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Glucose Production of Cellulose Hydrolysis Enzymatically on Rice Straw

Novia, Hermansyah dan Selvia Aprilyanti

Environmental Technologies of Magister Chemical Engineering, University of Sriwijaya

Email : noviasumardi@yahoo.com, selviaaprilyanti@yahoo.co.id

Rice straw is a by-product of agricultural activities resulting in large enough quantities in Indonesia. Rice straw has the potential to be used as raw material for alternative energy, namely bioethanol or other alternative fuels. Rice straw contains cellulose and hemicellulose that can be used to produce glucose by enzymatic hydrolysis. Enzymatic process is environmentally friendly process renewable raw materials. Enzymatic hydrolysis performed using cellulase enzymes to produce glucose. The purpose of this study was to determine the effect of hydrolysis time and enzyme concentration used for glucose production from rice straw that had been pretreated by alkali and ozonolisis. In this study, a 50 grams rice straw pretreated was dissolved in 100 ml of medium and the pH was adjusted to 5. The parameters varied are the hydrolysis time 5,10, 15, 20 and 25 hours, whereas the concentration of enzyme added was 10%, 20%, 30% and 40% w / v of the weight of rice straw. The results showed that the greatest glucose obtained was 0.218 g/L at 25 hours of hydrolysis and enzyme stater volume of 40% of the weight of rice straw. Keywords : enzymatic hydrolysis, glucose, rice straw

I. INTRODUCTION Rice straw is one of the largest agricultural wastes in southern Sumatra. Rice production per hectare can reach 10-15 tons of dry matter each harvest, depending on location and crop varieties. Each kilogram of rice straw yields results in the production of 1 - 1.5 kg of rice straw. Global production of rice straw around 650-975 million tons per year and is mostly used as animal feed and as agricultural waste (Binod et.al, 2009). Burning of rice straw is one of the practical ways to address waste rice straw, but this could increase air pollution and negative impacts on public health. Therefore, rice straw needs to be re-processed into useful materials but does not pose a problem for the environment. Rice straw containing lignin, cellulose, and hemicellulose. Cellulose content of rice straw is quite large, which is about 35-45%, so that rice straw can be used to produce glucose (Galleti and Antonetti., 2011). Presence of lignin in the straw will inhibit the

action of enzymes to convert cellulose into glucose, therefore it is necessary to pretreatment (pre-treatment) to reduce the amount of lignin in rice straw making cellulose more easily react with the enzyme. Conversion of cellulose to glucose is done with the help of cellulase enzymes. Cellulase enzymes will accelerate the hydrolysis of cellulose to glucose (Fatma et al, 2010). Pretreatment technologies are grouped into physical, chemical, physicochemical and biological. In this study, conducted physically Pre-treatment which aims to minimize the size of the biomass and reduce the crystallinity of cellulose, is done by cutting and grinding. Pre-treatment is then performed chemically by using a solution of NaOH / alkaline (Yoswathana et al, 2010; Roslam et al, 2011) and the ozonolysys to degrade lignin and hemicellulose in biomass (Cubero et al, 2009). Hydrolysis is the process of solving the polysaccharides in lignocellulosic biomass, namely cellulose and hemicellulose into

sugar monomers constituent. Perfect cellulose hydrolysis yield of glucose, while hemicellulose monomer sugars produced some pentose (C5) and hexose (C6). Hydrolysis can be done chemically (acid) or enzymatic. Enzymatic hydrolysis has several advantages over acid hydrolysis, among other things: no sugar degradation hydrolysis results, a milder process conditions (low temperature, pH neutral), has the potential to provide high yields, and equipment maintenance costs are relatively low because no corrosive materials (Taherzadeh and Karimi, 2007; Sun Y and Cheng, 2002). Enzymatic process is environmentally friendly process renewable raw materials (renewable raw materials). Currently, enzymatic hydrolysis is a very promising technology to convert biomass such as rice straw to glucose. Aspergillus Niger is a fungi that produce cellulase enzymes and can hydrolyze cellulose. Aspergillus niger to produce cellulase enzymes in rice straw media with high enzyme activity (Ong et al, 2012). This study aims to obtain the best operating conditions on enzymatic hydrolysis of rice straw to glucose by using Aspergillus Niger. II. RESEARCH METHODOLOGY 2.1 Preparation of Raw Materials Biomass in the form of rice straw obtained from paddy fields in the region MUSI 2 Palembang. Rice straw was destroyed using grinding machine (mill). Then dried in the hot sun for 10 days. Then heated in an oven at 45 C to remove the water content in the biomass. Once dry, the smaller the tool grinder to a certain size and then sieved to obtain the size of the straw 0.25 mm (80 mesh). 2. 2 Alkaline pre-treatment Rice straw that had been measured lignin content and moisture content of 50 grams into the bottle lid (lid erlenmeyer) and dissolved in 5% NaOH solution of 500 ml (comparison ratio (w / v) rice straw: NaOH = 1: 10). Further samples were incubated in a container in the

