Bio-ethanol Production by Using Oil Palm Frond Juice (OPFJ)

Bio-ethanol output signal by Using oil Palm Frond Juice (OPFJ)OBJECTIVESTo enhance bioethanol out(p)put signal utilize OPFJ as subst priseTo optimized biotehanol production using various ferment elbow roomINTRODUCTION AND RESEARCH BACKGROUNDBrazil and USA is the first manufacturing businesss with the valet de chambre ethyl alcohol production nigh 5 carbon0 mills liters with an average of 73% of produces ethanol worldwide corresponds to fuel ethanol, 17% to beverage ethanol and 10% to industrial ethanol (Eufrozina NIGA, 2009). Bio-ethanol can be a product from an eatable arising which known as first generation bio-ethanol such as lucrecane and corns. Second generation of bio-ethanol is from lignocelluloses biomass is from non-edible source such as ornamentation crude biomass. This make hour generation of bio-ethanol is a better cream to replace fossil fuel without disturbing food sources. By converting the dispel into valuable product we can exituce the environsal p roblem. Lignocelluloses material nail about 50% 60% carbohyd evaluate in the form of cellulose and hemicelluloses which whitethorn be fermented to ethanol and 20%-35% of lignin (Mats Galbe et al, 2007). Nowa years typewriter ribbon rock petroleum color tree is one of the main source for the production of bio-ethanol but there be not much attention is been given to anoint laurel jade which are product that come from the tree component such as anele palm trunk and oil palm fronds.Malaysia is the worlds largest exporter of palm oil product. Malaysia produces a large amount of agro-industrial residue with oil palm industry itself produced about 33 million tons of residues per annum in the form of empty fruit bunch, fiber and shell (MPOB, 2009 Mohamed and Lee, 2006). Malaysia has produced about 51 million tons of oil palm fronds with 53% of the get palm biomass in year 2008 (Goh et al, 2010 MPOB, 2009). Bio-fuel that produces from palm oil tree is an environmental friendly therefore some interest are macrocosm shown to this sources. Mostly, the major(ip) parts of the solid biomass from the oil palm tree are being leave behind on the plantation is to be found as oil palm fronds.It has been reported that about 46837K tons of oil palm fronds has been produced in Malaysia in the year 2007 as an agri glossiness wastes. Palm oil frond is one of the useful raw materials for the production of bio-ethanol which is environmental friendly way. Pre-treatment of the oil palm frond need to be make to achieve a devout bio-ethanol production. However, ethanol production from lignocellulosic biomass is relatively expensive because of the latest technologies. The main defend are low yield and cost of the hydrolysis process (Sun and Cheng, 2002).Thus, oil palm frond juice is being introduced as another source for the production of bio-ethanol. old study has been shown that oil palm juice is suitable to used as turmoil providestock because there was no inhibitio n on microbial increment or product formation, there were no impurities, it was easy to be operated, and it had no risk on health and safety (Zahari et al, 2012). fossil oil palm frond expel has been proved by Zahari et al. (2012) contains high sugar content which is 76.09 2.85 g/l. there has been reported by Eze and Ogan (1988) that sucrose is the do hourant sugar in the oil palm frond sap that consist of 10% w/v, as for glucose and fruit sugar consist of et al, 2010. However in Malaysia, a study shows that glucose is the dominant sugar in the oil palm sap (Kosugi et al., 2010). Since there are many studies show that oil palm juice contains higher sugar content, a high level of the production of bio-ethanol from oil palm sap is come along study.A study by Nwachukwu (2008) shows that improving yeast resistance by protoplast fusion increased yields of ethanol by 16% v/v. Apart from that, types of agitation also one of the factor that improve the production of bio-ethanol. Thus, this project aim is to increase the yield of the bio-ethanol production using oil palm frond juice by various type of fermentation process.occupation STATEMENTUses of natural sources like petroleum is been used over the decades and the drive of this sources are being increases from time to time. Over the century, there are an increase of energy consumption with the increasing of world population, thus more exploitation countries become industrialized which there are possibility that the sources testament be depleted one day. Apart from that, petroleum is harmful to human and environment and with the increasing of fossil fuel ordain lead to increasing of vitamin C dioxide that eventually leads to global warming issue. Thus, an alternative sources of the fuels is been quest to overcome the precipitance of the fossil fuels. Bio-ethanol is one of the solutions to this problem. This is because bio-ethanol is