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  See discussions, stats, and author profiles for this publication at: Alcohol Production from Fruit and Vegetable Waste Article   in   International Journal of Applied Engineering Research · December 2013 CITATION 1 READS 9,507 1 author:Some of the authors of this publication are also working on these related projects: production of xylanase   View projectGirisha MalhotraManav Rachna Educational Institutions 4   PUBLICATIONS   3   CITATIONS   SEE PROFILE All content following this page was uploaded by Girisha Malhotra on 28 November 2016.  The user has requested enhancement of the downloaded file.  International Journal of Applied Engineering Research. ISSN 0973-4562, Volume 8, Number 15 (2013) pp. 1749-1756 © Research India Publications Alcohol Production from Fruit and Vegetable Waste Shilpa C., Girisha Malhotra and Chanchal  Department of Biotechnology, Manav Rachna International University, Sector - 43, Suraj Kund Badkhal Road, Faridabad, Haryana, India Abstract Fossil fuels create a negative impact on our environment as green-house gas emissions are harmful. Production of ethanol (as an alternative fuel) from food and agricultural waste was done in this research by bio-processing. Wastes from fruits, such as banana, orange, pineapple and pea peels were subjected to simultaneous saccharification and fermentation for 7 days by co-culture of Aspergillus niger and Saccharomyces cerevisiae. The ethanol yield was determined at 24 hours interval. The results of the study showed that after 7 days of fermentation, pineapple peels had the highest  biomass yield, followed by banana peels, orange peels, pea peels. The optimal ethanol yields were 8.34% v/v, 7.45 % v/v, 3.98 % v/v and 2.58 % v/v for pineapple, banana, orange and pea peels respectively. These indicate that pineapple and banana peels ethanol yields were significantly higher than orange and pea peel ethanol yield. Keywords : Saccharomyces cerevisiae, Aspergillus niger, ethanol  production, fruits waste, fermentation, sachcharification. 1.   Introduction Due to rapid exhaustion of fossil fuels there is an urgent need to resort to alternative fuels e.g. ethanol. The first large scale use of ethanol as a fuel happened during the early 1900s when petroleum supplies in Europe were short. Though ethanol is conventionally produced from petroleum by-products, bio ethanol can alternatively be  produced by fermentation technology using renewable raw material. Saccharomyces cerevisiae  is the most popular organism used for ethanol  production due to its high ethanol yield and high tolerance. Now a days, crops are the main source used for ethanol production. To achieve significant economic and   Girisha Malhotra et al 1750environmental benefit large amount of food wastes can be utilized to produce ethanol. Utilization of fruit waste for bioethanol production is one of the best options. One example of raw material is pineapple waste that is converted to bioethanol (Hossain et al., 2008). The wastes contain valuable components such as sucrose, glucose, fructose and other nutrients (Sasaki et al. 1991). Lignocellulose is the major structural component of woody plants and non woody plants. The use of mango peel as a source of pectin and fibre production also has been reported (Pandia et al., 2004). Grohmann et al. (1994; 1995; 1996; 1998) previously reported ethanol production from orange peel. Ethanol production from banana (Manikandan et al., 2008) and pineapple peels (Ban-koffi and Han, 1990) were also investigated. Dried orange peels have a high content of pectin, cellulose and hemi cellulose, which make it suitable as fermentation substrate when hydrolyzed. Insoluble carbohydrates are present in the cell walls of the peels, particularly in the form of  pectin, cellulose and hemicellulose. 2.   Literature Review Ethanol was first prepared synthetically in 1826, through the independent effort of Henry Hennel in Britain and S.G in France. Michael Faraday prepared ethanol by the acid-catalysed hydration of ethylene in 1828, in a process similar to that used for industrial synthesis of ethanol today. Almost all ethanol is being produced by fermentations using S. Cerevisiae . Karsch et al. (1983) evaluated the potential of  Zymomonas mobilis and Saccharomyces cerevisiae for ethanol production from glucose both aerobically and anaerobically. S. cerevisiae commonly known as Baker’s yeast has the ability to ferment a sugar solution poorly supplied with oxygen, resulting in the formation of alcohol and carbon dioxide.    