water bath at a temperature of 850C for 1 hour (Yoswathana et al, 2010). After 1 hour, samples were filtered and washed with distilled water until neutral pH. Further samples of solids dried in an oven at a temperature of 1050C to constant weight. lignin content of the final sample is calculated using the Kappa method. 2.3 Ozonolysys Pretreatment Ozonation process is carried out at a constant voltage, the 8500 Volt. Treatment of oxygen flow rate and the sample size of each variable at 5 L / min and the sample size 0.177 mm - 0.25 mm. Ozonolisis process rice straw held in ozonation reactor. 50 gram samples of rice straw in a state of 10% moisture content is inserted into the reactor ozonolisis. Before mengozonolysis biomass, ozone levels were analyzed first by iodometric method. 2% KI solution into a tube inserted into the analysis-1 (bottom tube) and analysis of the 2nd tube (tube top). Ozone gas flowed into the ozonation reactor for 15 minutes. After jetting 15 minutes, ozone is formed analyzed from KI solution to the tube-1 with the iodometric method. While the rest of the gas supplied to the 2nd tube analysis. The content of residual ozone ozonolisis reaction was analyzed as the tube analysis to-1 with the iodometric method. The samples were diozonasi, analyzed the content ligninnya using Kappa method. 2.4 Enzymatic hydrolysis The results of pre-treatment put into 500 ml Erlenmeyer then added 100 ml of distilled water and adjust pH 5. Then heated in an autoclave at 100 C for 30 minutes. Rice straw pulp allowed to cool. Cellulase enzyme is added according to treatment variables into mush the rice straw and Erlenmeyer sealed with a cork. Treatment of hydrolysis time, each consisting of the 5, 10,15, 20 and 25 hours, while the

treatment of the enzyme concentration is 10%, 20%, 30% and 40%. Enzyme concentration of 10% means that the addition of the enzyme with 10% of total enzyme fraction (5 ml enzyme per 50 grams of dry biomass) and close tightly with a cork erlenmeyer. Then placed on a rotary shaker at 160 rpm for 25 hours. Glucose hydrolysis results were analyzed by the method of luff schoorl.

glucose in each sample are shown in Figure 1

III. RESULTS AND DISCUSSION 3.1 Results Glucose analysis results obtained in this study of enzymatic hydrolysis process results shown in Table 1, ie Table 1. Results Analysis of Glucose 3.2 Discussion
a. Effect of Hydrolysis Time on Glucose

Figure 1. Effect of hydrolysis time on glucose Basically increasing hydrolysis time can increase glucose levels generated along with the many volumes of enzyme were added. This is because the longer the time given the more active side of the enzyme cellulase work cut carbon chains in the structure of cellulose into simpler structures such as glucose or a long time allowing the continuous reaction of the enzyme to hydrolyze cellulose to glucose. Vice versa, the less time given to carry out the process of hydrolysis eat the less cellulase enzyme works to hydrolyze cellulose to glucose, so glucose produced is also getting smaller. Seen from a glucose test results obtained as in Figure 1 above. Glucose test results did show a trend of increase in glucose values with increasing hydrolysis time. Of Figure 1 obtained the highest glucose value of 0.218 g / L at 25 hours hydrolysis time. b. Effect of Enzyme Concentration on Glucose Addition of enzyme concentration on the amount of the fixed substrate, generally increases the rate of glucose production (Figure 2).

