THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY SRIWIJAYA UNIVERSITY SANGVONE SOULIYA BACHELOR THESIS Study Mode : Full-time Major : Environmental Science and Management Faculty : International Programs Office Batch : K45 AEP (2013-2017) Thai Nguyen,22/10/ 2017 Thai Nguyen University of Agriculture and Forestry Degree Program Bachelor of Environmental Science and Management Student Name Sangvone Souliya Student ID DTN1454290077 Thesis Title ―Bioethanol production from Sugarcane Bagasse by Separate Hydrolysis and Fermentation using YEAST ISOLATED FROM DURIAN ((Durio zhibetinus) FRUIT‖ Supervisor (s) 1. Nguyen Huu Tho,Ph. Supervisor‘s signature (s) Abstract: Sugarcane bagasse is the most abundant agriculture waste in the world. It is an attractive feedstock for large-scale biological production of bioethanol.
Therefore, production of ethanol (bioethanol) from lignocellulosic biomass is one way to reduce environment pollution and crude oil from fossil. Bioethanol can produce biofuel by fuel in gasoline engine mixed with ethanol. Bioethanol production from sugarcane bagasse can produce with three main steps by pretreatment, hydrolysis and fermentation. In this study Pretreatment or delignification conducted physical method using steam autoclave for 1 hour to remove lignin.
Followed by hydrolysis in addition of 5% in each samples to dilute or concentration acid hydrolysis. Fermentation used Na-OH to measure ethanol concentration was obtained pH 4. The results showed that the highest of ethanol production was 0.041ml during 30 minutes hydrolysis timed and fermentation process by shaking incubator was 8 days. Keywords: Bioethanol production; Biofuel; Feedstock; Lignocellulosic biomass; Pretreatment; Hydrolysis; Fermentation ii Number of 44 pages Date of October, 2017 Submission iii ACKNOWLEDGMENT I gratefully acknowledge the support and guidance of faculty members of Chemistry Department of Mathematic and Natural Science Faculty in Sriwijaya University, Indonesia.
Most particularly I would like to express my deep thanks to my supervisors Hermansyah,Ph. of Chemistry Department of Mathematic and Natural Science Faculty in Sriwijaya University, Indonesia and Nguyen Huu Tho,Ph. of Department of Science Management and International Relation, Thai Nguyen University of Agriculture and Forestry (TUAF), Vietnam who so kindly participated in this research by giving their generously of their time. Without their thoughtful encouragement and careful supervision.
This thesis would never have taken shape. I am also deeply thankful to Mr. Deddy and Prof Aldes from research laboratory and all of my friends from Indonesia for giving me very kindly and helpful during doing the experiment. Finally, I would like to express my deepest thanks from my heart to my family for their very supportive in every way and all good friends that beside me for along.
iv TABLE OF CONTENT List of Figure………………………………………………………………….vii List of Tables……………………………………………………………….…viii List of Abbreviations………………………………………………….3 Research questions and hypotheses .2 Renewable energy or alternative energy .5 Processing of lignocellulosic to ethanol .4 Separate hydrolysis and fermentation.2 List of apparatus.3 List of reagents.1 Processing of lignocellulosics to bioethanol.1 Preparation of fermentation medium.2 Preparation of YPD agar medium .3 Preparation of YPD broth. RESULT AND DISCUSSION .3 Ethanol analysis curve. 44 vi LIST OF FIGURES Figure 1 Classification of biofuels. 9 Figure 2 Framework of Ethanol Fermentation Process…………………….17 Figure 3 Preparation of sample.
21 Figure 4 A laboratory being used for the fermentation culture medium…. 23 Figure 5 Preparation of inoculum growth cell.24 Figure 6 Diagram of ethanol standard curve. 27 Figure 7 Comparison ethanol analysis with different hydrolysis time. 30 Figure 8 Standard curve (reducing glucose).
