HYDROLYSIS OF SEAWEED FOR BIOETHANOL PRODUCTION
By
CLAUDIA GITA SUTARDHIO
A Bachelor’s Thesis Submitted to the Faculty of
LIFE SCIENCES
in partial fulfillment of the requirements for the Degree of
BACHELOR OF SCIENCE
WITH A MAJOR IN FOOD TECHNOLOGY
SWISS GERMAN UNIVERSITY Campus German Centre Bumi Serpong Damai – 15321
Island of Java, Indonesia www.sgu.ac.id
July 2009
STATEMENT BY THE AUTHOR
I hereby declare that this submission is my own work and to the best of my knowledge, it contains no material previously published or written by another person, nor material which to a substantial extent has been accepted for the award of any other degree or diploma at any educational institution, except where due acknowledgement is made in the thesis.
_______________________________________ ________________
Claudia Gita Sutardhio Date
Approved by:
________________________________________ __________________
Dr. Ir. Bagus S.B. Utomo, MappSc, Advisor Date
______________________________________ _________________
Dr. rer-nat Filiana Santoso, Co-Advisor Date
______________________________________ _________________
Chairman of the Examination Steering Committee Date
ABSTRACT
HYDROLYSIS OF SEAWEED FOR BIOETHANOL PRODUCTION
By
Claudia Gita Sutardhio
SWISS GERMAN UNIVERSITY Bumi Serpong Damai
Dr. Ir. Bagus S.B. Utomo, MAppSc, Advisor Dr. rer-nat Filiana Santoso, Co-Advisor
This research was divided into two sections. In the preliminary research three cultivation mediums (Nutrient Broth, Specific Cellulose Broth, and Sargassum Broth) were studied to verify the optimum condition for carboxymethylcellulase (CMC-ase) production by Pseudomonas fluorescens. It was found that the highest CMC-ase activity of P. fluorescesns was obtained from non-centrifuged Sargassum Broth at day 8. The objective of the primary research was to determine the best reaction conditions for enzymatic hydrolysis of seaweed with the aim to obtain glucose that will be further converted to bioethanol. The effect of various pH, temperature, and reaction time in the hydrolysis process was studied. The experiment data showed that the highest yield of glucose was achieved at pH 6, 35oC, and 40 minutes reaction time. The hydrolysis under optimum condition produced 1.385 ± 0.064 mg/ml glucose or 4.29% of seaweed by weight. Provided that the conversion of glucose to bioethanol is 90%, the theoretical bioethanol production would be 1.93%
of the seaweed by weight, or in other words, one ton of seaweed would produce 25 L bioethanol. From this research, it is concluded that seaweed could be used to produce bioethanol as renewable energy resource.
DEDICATION
For the world’s greatest parents, Hendra and Nadia, And my beloved sister,
Cindy
who always support, help and motivate me at any time and in any situation
First of all, I want to thank to God for His guidance during my thesis work and His blessing that I can complete this thesis on time.
It is an obligation for all eighth semester students of Swiss German University (SGU) to write a thesis as partial fulfillment of the requirements for the Bachelor Degree.
I realize that to finish this thesis, I get guidance and assistance from others. In this opportunity, I would like to extend my gratitude and appreciation to:
1. Dr. Ir. Bagus S.B. Utomo, MAppSc, as Thesis Advisor, for his great help, assistance, guidance and advice in completing this thesis.
2. Dr. rer-nat Filiana Santoso, as Thesis Co-Advisor, for her great help, assistance, guidance and advice in completing this thesis.
3. Prof. (ris.) Hari Eko Irianto, Ir. PhD., who allows me to do my thesis work at Research Center for Marine and Fisheries Product Processing and Biotechnology (Balai Besar Riset Pengolahan Produk dan Bioteknologi Kelautan dan Perikanan), Jakarta.
4. Balai Besar Riset Pengolahan Produk dan Bioteknologi Kelautan dan Perikanan, which gives me facilities and financial support during completing the thesis.
5. Dr. Maruli Pandjaitan, as the Dean of Life Sciences, Swiss German University (SGU), Serpong, Indonesia, who supports me during completing this thesis.
6. Beloved Parents Hendra and Nadia, and beloved Sister, Cindy, who support me, both in material and non material, during my thesis work.
7. Dr. Singgih Wibowo and Mr. Haryanto, who helps me to do my thesis work in Balai Besar Riset Pengolahan Produk dan Bioteknologi Kelautan dan Perikanan, Jakarta.
