FLOW CULTURE OF SCENEDESMUS DIMORPHUS IN FILTERED SAGO EFFLUENT Nur Jannah Monib This project is submitted in partial fulfillment of the requirements for the degree of Bachelor of Science with Honours (Resource Biotechnology) Faculty of Resource Science and Technology (UNIVERSITI MALAYSIA SARAWAK) 2010
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FLOW CULTURE OF SCENEDESMUS DIMORPHUS IN FILTERED SAGO
EFFLUENT
Nur Jannah Monib
This project is submitted in partial fulfillment of the requirements for the degree of Bachelor of
Science with Honours
(Resource Biotechnology)
Faculty of Resource Science and Technology
(UNIVERSITI MALAYSIA SARAWAK)
2010
i
ACKNOWLEDGEMENTS
Firstly, I would like to express my utmost gratitude to Al-Mighty Allah S.W.T for giving me the
opportunity, strength, and guidance to complete this study. I would also like to express my
sincere gratitude to many people in the course of undertaking this project.
Secondly, I would like to express my sincere thank to my supervisor, Prof. Dr. Kopli Bujang for
his support, guidance and inspiration. I also would like to thank Dr. Cirilo Nolasco Hipolito for
his constant encouragement, guide and assistance, to Miss Rubena Kamal, Mr. Ugam ak Janggu
and M.Sc. students of the Resource Biotechnology Program, UNIMAS for their inspiration and
occasional help and also to our lab assistants who helped to accomplish this project. Last but not
least, I would like to thank my parent and my friends for their support and encouragement.
ii
TABLE OF CONTENTS
Acknowledgements i
Tables of Contents ii
List of Abbreviations v
List of Tables vi
List of Figures vii
Abstract viii
CHAPTER 1: INTRODUCTION
1.1 General Introduction 1
1.2 Objectives 3
CHAPTER 2: LITERATURE REVIEW
2.1 Microalgae 4
2.1.1 Scenedesmus dimorphus 4
2.1.1.1Morphology and characteristic 4
2.1.1.2 Cultivation of Scenedesmus dimorphus 5
2.1.1.2.1 Influence of temperature 6
2.1.1.2.2 Effect of light 6
2.1.1.2.3 pH of medium 6
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2.1.1.2.4 Mixing or agitation of culture 7
2.2 Sago effluent 7
2.2.1. Sago Palm 7
2.2.2 Sago effluent 8
CHAPTER 3: MATERIALS AND METHODS
3.1 Microorganism 9
3.2 Inoculum 9
3.3 Sago effluent 9
3.4 Characterization of sago effluent 10
3.4.1 Total Suspended Solid (TSS) 10
3.4.2 Analysis of Reducing Sugar 11
3.4.3 Starch Analysis 11
3.5 Chu medium 12
3.6 Lab-scale culture 14
3.7 Sampling 15
3.8 Lipid Extraction 16
3.8.1 Harvesting of cells 16
iv
3.9.2 Drying and grinding of cells 16
3.9.3 Lipid Extraction using Soxhlet 16
3.9.4 Evaporation in Rotary evaporator 17
CHAPTER 4: RESULTS AND DISCUSSIONS
4.1 Characterization of sago effluent before filter 19
4.2 Cultivation of Scenedesmus dimorphus 21
4.3 Cultivation of Scenedesmus dimorphus in Filtered Sago Effluent (FSE) 23
4.2.1 Effects of Sodium Bicarbonate (NaHCO3) on the cell growth 23
4.2.2 Glucose contents in the cultivation 26
4.2.3 Starch contents in the cultivation 28
4.3 Lipid Extraction 29
CHAPTER 5: CONCLUSION AND RECOMMENDATIONS 30
References 31
Appendices
v
LIST OF ABBREVIATIONS
DCW dry cell weight
g gram
g/L gram per liter
L liter
lux luxury
mg miligram
ml milliliter
FSE Filtered sago effluent
OD optical density
rpm rotation per minute
% percent
°C degree Celsius
vi
LIST OF TABLES
Table 3.1: Modified Bristol’s Medium
Table 3.2: Chu stock solution
Table 3.3: Trace mineral solution (x)
Table 4.1: Characteristics of sago effluent
Table 4.2: Lipid contains in Scenedesmus dimorphus cultured in FSE
TABLE 1: Modified Bristol’s Medium
TABLE 2: Bristol Medium
vii
LIST OF FIGURES
Figure 2.1: (a) Microscopic picture of Scenedesmus dimorphus at 1000x
Figure 2.1: (b) Microscopic picture of Scenedesmus dimorphus under NIES-93
Figure 3.1: Scenedesmus dimorphus in Proteose medium
Figure 3.2: Cultivation in flow culture using clear Perspex boxes
Figure 3.3: Lipid Extraction
Figure 3.4: Flow chart of obtaining lipid from Scenedesmus dimorphus cultured using filtered
sago effluent (FSE)
Figure 4.1: Scenedesmus dimorphus growth in FSE and Chu medium
Figure 4.2: Glucose contents in the cultivation
Figure 4.3: Starch contents in the cultivations
FIGURE 1: Flow diagram of filtered sago effluent
FIGURE 2: Flow diagram of DNS method (Miller, 1959)
FIGURE 3: Flow diagram of analysis of starch
FIGURE 4: Glucose standard curve detected at 575 nm
FIGURE 5: Starch standard curve detected at 590 nm
viii
Flow Culture of Scenedesmus Dimorphus in Filtered Sago Effluent
Nur Jannah Monib
Resource Biotechnology Program
Faculty of Resource Science and Technology
Universiti Malaysia Sarawak
ABSTRACT
In this study, Scenedesmus dimorphus, photosynthetic green algae was cultivated in filtered sago effluent that added
with NaHCO3 and compared with the filtered sago effluent without NaHCO3 to investigate the effect of the
NaHCO3 to the cultivation of Scenedesmus dimorphus. The cultivation was done for 20 days in the flow culture
with the speed flow 12.3 ml/sec under natural light. After the cultivation, the lipid extraction was done to determine
the lipid content of Scenedesmus dimorphus. The result showed that the cultivation in FSE without NaHCO3
produced the highest dry cell weight which is 395 mg/L on day 10 of cultivation. The cultivation in FSE contain
4g/L produced 315 mg/L. There are another factor that can affect the cultivation of Scenedsmus dimorphus in FSE
such as pH of medium, light and agitation. In addition, the highest lipid content of Scenedesmus dimorphus that was
obtained in this study is 18.39%. Cultivation of Scenedsmus dimorphus in filtered sago effluent can be improved by
providing optimum conditions such as nutrients, light, agitation and environment condition for the cultivation of this