a form of renewable energy source which are easy available, low cost and most efficient bio-fuel. Bio-ethanol offers a great advantages because it can be produced from various feedstock such as corn, sugarcane, red seaweed part sugar beet and many more (Mohd Dinie et al, 2013).Apart from that , bio-ethanol help in reducing air pollution and century dioxide accumulation.Nowadays, high demands of fuel are been constantly increase over the world. Brazil is been known as the largest producer of the sugarcane and a most competitive producer of the bio-ethanol in the world. The arises of bio-ethanol from sugarcane is a prove that energy sources are being run out and constant increasing of fuel cars around the world. Fossil energy gives an adjoin to the atmosphere because the burning of the petroleum result in increasing of carbon dioxide emission to the environment which is the main causes of greenhouse effect. product of bio-ethanol is increase significantly because many countries are looking for reducing oil imports, increasing plain economies and for be tter air quality (Eufrozina NIGA, 2009).4.0METHOD4.1Raw material homework and Juice Clarification. (Potential Utilization of vingt-et-un from Oil Palm (Elaeis guineensis) for Lactic blistering Production by Lactobacillus casei by S. Chooklin et al, 2011)Oil palm frond (OPF) go forth be harvest from oil palm tree from a local oil palm plantation.The leaves from the frond leave alone be cut off and discard.The harvest OPF will be press using sugar-cane machine press as in brief as possible after the frond was harvested.OPF juice produced will be bring in and filtered using coffee filter.The filtered OPF juice will be centrifuged at 10,000 revolutions per minute for 15 minutes.Supernatant will be stored at -20C out front use.4.2 OPF Juice carbohydrate Composition ( neutral spirits Production Using Immobilized genus genus genus Saccharomyces cerevisiae in Lyophilized Cellulose Gel by Eleonora Winkelhausen et al, May 2010)HPLC typeHPLC (Shimadzu lacquer ), provide with refract ive index DetectorType of pillar APS-Hypersil column ( diam of 250mm x 46mm)Mobile phase70% Acetonitrile and 30% de-ionized water supply column temperature40C with maximum operating temperature of 80C blend rate 0.6 ml/minSample volume 20lPressure 10MPa4.3Pure Culture Establishment (Study on Bioethanol Production from Oil Palm Trunk (Opt) Sap by Using Saccharomyces Cerevisiae Kyokai No.7 by Nina Farhana 2010) (Isolation of Microorganism from Oil Palm Sap by Nurul Nadia Ummira, 2011)100mL of the nutritious pedigree Medium supplemented with g/L Yeast Extract, 5 Peptone, 10 and 15% of glucose solution, 50 (Jamaludin, 2010) will be measured in a 250mL conic flask and will be autoclaved at 121 C in 15 minutes.Saccharomyces cerevisiae will be subculture into the alimentary Broth.The mixture will be incubated for 24 seconds at 150 revolutions per minute, 30oC.The suspension obtained will be subculture on the Nutrient Agar flip and will be incubated for another 2-3 days.The Nutrien t Agar slants will be stored in refrigerator at 4oC.4.4 Inoculums homework (Study On Bio ethanol Production From Oil Palm Trunk (OPT) Sap by Using Saccharomyces Cerevisiae Kyokai No.7 by Nina Farhana 2010) (Optimization study of ethanol fermentation from oil palm trunk, rubber wood and mixed hardwood hydrolysates using Saccharomyces cerevisiae by K.L. Chin et al , 2010)About 3 4 twine of pure culture will be transferred from agar slant into 100 mL of sterile Nutrient Broth in 250 ml conical flask plug with sterile cotton.The flask will be incubated for 24 hours at 30C and 150 rpm until reach measurement initial tightfistedness. The cell concentration will be standardized to 0.2 0.4 g/L (OD = 4.2) determined by turbidimetry at 600 nm.4.5 turmoila) hatful fermentation will be performed for the optimization of bio-ethanol production.b) 20% (v/v) of inoculums size will be inoculated into the bioreactor with the condition of pH 4.5 and temperature of 30C , air flow rate of 1 vvm , agitation of 200 rpm with the initial 02 concentration of 30 %.c) The fermentation process will be carried out for 42 hour and sample will be taken out every 3 hour and dissect for bio-ethanol production, sugar and cell biomass.d) All experiment will be run in triplicate.4.5.1 grasp fermentation.4.5.1.1Effect of sugar concentration (Effect of Cultural Conditions on grain alcohol Production by Locally free Saccharomyces Cerevisiae Bio-07 by Arifa Tahir et al, 2010)OPF juice volume use will be varied in the range of 30% to 50%.(30%, 40%, 50%)4.5.1.2Effect of agitation speed (Optimization of Fermentation Medium for the Production of neutral spirits from Jaggery Using Box-Behnken Design by Mary Anupama.P et al 2010)The fermentation will be carried out at different agitation rate in the range of 125 rpm to 175 rpm.