8. Ms. Ifah Munifah, Mr. Sugiyono Ronosentono, Ms. Nuri Fawzya, Mr. Tommy, Ms. Novi, and all laboratories personals who give a lot of information and help me in to completing this thesis.
9. Mr. Tabligh Permana, who gives me advices and a lot of information during my thesis work.
10. Mr. Dede K. Saputra, Mr. Irvan Kartawiria, Mr. Tutun Nugraha, Mr. Hery Sutanto, Ms. Mina Arsita, and Mr. Agung Margiyanto who support me during completing this thesis.
11. Rivanny Rinaldy, Selvienna, Yessika Anggraika, Vicky Lysandra, Fitria Cahyadi, Herando Susanto, Lingsia, as Author’s friend, for their great help and support.
12. All Life Sciences Students Swiss German University, especially those who gives me motivation and the biggest support.
13. All E.Nopi Puri Kencana Instructors, especially those who gives me motivation and the biggest support.
14. Some other people that I can not mention one by one who have made some contribution in completing this thesis.
I realize that this thesis is still far from perfect. Therefore, any comments or critics will be welcomed in order to improve this internship report.
I hope that this thesis can give benefits and contributions for any academic purposes, especially Food Technology Faculty, readers and general society.
Jakarta, July 2009
Claudia Gita Sutardhio
TABLE OF CONTENTS
Page
STATEMENT BY THE AUTHOR……….……..2
ABSTRACT………...3
DEDICATION………...4
ACKNOWLEDGEMENTS………...5
CHAPTER 1 INTRODUCTION……….15
1.1 Background………... 15
1.2 Research Problem……….16
1.3 Research Objectives………. ………...16
1.4 Hypothesis………....17
1.5 Research Scope……… 17
CHAPTER 2 LITERATURE REVIEW……….. 18
2.1 Seaweed………...20
2.1.1 Brown Seaweed………21
2.1.2 Sargassum sp..……… 25
2.1.3 Polysaccharide Component in Seaweed.……… 25
2.2 Pseudomonas fluorescence……….. 28
2.2.1 General Characteristic of P. fluorescens……….. 29
2.2.2 Useful function of P. fluorescens……….30
2.2.3 Growth of P. fluorescens………. 31
2.2.4 P. fluorescens as Cellulase Producing Bacteria……….. 32
2.3 Hydrolysis………... 38
2.3.1 Enzymatic Hydrolysis vs Acid Hydrolysis………. 38
2.3.2 Cellulase in Enzymatic Hydrolysis of Cellulose………. 38
2.3.3 Hydrolysis of Biomass for Bioethanol Production………...42
2.4 Bioethanol……….44
2.4.1 Benefits of Bioethanol Utilization………44
2.4.2 Application of Bioethanol in Replacing Fossil Fuels………….. 45
2.4.3. Bioethanol vs Biodiesel………46
2.4.4. Bioethanol Development in the World……… 47
2.4.5. Problems in First Generation of Bioethanol……… 49
2.4.6. Second Generation Bioethanol from Cellulosic Material ………50
CHAPTER 3 RESEARCH METHODOLOGY……….. 52
3.1 Time and Venue……….. 52
3.2 Materials………... 52
3.2.1. Raw materials……….. 52
3.2.2. Chemicals……….53
3.3 Equipments………... 54
3.4 Procedures……… 55
3.5 Experimental Design……… 60
CHAPTER 4 RESULTS AND DISCUSSION……… 62
4.1 Preliminary Studies……….. ………62
4.1.1. Characterization of Sargassum sp………….62
4.1.2. Characterization of Pseudomonas fluorescens……….63
4.1.2.1. General Characteristic of P. fluorescens………. ………63
4.1.2.2. Qualitative Determination of CMC-ase Activity……… 64
4.1.2.3. Quantitative Determination of CMC-ase Activity…….. 65
4.1.3. Influence of Centrifugation Process on CMC-ase Activity of P. fluorescens………71
4.2. Hydrolysis of Sargassum sp. using Cellulase of P. fluorescens……… 73
4.3. Prediction of Bioethanol Production……….. 77
CHAPTER 5 CONCLUSIONS AND RECOMMENDATIONS………... 78
5.1 Conclusions………... 78
5.2 Recommendations………... 78
GLOSSARY………. 79
LIST OF REFERENCES………. 81
APPENDICES………... 90
CURRICULUM VITAE……… 126
