International Journal of Scientific Research Engineering & Technology (IJSRET)

Volume 2 Issue 6 pp 362-365 September 2013 www.ijsret.org ISSN 2278 0882

Production of Bioethanol From Fruit Rinds by Saccharification and Fermentation

Suhas V Bhandari*, Arun Panchapakesan, Naveen Shankar, H G Ashok Kumar Department of Biotechnology, RV College of Engineering, Mysore Road, Bangalore - 560059

ABSTRACT
The generation of bio-fuels from wastes forms an attractive solution towards both waste management and energy generation. The most important roadblock faced in large scale application is the aeration during saccharification. The utility of fruit rinds as a possible source of cellulosic ethanol in a process without aeration was investigated by using rinds of four fruits namely Pineapple, Jackfruit, Watermelon and Muskmelon. The powdered rinds were subjected to saccharification by Trichoderma viride followed by fermentation with Saccharomyces cerevisiae. Significant amounts of reducing sugars were obtained at the end of the saccharification process, with jackfruit and pineapple rinds being the most effective at 10.28 mg.ml-1 and 10.18 mg.ml-1 respectively. The amount of ethanol produced after fermentation was analyzed by gas chromatography and found to be highest for the same fruits with yields of 4.64 g.l-1 and 4.38 g.l-1 respectively. The results indicate the promising future for generation of ethanol from cellulosic wastes on a large scale. Keywords Bioethanol, Fermentation, Fruit rinds, Trichoderma, Saccharification. India being an agro based economy generates nearly 350 million tonnes of wastefrom the vegetables, fruits and other organic materials [5]. Organic matter including fruit rinds is a major part of wastes generated daily by households, agricultural sector and food processing industries. Fruits are used on a small and large scale for household consumption and by food processing industries like pulp and jam manufacturers. In urban areas, a considerable portion of solid waste includes fruit waste generated by fruit juice vendors and restaurants. These industries and establishments usually discard the inedible parts of the fruits which include the exocarp commonly referred to as rind or peel. In most cases, these waste materials are dumped in landfills which lead unhygienic conditions. However, utilization of these waste materials in production of bio-fuels would be of great environmental and economic benefit as it could reduce the burden on conventional sources of energy and also get rid of the wastes. Ethanol an important biofuel, having high calorific value has the added advantage of being less polluting than most sources of energy that are in use today.Reports available suggest that previous natural substrates for ethanol production via saccharification have included sugarcane bagasse, wheat straw, corn, softwood etc. [69]. Fruit rinds are excellent sources of cellulose which can be used for the production of ethanol via saccharification followed by fermentation [10]. Most of the procedures followed in the laboratory are not feasible when the process is scaled up for application on a scale that would be useful to a large community, the most important parameter being the amount of power consumed just for aeration [11]. This study is thus aimed at processes which can be scaled up without great investments or tedious procedures. The work showcases a comparative study on utilization of the rinds of Pineapple, Watermelon, Jackfruit and Muskmelon by T. viride and fermentation of these sugars by S. cerevisiae.

I.

INTRODUCTION

In a world where the dumping of wastes is causing serious harm to the flora and fauna of the areas surrounding the dumping sites, the concept of using the wastes for production of energy forms a solution which is easily adoptable, cheap and efficient. One of the most abundant sources of energy in the world is the bio-polymer cellulose, which forms a major component of most plant and algal cell walls. The ability of organisms such as species of Trichoderma, Aspergillus, Clostridium, etc. to produce the enzyme cellulase enables them to hydrolyze this cellulose into its constituent glucose units [1-3]. The glucose can then be utilized by organisms of the genera Saccharomyces which can ferment the glucose into fuels such as ethanol [4].

IJSRET @ 2013

International Journal of Scientific Research Engineering & Technology (IJSRET)

Volume 2 Issue 6 pp 362-365 September 2013 www.ijsret.org ISSN 2278 0882

II.

MATERIALS AND METHODS

1. Microorganism and Culture media: T. viride and S. cerevisiae were procured from MTCC (Microbial Type Culture Collection, Chandigarh) Accession number 9699 and 170 respectively. T. viride was cultured on Potato Dextrose Agar (PDA) and S. cerevisiae on Yeast extract Peptone Dextrose (YPD) at 30C. The cultures were stored at 4C and subcultured every 30 days. 2. Processing of the substrates: The fruit rinds of Ananas comosus (Pineapple), Artocarpus heterophyllus (Jackfruit), Citrullus lanatus (Watermelon) and Cucumis melo (Muskmelon) were obtained from local shops and washed twice with distilled water before being prepared for use. The rinds were dried in a hot air oven at 65C for 24 hours and powdered using a grinder. 50 g of each rind was weighed and utilized as the substrate. 3. Saccharification: The processed fruit rinds were added to a broth containing yeast extract (5g/l) and peptone (10g/l). The media was autoclaved at 1210C and 15 psi pressure for 20 min and T. viride was inoculated under aseptic conditions. The flasks were then incubated at room temperature for a period of 144 hrs.