Zymomonas mobilis  degrades sugars to  pyruvate using the Entner-Doudoroff pathway. The pyruvate is then fermented to  produce ethanol and carbon dioxide   as the only products (Farombi and Britton, 1999). Also  A. niger  produces enzymes such as amylase, amyloglucosidase, cellulases, lactase, invertase and pectinases. Maximum saccharification was achieved by hydrolysing banana-waste cellulose with a cellulase enzyme from Trichoderma reesei QM 9414. A yield of 1·38% and 0·78% (v/v) and 44·5% and 61·1% ethanol (mg g − 1reducing sugars) was achieved from cellulose and acid hydrolysed (2·5% at 15 psi for 15 min) banana peels, respectively. Grohmann et al. (1998) observed that  E. Chrysanthemi EC16 contains the PET operon from  Zymomonas mobilis on the plasmid pLOI555 which increases the organism’s ethanol production and decreases the final concentration of co-products (Beall and Ingram 1993).  Escherichia chrysanthemi EC16 fermentations of sugar beet  pulp produced 1.97 % (w/v) ethanol, less than the 2.55 % (w/v) ethanol produced by  E. Coli KO11 on the same substrate. In a study by Jayant Mishra et al. (2012), ethanol  production from fruit peels of pineapple, orange and sweet lime was investigated. Total amount of sugar in pineapple, sweet lime and orange was 0.5, 1 and 0.8% respectively. In the solid state fermentation, pineapple agro residue gives a maximum yield around 2.16% with yeast. With a change of strain to C. albicans , pineapple still   Alcohol Production from Fruit and Vegetable Waste   1751   gives a high yield of 1.08% for group A in 50 ml capacity. Pineapple gives a maximum yield of 1.87% with S. cerevisae . Lavarack B. P. et al. (2002) tried dilute acid hydrolysis of bagasse for conversion of hemicellulose to xylose, glucose, arabinose, acid soluble lignin and furfural. 3.   Materials and Methods 3.1 Routine Culture Maintenance Culture of S.cereviacae was maintained on YEPDA (1% yeast extract, 2% peptone,2% agar) slant stored at 4°C. The growth of S.cereviacae  confirmed plate count methods (Yeast malt extract medium,at 28°C.incubation period 2-3 days).  Aspergillus niger was cultured on PDA at pH 6.5 and 28  C. PDA was prepared using 20g dextrose, 20g agar and 4g potato extract dissolved completely in 500 ml water in a conical flask(pH-6.5). The mixture is then sterilized in an autoclave at 121  C for 15 minutes. (Kingsley Otulugbu, 2012 ) 3.2 Preparation of Pineapple, Banana, Orange and Pea Peels for Ethanol Production Pineapple, Banana, orange and pea peels were washed and their outer coats are removed, cut in small pieces and kept it in the sunlight for few days and then kept in oven for drying and stored in refrigerator prior to use. 3.3 Preparation of Growth Medium The growth medium prepared for ethanol production consists 20 g substrate (Pineapple, orange, banana or pea peel) in 250 ml of conical flask containing 100 ml of distilled water(pH-5.5). The flasks were autoclaved at 121°C for 20 minutes. This medium is poured in petriplates and set aside to solidify. 3.4 Preparation of Inoculum The cells of S .cereviacae were aseptically cultured in Yeast Extract Peptone Dextrose (YEPD) broth and incubated at 30°C for 24hrs. 3.5 Saccharification of the Fermentation Medium with  Aspergillus The substrate medium as prepared above was inoculated with spores of  Aspergillus. The culture was incubated at 28  C for 7days under rigorous aerobic culture. The samples were taken at regular intervals after every 24 hr for analysis. After 7 days in culture maximum total sugar was obtained. 3.6 Ethanol Production Medium (270 ml) was prepared and transferred to a Duran wide mouth bottle. The media was autoclaved at 121  C for 20 minutes and cooled. The culture broth (30 ml) from the saccharification step was added to the medium. This broth contains the cellulolytic enzyme complex elaborated by  Aspergillus . The bottle was inoculated with


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Jun 13, 2018
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