IJPAS International Journal of Pharmaceutical and Applied Sciences/2 (1)/2011 ICV (Impact Factor 2010): 2.92 ISSN 0976-6936 20 Study And Isolation Of Tannase Enzyme Production Bacteria From Tea Waste Dump Soil Site. A. Dave 1 , H. A. Modi 2 , N. Chavada 3* 1. Department of life science, Gujarat University, Gujarat-380009 2. Department of life science, Gujarat University, Gujarat-380009 3. Institute of Agribusiness management, Navasari Agriculture University,Navasari-396450, Gujarat * Corresponding author. Tel: +91-9724927027;E- mail:chavadanikul@yahoo.in Abstract Tannase (Tannin acylhydrolase EC.3.1.1.20) is an enzyme which catalyses the hydrolysis of tannic acid. It also acts on esters of gallicacid commonly found in tea extracts. The hydrolysis of these esters enhances the solubility of tea in cold water, in present work screening of tannase production bacteria from tea waste dump site and identification of hydrolytic tannase producing organisms. Production of crude extra cellular hydrolytic tannase from the bacterial isolate by submerged fermentation and also study optimization of factor affecting tannase activity. It is based on the spectrophotometric determination of gallic acid formed by the enzymatic hydrolysis of tannic acid. The kinetic parameters of the enzymatic reaction have been studied and an assay procedure has been formulated Keywords: Tannase enzyme producer, submerged fermentation, tea dump soil waste. 1. Introduction Hydrolysable tannins such as gallotannin and ellagitannin are widely distributed in the plant kingdom. these tannins bind readily with proteins and other macromolecules to form indigestible or in soluble complexes, thereby not only reducing nutritional value of animal feed (1) Tannase (tannin acylhydrolase)break specifically the ester bonds to diminis the tannins’ binding property.it has long been known that several fungi species such as aspergillus spp. and penicillin spp. are capable of producing a larger amount of this enzyme For industrial use, the enzyme is invariably of fungal origin (2) Over the past decade ,many bacterial species have also been reported to produce tannase, the include streptococcus gallolyticus (3) .Bacillus lichenifomis and several lactobacilli species (4) .in the course of further characterizing tannase-producing bacteria ,we have noticed appreciable variation in size of the clear zone among the isolates grown on tannin treated plate (3)-(4) .Tannase or tannin acyl hydrolase catalyses the hydrolysis of the ester bonds present in the hydrolysable tannin and gallic acid ester ,the main commercial application of tannase are in the preparation of instant tea and manufacture acron liquor. Tannase is also used for the production of gallic acid, which is an impartment intermediately compound in the synthesis of anti bacterial drugs. Beside tannase is used as clarifying agent in some wines, fruit juices and coffee flavored soft drinks. Tannase is an extracellular, inducible enzyme produced in the presence of tannic acid by fungi, bacteria and yeast (5) .The first step in the development of microbial enzyme production is the lineage selection. Extra cellular enzymes were preferred because they are easily extracted and do not require expensive extraction methods. Studies on the production of tannase using solid, liquid and submersed fermentation have been reported ductionprocesses (6)-(7) . the fermentation broth can use by-products such as wheat bran, rice bran, sugar beet pulp, fruit pulps, banana waste, cassava waste and coffee residues, adding tannic acid. The use of by products or residues rich in sources of carbon for fermentation purposes is an alternative way of solving pollution problems that can be caused by incorrect disposal in the environment In the present work isolation and screening of tannase producing bacteria, from waste tea dump site that check its activity at different pH and salt concentration .bacterial tannase produce by submerged fermentation process and verify it’s active by enzyme assay. 2. Materials & Methods 2. 1 Screening and isolation: Various tea waste dump site soil sample ware collected and enrichment culture method was used for tannase production. Enrichment culture was spread on tannin agar plate and screening was done after incubation at 37 C for 48 hours. Then after colonies were isolated (8) . Enrichment culture broth composition: tannin broth (in 100 ml).