31 Figure 9 The curve of reducing sugar in three different hydrolysis time….32 vii LIST OF TABLES Table 1 The difference hydrolysis time. 22 Table 2 Absorbance of glucose analysis by DNS. 25 Table 3 The correlation between the concentration of glucose at various levels and the value Absorbance derived from glucose analysis by DNS. 33 Table 4 Analysis of ethanol on different incubation time.
32 viii LIST OF ABBREVIATIONS C - carbon - carbon dioxide DNS - Di-Nitro Salicylic Acid g - gram g/L - gram per liter - methane H – hydrogen O - Oxygen Mg- Magnesium min - minutes mL - milliliter mg - milligram Na – sodium OH - Hydroxide % - Percentage ix °C - degree celsius μL – microliter n – a letter that represents an unknown number - water EtOH – ethanol – Sulfate radical – ammonium GHG- Greenhouse gases SHF – Separate Hydrolysis and Fermentation SSF- Simultaneous Saccharification and Fermentation YPD or YEPD- Yeast extract peptone dextrose x PART I. Research rationale Lao PDR is an agricultural based country and it is the most important economic sector of Lao PDR. There are a lot of wastes generated every year from agriculture and forestry production, municipal waste or organic industrial. Laos has high potential of energy crops, which can be used as feedstock for biofuel production and energy used.
In the country is mainly in form of traditional fuels. Fossil fuels play important roles in energy sectors and global economy (Ullah K, 2015) and basic energy used by humans comes from fossil fuels. Fossil fuels not only dominate in field of energy but also involved in accumulation of greenhouse gases in atmosphere resulting in global warming due to which such fuels become widely organized as unsustainable (Bai F, 2014). Day by day the voice is raised universally against global warming due to change in the weather patterns, world‘s sea level increase enveloping lowland, deltas and islands and a tendency for global temperature increase.
Biologically produced fuels are looked upon with much interest and identified as potential alternative energy source (Parmar A, 2011). Renewable energy or alternative energy is a term used for an energy source that is an alternative to using fossil fuels. These alternatives are intended to address concerns about such fossil fuels has high carbon dioxide emissions, an important factor in global warming. Fossil fuels are non-renewable.
They are limited in supply and will one day be depleted (Zehner, 2012). 1 One of the most renewable energy is from bioethanol because bioethanol can be produced from agricultural feed stocks. It can be made from very common crops such as hemp, sugarcane, potato, cassava and corn. There has been considerable debate about how useful bioethanol is in replacing gasoline.
Concerns about its production and used relate to increase for food prices due to the large amount of arable land required for crops (The madness of biofuel, 2011), as well as the energy and pollution balance of the whole cycle of ethanol production. The main objective of this thesis is to successfully produce bioethanol from sugarcane bagasse. Sugarcane bagasse is an agricultural waste which was obtained from SRIWIJAYA laboratory in Palambang, Indonesia. The experiment used to produce bioethanol by using separate hydrolysis and can be made from glucose solution from fermentation process.
To get the result of ethanol concentration from sugarcane bagasse, the standardization of ethanol was used to determine the intensity of ethanol from each samples. The amount of ethanol was showed in peak area. Research’s objectives The objective of this research project is to: 1. Find out and describe ethanol production.
Find out the use of ethanol production from waste products that are lignocellulosic. To determine the optimum conditions of fermentation process for the production of bioethanol from lignocellulosic biomass. Producing (experiment) of ethanol from lignocellulosic biomass. Compering the economics of the production and yield of ethanol using separate hydrolysis and fermentation.
Research questions and hypotheses Bioethanol has a number of advantages over conventional fuels. It comes from renewable source or energy crops. To produce ethanol by not impact food consumption and feedstock the product can be development into the used of waste to produce ethanol fuel. There is ongoing research to answer the research questions.
