine, suggesting the formation of V-amylose complex. Bambara groundnut starch pasted with lipids displayed reduced gelling ability compared to their unmodified counterparts. XRD studies of freeze-dried paste revealed peaks at 2Ѳ = 7.4, 12.9 and 19.9o confirming the formation V-amylose complexes in Bambara groundnut starch. Modification of Bambara groundnut starch with lipids resulted in reduced digestibility. High-pressure homogenization significantly increased the complexing ability of Bambara groundnut starch with lipids. Homogenized Bambara groundnut starch-lipid complexes generally exhibited higher complex index than their unhomogenized counterparts. The higher complexing ability could be attributed to the effect of high-pressure which may have enhanced greater dispersion of lipids in the starch-water system. X-ray diffraction studies also revealed the formation of higher complexes as shown by high intensities at peaks (2Ѳ= 7.4, 12.9 and 19.9o) corresponding to V-amylose complexes. Bambara groundnut starch-lipid complexes displayed significantly higher melting temperatures (95.74-103.82oC) compared to native uncomplexed starch (77.32oC). Homogenized Bambara groundnut starch complexes were non-gelling while the unhomogenized types produced weak gels, with G′ ˃ G″ in the range of 0.1- 10 Hz. Complexation of Bambara groundnut starch with lipids using high-pressure homogenization may be employed in the production of modified starch with non-gelling properties and higher thermal stability suitable for certain industrial application, such as fat replacers in mayonnaise, frozen foods and desserts for a better mouth feel. The physicochemical and mechanical properties of biofilm prepared from Bambara groundnut starch modified with stearic acid at varying concentrations of 0, 2, 4, 6, 7 or 10% were further studied. By SEM, Bambara groundnut starch films containing stearic acid (˃ 2%) showed a progressively rough surface compared to those with 2% stearic acid and the control. The addition of 2% stearic acid to Bambara groundnut starch film reduced water vapour permeability by approximately 17%. However, mechanical properties of starch films were generally negatively affected by stearic acid. Bambara groundnut starch film may be modified with 2% stearic acid for improved water vapour permeability and thermal stability with minimal effect on tensile strength.Item Investigation of combined effect of nitrogen, phosphorus and iron on lipid productivity of microalgae Ankistrodesmus falcatus KJ671624 using response surface methodology(Elsevier, 2015-02-15) Singh, Poonam; Guldhe, Abhishek; Kumari, Sheena K.; Rawat, Ismail; Bux, FaizalEnhancement of lipid accumulation is essential to improve the commercial feasibility of microalgal biodiesel production. An oleaginous microalgal strain, Ankistrodesmus falcatus KJ671624 was evaluated for its potential as a biodiesel feedstock in this study. The collective effect of nutrient (nitrogen, phospho-rous and iron) stresses on the lipid productivity of the selected strain was studied by response surface methodology. The highest lipid content of 59.6% and lipid productivity of 74.07 mg L−1 d−1 was obtained under nutrient stress with nitrogen 750 mg L−1, phosphorus 0 mg L−1 and iron 9 mg L−1. The photosyn-thetic behaviour validates the high lipid productivity under combined nutrient stress condition. Saturated fatty acid composition was increased by 38.49% under selected nutrient stress condition compared to BG11 medium. The enhanced lipid accumulation with suitable lipid profile (C16:0, C18:1, C18:2, C18:3) and biodiesel conversion of 91.54 ± 1.43% achieved in A. falcatus KJ671624 further confirm its potential as a promising feedstock for biodiesel production.Item Biodiesel production from microalgae by enzymatic transesterification(2015) Guldhe, Abhishek; Bux, Faizal; Permaul, Kugen; Mutanda, TauraiMain focus of this study is to investigate the enzymatic-conversion of microalgal lipids to biodiesel. However, preceding steps before conversion such as drying of microalgal biomass and extraction of lipids were also studied. Downstream processing of microalgae has several challenges and there is very little literature available in this area. S. obliquus was grown in the pilot scale open pond cultivation system for biomass production. Different techniques were studied for biomass drying and extraction of lipids from harvested microalgal biomass. Effect of these drying and extraction techniques on lipid yield and quality was assessed. Energy consumption and economic evaluation was also studied. Enzymatic conversion of microalgal lipids by extracellular and whole cell lipase application was investigated. For both applications, free and immobilized lipases from different sources were screened and selected based on biodiesel conversion. Process parameters were optimized using chosen extracellular and whole cell lipases; also step-wise methanol addition was studied to improve the biodiesel conversion. Immobilized lipase was studied for its reuse. Final biodiesel was characterized for its fuel properties and compared with the specifications given by international standards. Enzymatic conversion of microalgal lipids was compared with the conventional homogeneous acid-catalyzed conversion. Enzymatic conversion and chemical conversion were techno-economically investigated based on process cost, energy consumption and processing steps. Freeze drying was the most efficient technique, however at large scale economical sun drying could also be selected as possible drying step. Microwave assisted lipid extraction performed better compared to sonication technique. Immobilized P. fluorescens lipase in extracellular application and A. niger lipase in whole cell application showed superior biodiesel conversion. The extracellular immobilized P. fluorescens lipase showed better biodiesel conversion and yields than the immobilized A. niger whole cell lipase. Both the enzyme catalysts showed lower biodiesel conversion compared to conventional chemical catalyst and higher processing cost. However, techno-economic analysis showed that, the reuse potential of immobilized lipases can significantly improve the economics. Fewer purification steps, less wastewater generation and minimal energy input are the benefits of enzymatic route of biodiesel conversion. Microalgae as a feedstock and lipase as a catalyst for conversion makes overall biodiesel production process environmentally-friendly. Data from this study has academic as well as industrial significance. Conclusions from this study form the basis for greener and sustainable scaling-up of microalgal biodiesel production process.Item An evaluation of the efficacy of using selected solvents for the extraction of lipids from algal biomass by the soxhlet extraction method(Elsevier, 2013-08-14) Ramluckan, Krishan; Bux, Faizal; Moodley, Kandasamy G.The use of solvents for the extraction of lipids from algal biomass has been a method of choice for many years. The soxhlet extraction method was chosen because of its simplicity in operation, relative safety and potential for upscaling to industrial plant level. The source of algal biomass was a raceway pond. Chlorella sp. which is known to produce larger amounts of oil than other indigenous species was used for this investigation. Thirteen solvents spanning a range of polarities and solubilities were selected for this study. Extraction methodology involved the use of single solvents, selected binary solvent mixtures and time-based extractions which were varied from 1 to 5 h. Ultraviolet (UV) spectroscopy was used to determine chlorophyll content of the lipid extracts and gas chromatography was used for the identifica-tion and quantitation of the lipids. Analysis showed that ethanol, chloroform and hexane were generally more efficient in the extraction of lipids than the other solvents studied, producing lipid contents in excess of 10%. The time-based trials indicated that the optimum extraction time was 3 h for the solvents selected. The binary solvent mixture with the greatest extraction efficiency (i.e. >10% lipid extract) was obtained with the 1:1 mixture of chloroform:ethanol. Chlorophyll quantities varied for each solvent extract with chloroform and methanol producing the highest values at >1%. Chromatography was effec-tive in identifying lipids used in the production of biodiesel.