Available online at www.sciencedirect.com ScienceDirect

Energy Procedia 65 ( 2015 ) 372 – 377

Conference and Exhibition - New, Renewable Energy and Energy Conservation (The 3rd Indo-EBTKE ConEx 2014)

Ethanol Synthesis From (Artocarpus heterophyllus Lam.) Stone Waste as Renewable Energy Source

Wahidin Nurianaa*, Wuryantorob

aDepartment of Mechanical Engineering,, Merdeka University Madiun, Indonesia bDepartment of Agronomy, Faculty of Agriculture, Merdeka University Madiun, Jl. Serayu, No. 79, Madiun 63133, East , Indonesia

Abstract

This study was aimed to develop renewable energy from jackfruit’s stone. The jackfruit’s stone was sliced and dried for 2 d to 4 d afterward the stoness were analyzed for fungi and microbes through microscope. The stone was retrieved from 9 mo and 1 yr shelf life through 2 d to 4 d drying process. The process was made in acid method resulted to high glucose on the solution level 25 %, pH 3.5, and temperature 135 oC in 3 min. The enzyme method was more simple and resulted 19.24 % higher glucose than acid method that is 17.36 %. Fermentation was using 0.03 g of Saccharomyces cerevisiae, 0.10 g of urea and 0.05 g of N-P-K on concentration solution of jackfruit’ flour by adding 0.3 mL to 0.7 mL of alpha–amylase, 0.2 mL to 0.6 mL of Gluco-amylase for and fermentation process 3 d to 6 d result of ethanol level is 11 % to 13 %.

©© 2015 2015 Published W. Nuriana, by Elsevier Wuryantoro. Ltd. This Published is an open by access Elsevier article Ltd. under the CC BY-NC-ND license (Peerhttp://creativecommons.org/licenses/by-nc-nd/4.0/-review under responsibility of the Scientific). Committee of EBTKE ConEx 2014. Peer-review under responsibility of the Scientific Committee of EBTKE ConEx 2014 Keywords: Artocarpus heterophyllus Lam. ; ethanol fermentation; hydrolysis; jackfruit stone; renewable energy.

Nomenclature

d day mo month yr year h hour min minute N nitrogen P phosphorous K potassium

* Corresponding author. Tel.: +62 812 5913 722; fax: +62 351 495 551. E-mail address: [email protected]

1876-6102 © 2015 Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Peer-review under responsibility of the Scientific Committee of EBTKE ConEx 2014 doi: 10.1016/j.egypro.2015.01.066 Wahidin Nuriana and Wuryantoro / Energy Procedia 65 ( 2015 ) 372 – 377 373

1. Introduction

The consumption of eco friendly energy source and renewable gasohol (a mixture of gasoline and ethanol) become a concerned due to the reduction of crude oil supply as the source of the main energy. Another trigger that increases the value of ethanol as fuel or fuel blending is the enactment of legislation reducing greenhouse specifically the Clean Air Act 1990 (in America and Japan) [1,2]. The focus of this research is to find alternative fuels using low cost and environmental friendly materials. Many countries appear to compete about developing the possible source energy. China, Taiwan, South Korea's success in developing alternative energy sources from cassava. Development of ethanol in Brazil has been running 30 yr with from sugar cane as raw material, and in the United States the development of ethanol from corn. Germany used wheat and corn fiber as renewable alternative energy sources, while Canada successfully in cultivating grass as an alternative energy source. Indonesia has developed the process production of ethanol from cassava and molasses [3-6]. Waste utilization from jackfruit stone become one of potential application as the raw material for ethanol production since it contains around 12.78 % of starch [7]. Jackfruit trees can be found easily in almost every part of Indonesia. Jackfruit commonly used as one of the ingredients in food processing especially for traditional food. Jackfruit stone also can be utilized as a snack and chips, but the utilization of jackfruit stone as other source of raw material for food processing still limited, and most of them will end up in the process production as waste. jackfruit’s stone dried for three days were contains 12.78 % of starch, 15.26 % of protein, 7.39 % of fat, 13.27 % of fiber, and 9.68 % of water. Even the jackfruit seed contains a significant amount of starch, however, it does not contain gelatin so for the utilization of this starch as raw material for cakes or snack production this starch should be mixed with wheat flour or starch and or starch [7-9]. Since the starch content in jackfruit stone is quite high and cannot be utilized directly as the main ingredients for food processing as well as a waste of magnitude + 40 % of jackfruit. But, in this study jacfruit’s stone can be utilized as raw material for ethanol production. Currently, study of jackfruit utilization especially it seed as renewable source for ethanol production is not much. Therefore, the use of jackfruit stone as feed stock for ethanol production is needed. The production of ethanol from jackfruit stone was carried out through acid hydrolysis and fermentation processes.

