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Title Page
Table of Contents

1.1       Preamble
1.2       Research Problems
1.3       Research Justification
1.4       Research Aim and Objectives
1.5       Scope of The Study

2.1       Cellulose
2.2       Cellulose Chemistry
2.3       Cellulases
2.4       Cellulose Conversion
2.5       Pre-Treatment Of Lignocellulosic Material
2.5.1    Physical Pre-Treatment
2.5.2    Physicochemical Pre-Treatment
2.5.3    Chemical Pre-Treatment
2.5.4    Biological Pretreatment
2.6       Factors Affecting Enzymatic Hydrolysis Of Cellulose
2.6.1    Enzyme-Related Factors Affecting Hydrolysis
2.6.2    Effects of Substrate Concentration
2.6.3    Effects of Substrate Particle Size
2.6.4    Effects of pH
2.6.5    Effects of Temperature
2.7       Ethanol
2.7.1    Uses of Ethanol
2.7.2    Ethanol as Disinfectants
2.7.3   Ethanol as Domestic Lighting Agent
2.7.4   Ethanol as Transportation Fuel
2.7.5   Other Uses of Ethanol
2.7.6    Chemistry of Bio-Ethanol
2.7.7    Properties of Ethanol
2.7.8    Bio-Ethanol Production Process
2.7.9    Process Description of Simultaneous Saccharification and Fermentation (SSF)
2.7.10 Fermentation
2.7.11  Distillation
2.7.12 Dehydration

3.0 Materials And Method
3.1       List of Materials and Equipment
3.2       Culturing of Aspergillus Niger
3.2.1    Preparation of Culture Medium (Potato Dextrose Agar, PDA)
3.2.2    Pour Plating
3.2.3    Isolation of Organisms
3.2.4    Identification of Fungus
3.2.5    Sub-Culturing
3.3       Sample Collection/Preparation
3.4       Pre-Treatment of Substrate
3.5       Substrate Analysis
3.6       Simultaneous Saccharification And Fermentation (SSF) Of Elephant Grass
3.6.1    Effect of Substrate Concentration
3.6.2    Effect of Substrate Particle Size
3.6.3    Effect of Aspergillus Niger Concentration
3.6.4    Effect of Yeast Concentration
3.6.5    Effect of Temperature
3.6.6    Effect of pH
4.0 Results And Discussion
4.1       Introduction
4.2       Determination of Suitable Pre-Treatment Method
4.3       Isolation of Aspergillus Niger
4.4       Simultaneous Sacharification And Fermentation Of Elephant Grass Stem
4.4.1    Fourier Transform Infrared Analysis (FTIR)
4.4.2    Effect of Pre-Treatment on Ethanol Yield
4.4.3    Effect of Substrate Concentration on Ethanol Yield
4.4.4    Effect of Substrate Concentration on Residual Sugar
4.4.5    Effect of Temperature on Ethanol Yield
4.4.6    Effect of Temperature on Residual Sugar
4.4.7    Effect of Particle Size on Ethanol Yield
4.4.8    Effect of Particle Size on Residual Sugar
4.4.9    Effect of pH on Ethanol Yield
4.4.10  Effect of pH on Residual Sugar
4.4.11  Effect of Cell Loading Effect of Aspergillus Niger Concentration on Ethanol Yield Effect of Yeast Concentration on Ethanol Yield
4.5       Kinetic Parameters

5.0 Conclusions And Recommendations
5.1       Conclusions
5.2       Recommendations


The production of bio-ethanol from Elephant grass (Pennisetun purpureum) stem was carried out using elephant grass stem as a feedstock and a combination of Aspergillus niger at 0.2%(w/v) 0.4%(w/v), 0.6%(w/v), 0.8%(w/v) and 1%(w/v) concentrations and

Saccharomyces cerevisiae (brewer’s yeast) at 0.5% (w/v), 1%(w/v), 1.5%(w/v), 2%(w/v) and 2.5%(w/v) concentrations as cells by simultaneous saccharification and fermentation (SSF). The study determined the most suitable pre-treatment method from the following pretreatment methods; 1M H2SO4, 0.1M H2SO4, 1M NaOH, 0.2M NaOH, Boiling, and 3M NH4OH. IM NaOH pre-treatment gave the highest cellulose and lowest lignin

content. The effects of temperature at 25oC, 30oC, 35oC, 40oC and 45oC; pH values of 3.5, 4.0, 4.5, 5.0, 5.5, 6.0 and 6.5; substrate concentration values of 1%(w/v), 2%(w/v), 3%(w/v), 4%(w/v) and 5%(w/v); particle size range of 53-106µm, 106-150µm, 150-250µm, 250-300µm and 300-425µm; and cell loading combination of Aspergillus niger at 0.2%(w/v) 0.4%(w/v), 0.6%(w/v), 0.8%(w/v), 1%(w/v) concentrations and

Saccharomyces cerevisiae (brewer’s yeast) at 0.5% (w/v), 1%(w/v), 1.5%(w/v), 2%(w/v),

2.5%(w/v) on the hydrolysis and fermentation process were investigated to obtain optimum conditions of fermentation. The optimum conditions of fermentation were obtained at temperature of 350C, pH value of 5.5, substrate concentration of 30g/l, particle size range of 250-300µm and Aspergillus niger to yeast ratio of 0.6/1.5 after 72 hours of fermentation time. Also substrate concentration of 30g/l, gave highest ethanol concentration of 23.4g/l and a yield of 78%. From the research, the kinetic Parameters which are reaction constant k and order of reaction n were evaluated to be 8.172x10-8l/g.s and 2 respectively.



1.1 Preamble

Energy availability, supply and use play a central role in the way societies organize themselves, from individual welfare to social and industrial development. By extension, energy accessibility and cost is a determining factor for the economical, political and social interrelations among nations. Considering energy sources, human society has dramatically increased the use of fossil fuels in the past 50 years in a way that the most successful economies are large consumers of oil. However, geopolitical factors related to security of oil supply, high oil prices and serious environmental concerns, prompted by global warming, the use of petrol for transportation which accounts for one-third of greenhouse gas emissions (Wyman, 1996), have led to a push towards decrease in its consumption. Indeed, the world's strongest economies are deeply committed to the development of technologies aiming at the use of renewable sources of energy. Within this agenda, the substitution of liquid fuel gasoline by renewable ethanol is of foremost importance. Biomass – derived ethanol represents one of the more promising commodities for long term sustainability of transportation fuels (Chum and Overend, 2001).

Brazil has been a front-runner in the use of renewable fuels. The partial substitution of gasoline by ethanol started in 1975. At the time of the first oil crisis, in the 1970s, the country imported 85% of its oil needs, hence the potential for ethanol production from.....

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