Whether ethanol can be produced from sugarcane bagasse as raw material and yeast isolated from durian fruit as biological agent? b. How much bioethanol can be produced using Separately hydrolysis fermentation? c. How much energy goes into the ethanol production process and how much come out? 1. Limitations The limitation of the production of ethanol from cellulosic include: 1.
Pretreatment process to reduce lignin is needed 2. To apply all sugar content needs microbial agent which can ferment not only glucose but also xylose and arabinose. The need to find or genetically engineer organisms to efficiently ferment these sugars. Costs of collection and storage of low density biomass feedstock.
Definitions Bioethanol production process has three steps which are pretreatment, hydrolysis and fermentation process. The scope of this study was to determine the yield of bioethanol 3 can be produced glucose from sugarcane bagasse in fermentation process. The 150 ml fermentation will conducted to investigate the effect of temperature and fermentation time. The optimum ethanol production of glucose from sugarcane bagasse using yeast isolated from durian fruit was aimed.
Others parameter such as pH and the results will be compared the optimum ethanol from different hydrolysis and fermentation timed. Energy guide The need to meet the ever-increasing demand for energy is probably the greatest challenge that society has to grapple with in this new millennium. Virtually every aspect of life on planet Earth (heating, transportation, etc.) requires energy input in one form or another. Population growth has always been and will remain one of the key drivers of energy demand, along with economic and social development.
Growing population consume more energy, placed on arable land, the increase in the consumption of crude oil, cost of energy fuel are rising (Bartlett, 1994). Energy resources are the estimated maximum capacity for energy production given all available resources on Earth. They can be divided by type into fossil fuel, nuclear fuel and renewable resources. However, it has been recognised that global crude oil reserves are finite, and their depletion is occurring much faster than previously predicted (Grant, 2005; Möller, 2006; Bai et al.
In addition, shortterm price volatility has heightened apprehension about the future of global energy security (Hahn-Hägerdal et al. In 2008, before the global economic recession began, crude oil sold for over USD135 per barrel in the market. However, conventional petroleum is essentially non-renewable and intertwined with this practical impediment is an apparent moral dilemma of environmental pollution arising from its very usage (Wackett, 2008). The combustion of these hydrocarbons makes significant contributions to greenhouse gases (GHG) in the atmosphere and inevitably contributes significantly to global warming (Wigley, 2005).
The transport sector alone accounts for 60% of global oil consumption (International Energy Agency (IEA), 2008), 19% 5 of carbon dioxide and 70% of carbon monoxide emissions (Goldemberg, 2008). With the world human population projected by the United Nations to hit 9 billion and the number of cars 2 billion (World Business Council for Sustainable Development (WBCSD), 2004) by 2050, it is no longer sustainable to continue to combust fossil fuel without regard for the environment. Consequently, the need for environmentally sustainable and renewable energy sources cannot be overemphasized, given the rapid rate of global industrial development (Zaldivar et al., 2001; Gray et al. Bioethanol can be used in various blends with gasoline, such as 5% bioethanol (Demirbas, 2008), 10% & 20% (E10 & E20) (Gray et al., 2006), 22% (Wyman, 1994) or even 85% (E85) (Balat et al., 2008) and because of its favourable physicochemical properties, ethanol is considered an excellent alternative transportation fuel to gasoline that can considerably improve the quality of the atmosphere (Philippidis, 1993).
A major boost for the biofuels industry has come from automakers, GM, Chrysler and Ford, who have stated that half of all the cars they produce worldwide in 2010 will be ‗flex-fuel‘, or E85 (85% bioethanol)- compatible (Waltz, 2007). Other benefits come from using bioethanol as biofuel: it is totally biodegradable and sulphur free, and the products from its incomplete oxidation (acetic acid and acetaldehyde) are less toxic in comparison to other alcohols (Minteer, 2006). Renewable energy or alternative energy Renewable or alternative energy is a term used for an energy source that is an alternative to using fossil fuels. These alternatives are intended to address concerns about such fossil fuels, such as its high carbon dioxide emissions, an important factor in global warming.