2. Material and method

2.1. Material

Jackfruit stone was sliced and dried in the sun ± 3 d until the moisture content of 15.38 %, it was sieved (use 80 mesh and 100 mesh sized sifter). Aluminium foil is a packing material when it burned in the furnace. Enzym Alpha- amylase, Gluco-amylase, Saccharomyces cerevisiae was obtained from the microbiology laboratory Bogor Agricultural University, Indonesia.

2.2. Method

The research was conducted in the microbiology laboratory. The jackfruit stone was cleaned and washed in distilled water. After soaking for 24 h, the outer and epidermis skin of the jackfruit stone were removed and subsequently washed with distilled water and drained. The clean jackfruit stone then sliced and dried under the sun for 2 d, 3 d, and 4 d The dry jackfruit seed then milled and sieved at certain particle size. The water content in the starch was analyzed by gravimetric method. Some part of the starch was stored in order to analyze the shelf life. The microbial and fungal growth in the starch was also observed under a microscope. A solution of jackfruit stone flour with concentration of 25 % was made by adding water into a known amount of starch. The mixture then liquefied at 90 oC by adding different amount of alpha-amylase enzyme (0.3 mL, 0.4 mL, 0.5 mL, 0.6 mL and 0.7 mL) for 20 min to 30 min. During the process, the mixture was under constant stirring, and the pH of the mixture was maintained at pH 4 to pH 6. After the liquefaction process had completed, the saccharification process was carried out, and the temperature of the mixture brought to 55 oC, and Gluco-amylase enzyme added to the system. After 3 h, the mixture was cooled until its temperature drop below 35 oC, and the mixture was separated using filtering system, the filtrate was collected in a beaker glass and the glucose content was 374 Wahidin Nuriana and Wuryantoro / Energy Procedia 65 ( 2015 ) 372 – 377

analyzed using SNI 01-2891-1992 method. The fermentation process was carried out by adding 0.03 g of Saccharomyces cerevisiae and other nutrients such as 0.10 g of urea and 0.05 g of N-P-K. The alcohol level was measured at each variation of the addition of Alpha- amylase and Gluco-amylase. A mixture of jackfruit stone starch and water (0.25 %) was hydrolyzed with hydrochloric acid as catalyst. During the process, the pH of the mixture was set at different pH (2.0, 2.5, 3.0, and 4.0). The temperature of the mixture was maintained at 135 oC for 30 min. After the hydrolysis process completed, the glucose content in the solution was analyzed by SNI 01-2891-199 method [5,8,10]. Ethanol synthesis from jackfruit stone elucidated in Fig. 1.

Jackfruit stone waste are feeled, sliced and dried (mantaining the availability of raw materials)

Made into starch Analyze : water (blend and sieve well) content, resistance

Hydrolysis process is conducted using HCl in Hydrolysis process is conducted using temperature 135 oC and stirred for 45 min with pH Alpha- amylase enzyme in temperature 90 oC for 2; 2,5; 3; 3,5; 4 conducted using hydrolisis process 30 min, sacharification process conducted for 2 h

Glucose test result (from enzyme process)

Fermented with saccharomyces cereviseae enzyme, urea, N-P-K

Beer: sediment + water + Analyze ethanol level ethanol

Separation and filtration Sediment

filtrat

Ethanol level test

Fig. 1. Ethanol synthesis from jackfruit seed.

3. Results and discussion

The shelf life of jackfruit stone as the raw material for glucose synthesis. The synthesis of glucose conducted by the acid method with solution concentration of 25 %, temperatures of 135 oC to 150 oC for 40 min at pH 2.0, 2.5, 3.0, 3.5, 4.0. It can be seen that the glucose content obtained from the acid method is smaller than the enzyme method, while the volume ratio of acid method is larger than the enzyme method Fig. 2 to Fig. 6 depict the glucose content in the solution obtained by two different methods, i.e. acid and enzyme methods.