(125, 150, 175) 4.5.2Fed Batch fermentation4.5.2.1Effect of feeding rate (Production of Ethanol by Fed- Batch Fermentation by Ngoh Gek Cheng et al, 2009)The fed batch ferm entation will be run match to the best optimized condition from the batch fermentation condition.substratum will be feed continuously into the bioreactor using peristaltic pump at glucose concentration (glucose concentration from OPFJ?) varying from 2 ml to 8 ml at feeding rate of 2 ml/h .(2 ml , 4 ml, 8 ml)4.5.2.2Effect of feed time (Production of Ethanol by Fed- Batch Fermentation by Ngoh Gek Cheng et al, 2009)a) Substrate will be feed from the interval of every one hour to three hour at feeding rate of 2 ml/h.( 1 hour , 2 hour , 3 hour )4.6Harvesting (Study on Bioethanol Production from Oil Palm Trunk (Opt) Sap by Using Saccharomyces Cerevisiae Kyokai No.7 by Nina Farhana, 2010)Each batch of fermentation will be terminated according to optimized time period.b) Sample (15 ml) will be taken out and the samples will be kept in refrigerator at4oC before analyze for bio-ethanol production, sugar and cell biomass.4.7Yeast strain and it improvement (High-level Production of Ethanol dur ing Fed Batch Ethanol Fermentation with a Controlled Aeration Rate and Non-Sterile Glucose demolish Feeding of Saccharomyces cerevisiae by Hyen-beom-seo et al, 2009)a) Saccharomyces cerevisiae will be produce on Nutrient Broth (yeast extracts, 10g/l peptone, 20g/l glucose, 20g/l and agar, 20g/l) slant in a 30 ml universal bottle.b) The slant culture will be exposed with UV light (6 watt, 254nm) for 15 second interval at a distance of approximately 7 cm from the slant.c) A loopful of irradiated slant culture will be streak onto a Nutrient Broth agar plate and subsequently incubated for 3 days at 27C.d) After that the colonies will be incubated in Nutrient Broth medium containing 100 g/l of ethanol at 27C while shaking at 100 rpm for 5 days to select resistance colonies.4.8 Analysis method4.8.1 Sugar content by HPLC (Oil Palm Fronds Juice as Future Fermentation substrate A Feasibility Study by Che Mohd Hakiman Che Maail et al, 2014)HPLC typeHPLC (Shimadzu Japan ), equipped with refr active index DetectorType of column APS-Hypersil column ( diameter of 250mm x 46mm)Mobile phase70% Acetonitrile and 30% de-ionized waterColumn temperature40C with maximum operating temperature of 80CFlow rate 0.6 ml/minSample volume 20lPressure 10MPa4.8.2Cell dry determination (Study on Bioethanol Production from Oil Palm Trunk (Opt) Sap by Using Saccharomyces Cerevisiae Kyokai No.7 by Nina Farhana 2010)Cell suspension will be vaccum-filtered using 0.45m filter paperCell suspension for each samples will contribute to be rinsed 2x with distilled water.Each sample will be dried at 70C more than 24 hours until constant weight achieved. desiccate filter paper will be weighed on a analytic balance to measure the biomass.4.8.3 Ethanol concentration using GC (Production of Ethanol by Fed Batch Fermentation by Ngoh Gek Cheng et al, 2009)Column RT-Q-BOND (inner diameter of 0.32 mm)Carrier gas Helium gasDetector Flame ionisation detector (FID)Temperature 200 CFlow rate 21.9 mL/minPressur e 71.1 kPaHolding time 5 minutes% bioethanol yield is calculated using this formula5.9 OPTIMIZATION OF ETHANOL FERMENTATIONRSM analysis will be conducted using Design Expert Software.The optimized parameters will be substantiate by running the new fermentation as suggested.6.0 ANALYSISThe implication of difference between each test variable will be determined using one way ANOVA analysis and to the lowest degree Significance Test, computed using SPSS version 21.0 software. All tests will be done with a confidence interval of 95%.7.0REFERENCESNgoh Gek Cheng, M. H., Andri Chahyo Kumoro, Tham, C. F. L. A. M. 2009. Production of Ethanol by Fed-Batch Fermentation. Pertanika J. Sci. Technol., 17, 399 408.Hyeon-Beom Seo, Seung Seop Kim,Hyoen0Yong Lee and Kyung Hwan Jung (2009).High level Production of Ethanol during Fed-Batch Ethanol Fermentation with a controlled Aeration Rate and Non-Sterile Glucose Powder Feeding of Saccharomyces cerevisie .Biotechnology and Bioprocess Engineer ing,14, 591-598.Jamaludin, N. F. M. 2010. Study On Bioethanol Production From Oil Palm Trunk (Opt) Sap By Using Saccharomyces Cerevisiae Kyokai No.7(ATCC 26422). Bachelor of Chemical Engineering (Biotechnology), Universiti Malaysia Pahang.Zahari, M. A., Zakaria, M. R., Ariffin, H., Mokhtar, M. N., Salihon, J., Shirai, Y. Hassan, M. A. 2012. Renewable sugars from oil palm frond juice as an alternative novel fermentation feedstock for value-added products. Bioresour Technol, 110, 566-71.Che Mohd Hakiman Che Maail, Hidayah Ariffin,Mohd Ali Hassan,Umi Kalsom Shah and Yoshihito Shirai (2014).Oil Palm Frond Juice as Future Fermentation Substrate A Feasibility Study.Bioresource Technology,110,566-71.Miller, G.L., Use of dinitrosalicylic corrosive reagent for determination of reducing sugar,Anal. Chem.,31,426, 1959.GANTT CHART AND MILESTONES