Available online at www.scholarsresearchlibrary.com Scholars Research Library Archives of Applied Science Research, 2012, 4 (4):1763-1770 (http://scholarsresearchlibrary.com/archive.html) ISSN 0975-508X CODEN (USA) AASRC9 1763 Scholars Research Library Screening, isolation and production of lipase/esterase producing Bacillus sp. strain DVL2 and its potential evaluation in esterification and resolution reactions Davender Kumar 1a , Lalit Kumar 1 , Sushil Nagar 1 , Chand Raina 2 , Rajinder Parshad 2 , Vijay Kumar Gupta 1 * 1 Department of Biochemistry, Kurukshetra University, Kurukshetra-136119, India 2 Indian Institute of Integrative Medicine (IIIM), Canal Road, Jammu-Tawi (CSIR), India _____________________________________________________________________________________________ ABSTRACT The isolate DVL2 was isolated from common city garbage using the tributyrin as substrate. The isolate resulted in orange fluoresce under UV light on rohodamine olive oil agar plate detecting the lipase production. Three production media (PM1, PM2 and PM3) were evaluated for lipase/esterase production. In culture filterate (extracellular enzyme) and cell free extract i.e. extract after sonication of cells (intracellular enzyme), both lipase and esterase activity were detected. But esterase activity was found to be associated only with bacterial cells. The maximum intracellular (112 IU/L) and extracellular (33 IU/L) lipase production were obtained in Production medium 2 after 24 and 36 h respectively whereas the maximum production of esterase (extracellular, intracellular and membrane bound) was obtained in Production medium 2 after 24 h. The DVL2 lipase/esterase was found to esterify stearic acid with ethanol resulting in the formation of ethyl stearate which was confirmed by thin layer chromatography. Furthermore DVL2 lipase gave positive results when applied for resolution of chiral auxillary viz. 1-acetyl phenyl ethanol. Key-words: Bacillus sp., Lipase, Esterase, Esterification, Tributyrin Agar Plate Assay . _____________________________________________________________________________________________ INTRODUCTION Lipolytic enzymes [Esterase (E.C. 3.1.1.1) and Lipases (E.C. 3.1.1.3)] belong to a group of enzymes whose biological function is to catalyze the hydrolysis of triacylglycerols into diacylglycerols, monoacylglycerols, free fatty acids (FFA) and glycerol [1]. Many attempts were done by researchers to classify lipolytic and esterolytic enzymes [2]. Industrial applications may require specific enzymes like leather industry mostly require lipase and dairy industry require esterase [3]. Lipases and esterases can be distinguished on the basis of their substrate spectra, as esterases catalyze the hydrolysis of carboxylic ester bonds of short chain fatty acids (<10 carbon atoms) while true lipases have marked preference for long chain fatty acids (>10 carbon atoms) as substrates [4]. Lipase should be activated by the presence of an interface, that is, its activity should sharply increase as soon as the triglyceride substrate forms an emulsion. Lipoytic enzymes are highly diversified in their industrial application. They have emerged as key enzymes which find usage in food, dairy, paper, textile, leather and detergent industries, waste water treatment, production of fine chemicals, pharmaceuticals and cosmetics, synthesis of surfactants and polymers, vegetable fermentation and meat product curing [5-10]. They can catalyze esterification, interesterification, and transesterification reactions in non- aqueous media with high chemo-, regio- and/or enantioselectivity [11,12]. Due to numerous biotechnological