Wahidin Nuriana and Wuryantoro / Energy Procedia 65 ( 2015 ) 372 – 377 375

Mean glucose

level (%)

Alpha-amylase addition (mL)

Fig. 2. Mean glucose level vs addition of Alpha-amylase (mL) by hydrolisis enzyme method

Mean glucose volume (%)

Alpha-amylase addition (mL)

Fig. 3. Mean of glucose volume ratio vs addition of Alpha-amylase by hydrolisis enzyme method

Mean glucose level (%)

PH after adding the HCl solution

Fig. 4. Mean glucose level vs pH by hidrolisis of acid method

376 Wahidin Nuriana and Wuryantoro / Energy Procedia 65 ( 2015 ) 372 – 377

Mean glucose volume (%)

pH

Fig. 5. Mean glucose volume ratio vs pH on the synthesis of acid method

The fermentation of glucose obtained from jackfruit stone flour with the addition of 0.5 mL Alpha-amylase produced higher content of alcohol. The increase of enzyme concentration decreases the alcohol content.

Mean ethanol level (%) et

Alpha-amylase (mL)

Fig. 6. Mean ethanol level at fermentation process

Jackfruit stone flour could be stored for 8 mo (sun dried for 2 d, 3 d, and 4 d). This flour should be put in dry storage, and can be used as raw material for large scale ethanol production for continuous process because of its sufficient availability. At a longer storage time (9 mo), the microorganism began to grow on the flour. The flour obtained from 4 d sun dried stone had a longer storage life due to less water content in the flour. Dried jackfruit stone was more resistance to microorganism than the powder form, and it could be stored for 1 yr without any grow of microorganism on the surface of the stone due to its harder physical characteristic. From this study the best operating condition for glucose synthesis from jackfruit stone flour using the enzyme method was obtained. The method used in this study referred to the synthesis of glucose from cassava starch and fresh cassava [9,10]. Sugar content obtained from enzyme method was 19.43 % with the volume ratio of glucose was 33.15 %. The synthesis of glucose using acid method produced glucose content around 17.36 % at pH 3.5 with the volume ratio of glucose around 43.23 %. The yield of dry starch to glucose level was 77.72 %. From cassava raw material, (15 to 25) % yield of cassava starch was obtained and from dry starch produce (80 to 95) % yield of glucose syrup [8]. In the saccharification process, starch solution change into glucose using acid catalyst (HCl) performed by the operating conditions based on the glucose synthesis from sago ( sagu Rottb.) and ganyong (Canna discolour L. syn. C. edulis) [4,10-12]. Glucose levels obtained from the synthesis of the enzyme method was greater (19.43 %) than glucose level that obtained from a synthesis of acid method (17.36 %). The addition of enzyme Wahidin Nuriana and Wuryantoro / Energy Procedia 65 ( 2015 ) 372 – 377 377

(Alpha-amylase) gradually break down the polysaccharides into disaccharides, and the saccharification process breaks down the disaccharides into monosaccharide (glucose). The volume ratio in the glucose synthesis using the enzyme method was smaller than using acid method. From the methods employed in this study, the synthesis of glucose using enzyme method produced a higher glucose yield than the acid method.

4. Conclusion Research study resulted that the utilization of jackfruit seed as the raw material for ethanol production contains carbohydrates/ in amount 12.79 %. The jackfruit stone as raw material for ethanol production can be used from whole kernel or its flour which had a resistance to microorganism in the storage for 1 yr for the whole kernel or 8 mo for the flour. Hydrolysis process using acid method (HCl) produced high glucose result at the following condition: (i) concentration of the starch in mixture was 25 %, pH 3.5, temperature was 135 °C, and processing time in 30 min, (ii) The enzyme synthesis method produced glucose in levels 19.43 % and 33.15 % respectively. The synthesis process by acid method produced glucose levels of 17.36 % at pH 3.5, the ethanol level of the enzymatic process and fermentation is 11 % to 13.4 %.

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