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    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, Faizal
    Enhancement 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.
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    Trends and novel strategies for enhancing lipid accumulation and quality in microalgae
    (Elsevier, 2016) Singh, Poonam; Kumari, Sheena K.; Guldhe, Abhishek; Rawat, Ismail; Misra, Rohit; Bux, Faizal
    In order to realize the potential of microalgal biodiesel there is a need for substantial impetus involving interventions to radically improve lipid yields upstream. Nutrient stress and alteration to cultivation conditions are commonly used lipid enhancement strategies in microalgae. The main bottleneck of applying conventional strategies is their scalability as some of these strategies incur additional cost and energy. Novel lipid enhancement strategies have emerged to research forefront to overcome these challenges. In this review, the latest trends in microalgal lipid enhancement strategies, possible solutions and future directions are critically discussed. Advanced strategies such as combined nutrient and culti-vation condition stress, microalgae–bacteria interactions, use of phytohormones EDTA and chemical additives, improving light conditions using LED, dyes and paints, and gene expression analysis are described. Molecular approaches such as metabolic and genetic engineering are emerging as the potential lipid enhancing strategies. Recent advancements in gene expression studies, genetic and metabolic engineering have shown promising results in enhancing lipid productivity in microalgae; however environmental risk and long term viability are still major challenges.
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    Biodiesel synthesis from microalgae using immobilized Aspergillus niger whole cell lipase biocatalyst
    (Elsevier, 2016) Guldhe, Abhishek; Singh, Poonam; Kumari, Sheena K.; Rawat, Ismail; Permaul, Kugen; Bux, Faizal
    Whole cell lipase catalysis and microalgal feedstocks make overall biodiesel synthesis greener and sustainable. In this study, a novel approach of whole cell lipase-catalyzed conversion of Scenedesmus obliquus lipids was investigated for biodiesel synthesis. Microalgal biodiesel was characterized for its fuel properties. Optimization of process parameters for immobilized Aspergillus niger whole cell lipase-catalyzed biodiesel synthesis was carried out. Highest biodiesel conversion of 53.76% was achieved from S. obliquus lipids at 35 °C, methanol to oil ratio of 5:1 and 2.5% water content based on oil weight with 6 BSPs (Biomass support particles). Step-wise methanol addition was applied to account for methanol tolerance, which improved biodiesel conversion upto 80.97% and gave 90.82 ± 1.43% yield. Immobilized A. niger lipase can be used for 2 batches without significant loss in conversion efficiency. Most of the fuel properties of biodiesel met the specifications set by international standards.
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    Biodiesel production from microalgae by enzymatic transesterification
    (2015) Guldhe, Abhishek; Bux, Faizal; Permaul, Kugen; Mutanda, Taurai
    Main 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.
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    Biocatalytic conversion of lipids from microalgae Scenedesmus obliquus to biodiesel using Pseudomonas fluorescens lipase
    (Elsevier, 2015-01-31) Guldhe, Abhishek; Singh, Bhaskar; Rawat, Ismail; Perumal, Kugen; Bux, Faizal
    Conversion of microalgal lipids using biocatalyst is a novel and greener approach to produce biodiesel. Free and immobilized lipases from Candida sp. and Pseudomonas fluorescens along with free lipases from porcine pancreas and wheat germ were screened for biodiesel conversion of Scenedesmus obliquus lipids. Among selected lipases from various sources immobilized lipase from P. fluorescens showed superior biodiesel conversion. Optimization of reaction parameters viz. lipase amount, temperature, methanol to oil molar ratio and water content was carried out using response surface methodology. Best conversion of 66.55% was achieved at 35 °C, methanol to oil ratio of 3:1 with 10% enzyme amount and 2.5% water content based on oil weight. To tackle methanol tolerance step-wise methanol addition was applied, which improved biodiesel conversion upto 90.81%. Immobilized P. fluorescens lipase can be used for 4 batches without much loss in conversion efficiency (>95%). Biodiesel produced has the cetane number of 51.77, Calorific value of 37.67 MJ kg−1. Most of the fuel properties of biodiesel met the specifications set by ASTM and EN standards.
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    Advances in synthesis of biodiesel via enzyme catalysis : Novel and sustainable approaches
    (Elsevier, 2015-01) Singh, Bhaskar; Mutanda, Taurai; Permaul, Kugen; Bux, Faizal; Guldhe, Abhishek
    Biodiesel, a renewable fuel has a great potential in fulfilling an ever-increasing transport fuel demand. The enzymatic conversion process of feedstock oil to biodiesel is greener when compared to the conventional approach of chemical conversion due to mild reaction conditions and less wastewater generation. Lipases obtained from various microbial sources have been widely applied as catalysts for the conversion of oil to biodiesel. Biodiesel and glycerol obtained by enzymatic conversion have shown a higher purity as compared to that obtained by other conversion techniques. Enzymatic conversion of oil to biodiesel is less energy intensive because of milder reaction conditions and fewer purification steps involved in processing. Lipases, due to their catalytic efficiency and specificity, have emerged as a great tool for converting a wide range of feedstock oils to biodiesel. This manuscript presents an overview of the use of enzymatic conversion for making biodiesel production sustainable and environmentally-friendly. The constraints of enzymatic conversion are the high cost of the enzyme and its inhibition by alcohol and glycerol. The possible solutions to overcome these constraints are discussed. Recent advances to develop an effective process for enzymatic conversion of feedstock oils into biodiesel are critically evaluated. Prospective and challenges in scaling up of this technology are also discussed.