In the study with the variation of the

Hydrolysis time 5hours Enzim Concentration 10% 20% 30% 40% 10 hours 10% 20% 30% 40% 15 hours 10% 20% 30% 40% 20 hours 10% 20% 30% 40% 25 hours 10% 20% 30% 40% Glucose (g/L) 0.0048 0.0168 0.024 0.036 0.0048 0.0745 0.072 0.117 0.117 0.122 0.129 0.147 0.198 0.206 0.211 0.216 0.206 0.211 0.213 0.218

weight of 50 grams with rice straw hydrolysis time variations used were 5, 10, 15, 20 and 25 hours, and the concentration of enzyme used 10%, 20%, 30% and 40% w / v, obtained content

0.25 0.2 Glucose (g/L) 0.15 0.1 0.05 0 0 20 40 Enzyme Concentration (%) 60

Acknowledgements Authors would like to thank the Higher Education research grant through the National Strategic hisbah 2012. REFERENCES Binod, P., Sindhu.R, Singhania, R.R, Vikram. S, Devi. L, Nagalakshmi. S., Kurien. N., Sukumaran, R K (2010), Bioethanol Production from Rice Straw, Bioresource Technology 101 : 4767- 4774 Cubero, M.T.G., Benito, G., Indacoechea, I.,Coca, M., Bolado, S (2009), Effect of Ozonolysis Pretreatment on Enzymatic Digestibility of Wheat and Rye Straw, Bioresource Technology 100 :pp.1608 1613. Fatma, H.A.E., dan Fadel, M (2010), Production of Bioethanol Via Enzymatic Saccharification of Rice Straw by Cellulase Produced by Tricodherma Reesei Under Solid State Fermentation, New York Science Journal : pp.72-78 Galleti, A.M R dan Antonetti, C. (2011), Biomass Pre-treatment : Separation of Cellulose, Hemicellulose and Lignin. Existing Technologies and Perspectives, Department of Chemistry and Industrial Chemistry, Universitas of Pisa, Italy. Ong, L G.A., Chan, C. H dan Chew, A.L (2012), Enzymatic Hydrolysis of Rice Straw : Process Optimization, Journal of Medical and Bioengineering (JOMB) Vol.1.pp.14-16. Roslam, A.M., Yee, P.L., Shah, U.K.M., Aziz, S.A dan Hassan, M.A (2011), Production of Bioethanol from Rice Straw Using Cellulase by Local Aspergillus sp., International Journal of Agricultural Research 6, pp :188193.

hydrolysis time 5 hr hydrolysis time 15 hr hydrolysis time 25 hr

Hydrolysis time 10 hr hydrolysis time 20 hr

Figure 2. Effect of enzyme concentration on glucose Can be seen in Figure 2, an increase in glucose content of the addition of the enzyme concentration of 10% to 40% addition of enzyme concentration. For example, the hydrolysis time 5 hours, an increase in glucose were almost linearly with increasing concentration of enzyme added. The increase in glucose levels along with the addition of large concentrations of cellulase enzymes is because many levels of cellulase enzymes resulted in the active site of enzymes to break down cellulose into glucose chains increase so that the enzyme activity increases. The greater activity of the enzyme, the more active side available enzyme to break down cellulose into glucose up. The best concentration of enzyme to hydrolyze rice straw is the addition of the enzyme concentration of 40%. Therefore, it can be concluded that the best 25-hour hydrolysis time and enzyme concentrations of 40% w / v glucose will be obtained at 0.218 g / L.

IV. CONCLUSION From the analysis of the Luff-Schoorl method can obtain the highest glucose was 0.218 g / L at 40% addition of enzyme concentration and hydrolysis time of 25 hours.

Taherzadeh, M.J dan Karimi, K.(2007), Acid-Based Hydrolysis Processes for Ethanol from Lignocellulosic Materials : A Review, BioResources 2(3), pp : 472499 Yoswathana, N., Phuriphipat, P., Treyawutthiwat, P. and Naghi E. M.(2010), Production Bioethanol from Rice Straw, Energy Research Journal 1 (1) pp.26 31.