JOURNAL of APPLIED AND INDUSTRIAL BIOTECHNOLOGY in TROPICAL REGION, Vol. 2, No.1, Apr 2009 ISSN:1979-9748 (Special Edition) Immobilized Xylanase from Streptomyces sp. 45I-3in Corncob Xylan Hydrolysis 1 Immobilized Xylanase from Streptomyces sp. 45I-3 in Corncob Xylan Hydrolysis Anja Meryandini 1,2 , Titi Candra Sunarti 3 , Ferry Mutia 3 , Niken Financia Gusmawati 4 and Yulin Lestari 2 1 Research Center for Natural Resources and Biotechnology IPB, PAU Building, IPB Darmaga Campus 16680 E-mail : ameryandini@yahoo.com 2 Department of Biology, FMIPA-IPB, Animal Husbandry Building, 5th Fl Wing 1, IPB Darmaga Campus 16680 3 Department of Agriculture Industrial Technology, FATETA-IPB, IPB Darmaga Campus 16680 4 IPB Graduate School-Biotechnology, PAU Building, IPB Darmaga Campus 16680 Abstract Xylan extraction from corncob was done using alka- line as a solvent. Xylan extraction from corncob yielded 10.9%. One percent of corncob xylan was used as a substrate to produce the xylanase, as comparison, oatspelt xylan was used. Immobiliza- tion of xylanase was performed using 1% Eudragit TM S100 solution (w/v), with 5:1 volume ratio of xyla- nase. Activity of the immobilized xylanase de- creased to 23.97% compared with free xylanase. Immobilized xylanase had optimum pH of 6.0, and temperature at 40 o C, had also thermal stability at 30–40 o C for 1 h. Immobilized xylanase could be re- used, but its activity decreased to 52.38% after 3 times applications. Keywords : immobilization, xylanase, Streptomyces sp. 45I-3, Eudragit TM S100 INTRODUCTION Increased production in agriculture is followed by increasing in agricultural wastes. These wastes generate environmental problem. Generally, agricul- tural wastes still contain number of nutrients and can be converted to valuable product or used as growth medium for microorganism. Therefore, uti- lizing agricultural waste will overcome environmen- tal problem. Corncob is one of abundant agricultural wastes, because maize represents carbohydrate source after rice. Based on BPS data, maize production in 2004 reached 11.35 million ton, so that corncob waste was also abundance. Anonim (1982) reported that corncob contains cellulose (40%), hemicellulose (36%), lignin (16%) and other ingredients (8%). The main component of cellulose is xylan, which can be hydrolyzed by xylanase. Various kinds of bacteria, fungi and khamir are reported to be able to produce xylanase, and one of them is genus Streptomyces. Streptomyces sp. 45I-3 produces xylanase which have properties such as optimum activity at 50 o C and pH 5.0 (Meryandini et al., 2007). *Correspondence to: Anja Meryandini Tel: +62 21 ; Fax: +62 21 e-mail: ameryandini@yahoo.com Immobilized enzyme has some advantages com- pared to free enzyme. It can minimize enzyme cost, is easy to separate the enzyme from reaction solu- tion, can stop the reaction rapidly (or vice versa), can minimize effluent problem, can be reused and applied in continuous reaction, could be used in de- velopment of multienzyme system reaction, and in some cases, can improve enzyme stability (Shuler & Kargi, 2002). Hence, the aims of this research were to immobilize and characterize xylanase from Strep- tomyces sp. 45I-3 using Eudragit TM S100 matrix and also to use the immobilized xylanase to degrade corncob xylan. MATERIALS AND METHODS Materials: Prime material was corncob variety Bisma obtained from Agricultural Research Center of Biotechnology, Cimanggu, Bogor. Xylan extraction from corncob: Corncob pow- der was mixed with sodium hypochlorite (NaOCl) solution to begin delignification process, and xylan was extracted from corncob based on Anggarini (2003) using 15% NaOH. Determination of concentration of xylanase- inducing media: Streptomyces spp. isolate was inoculated to xylan containing media (1% yeast extract, 10.3% sucrose and 0.5% xylan) or 0.5:1 and 1.5% corncob xylan. Xylanase activity was determined with DNS method based on Miller (1959) with xylose as standard. Reducing sugar was determined by spectrofotometer at wave length of 540 nm. One unit of xylanase activity was defined as the amount of enzyme which produce 1 μmol xylose in 1 min. Immobilization of Streptomyces spp. 45I-3 xylanase: Crude extract of xylanase was immobilized with Eudragit TM S100 based on modified method of Sardar et al. (2000). Determination of Eudragit TM S100 solution concentration was conducted to find out the