4. Reducing sugars assay: Reducing sugars assay was carried out according to the Dinitrosalicylic method [12]. Un-inoculated media was used as the control for the assay. The optical densities of the samples were measured against the blank at 540nm. The glucose concentration was then calculated using standard glucose curve. 5. Fermentation: After saccharification, the cultures were autoclaved and filtered using Whatman No 1 Filter Paper. The filtrate was then transferred into 250ml Erlenmeyer flasks, made airtight with cork and autoclaved. The flasks were then aseptically inoculated with 15mlovernight grown S. cerevisiae and incubated at room temperature for 96hrs. The ethanol produced was determined by Gas Chromatography using a NUCON Gas Chromatograph (5765 EPC) with a flame ionization detector. The carrier gas used was Nitrogen.

III.

RESULTS

Jackfruit and Pineapple rinds displayed similar reducing sugar concentrations over the period of the assay, terminating at 10.28 mg.ml-1 and 10.18mg.ml1 respectively after 144 hours, as can be observed in Fig. 1.

Figure 1 Amount of reducing sugars produced with different fruit rinds as substrates.

IJSRET @ 2013

International Journal of Scientific Research Engineering & Technology (IJSRET)

Volume 2 Issue 6 pp 362-365 September 2013 www.ijsret.org ISSN 2278 0882

Figure 2. Ethanol yield after fermentation of sugars produced using various substrates.

Watermelon and Muskmelon produced comparatively lower concentrations of reducing sugars with 4.16 mg.ml-1 and 7.43 mg.ml-1 respectively at 144 hours when the process was terminated by autoclaving. The amount of ethanol produced after 96 hours of fermentation with S. cerevisiae in various flasks is as shown in Fig. 2. Ethanol yield of jackfruit rinds was the maximum at 4.64 g.L-1 of the media followed by Pineapple, Muskmelon and Watermelon rinds with yields of 4.38 g.L-1, 3.08 g.L-1 and 1.89 g.L-1 respectively. The percentage conversion was calculated to be 84.2 %, 89%, 88.2% and 81.1% for Pineapple, Watermelon, Jackfruit and Muskmelon rinds respectively.

by use of this kind of a process is less. Hence, a balance between the two needs to be established for an economic process with sufficient yield. The amount of ethanol produced is slightly below that of Itelima et al, which can be explained by the same reasons, cited above. The ethanol yields can be further increased by using mutant strains of S. cerevisiae, as has been reported by Manikandan et al [15,16].

V.

CONCLUSION

This study has explored the possibility of using Pineapple, Jackfruit, Muskmelon and Watermelon fruit rinds for the purpose of bioethanol production. These fruit rinds usually end up in garbage dumps or biogas reactors. This study shows that there is potential for use of these fruit rinds in bioethanol production with minimal energy consumption to provide aeration. These cellulosic substrates usually have a lot of lignin content which prevents easy access for the microorganisms for saccharification. Thus some pretreatment methods like Acid or Base treatment is necessary to delignify these wastes and to obtain higher reducing sugar yields and hence higher ethanol yields also.

IV.

DISCUSSION

The amount of reducing sugars produced at the end of saccharification is significantly higher than that reported by Omojasola et al [9], which can be attributed to the different rinds and organisms used. Reducing sugar concentrations can be further increased by pre-treatment of the cellulosic wastes as has been reported by Tewari et al and Gomathi et al [13,14]. Nevertheless, on a large scale pre-treatment and agitation are tedious procedures and usage of a saccharification process without aeration forms an ideal solution. However, the comparative yield

IJSRET @ 2013

International Journal of Scientific Research Engineering & Technology (IJSRET)

Volume 2 Issue 6 pp 362-365 September 2013 www.ijsret.org ISSN 2278 0882

Also the use of engineered strains of T. viride capable of high cellulase activities will also help in increasing the final bioethanol yield.

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[12]Miller GL (1959). . Use of dinitrosalicylic acid reagent for determination of reducing sugar. Anal Chem. 31 (3): 426-428. [13]Tewari HK, Marwaha SS, Rupal K (1986). Ethanol from banana peels. Agric Waste. 16 (2): 135-146. [14]Gomathi D, Muthulakshmi C, Kumar DG, Ravikumar G, Kalaiselvi M, Uma C (2012). Production of bio-ethanol from pretreated agricultural byproduct using enzymatic hydrolysis and simultaneous saccharification. Mikrobiologiia. 81(2): 220-26. [15]Itelima J, Onwuliri F, Onwuliri E, Onyimba I, Oforji S (2013). Bio-ethanol production from banana, plantain and pineapple peels by simultaneous saccharification and fermentation process. Int J Envt Sci Dev. 4 (2): 213-216. [16]Manikandan K, Saravanan V, Viruthagiri T (2008). Kinetic studies on ethanol production using banana peel waste using mutant strain of Saccharomyces cerevisiae. Indian J Biotechnol. 7: 83-88.

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