© 2016 Maria Carolina de Albuquerque Wanderley et al. This is an open access article distributed under the terms of the Creative Commons Attribution License - NonCommercial-ShareAlike Unported License (http://creativecommons.org/licenses/by-nc-sa/3.0/ ). Journal of Applied Biology & Biotechnology Vol. 4 (04), pp. 001-010, July-August, 2016 Available online at http://www.jabonline.in DOI: 10.7324/JABB.2016.40401 Production and Characterization of Collagenase by Penicillium sp. UCP 1286 Isolated From Caatinga Soil Maria Carolina de Albuquerque Wanderley 1,2 , Jose Manoel Wanderley Duarte Neto 2 , Carolina de Albuquerque Lima 3 , Sara Isabel da Cruz Silverio 4 , Jose Luiz de Lima Filho 2 , Jose Antonio Couto Teixeira 4 , Ana Lucia Figueiredo Porto 5* 1 Department of Molecular Sciences, Federal Rural University of Pernambuco (UFRPE), Av. Dom Manoel de Medeiros, s/n, 52171-900, Recife, PE, Brazil. 2 Laboratory of Immunopathology Keizo Asami (LIKA), Federal University of Pernambuco, Av. Prof. Moraes Lins do Rego, s/n, 50670-901, Recife, PE, Brazil. 3 Faculty of Science, Education and Technology of Garanhuns, University of Pernambuco-UPE, Av. Capitão Pedro Rodrigues, n° 105, Garanhuns, PE, Brazil. 4 Centre of Biological Engineering, University of Minho, Campus Gualtar, 4710-057, Braga, Portugal. 5 Department of Morphology and Animal Physiology, Federal Rural University of Pernambuco, Av. Dom Manoel de Medeiros, s/n, 52171-900, Recife, PE, Brazil. ARTICLE INFO ABSTRACT Article history: Received on: 14/06/2016 Revised on: 14/07/2016 Accepted on: 30/07/2016 Available online: 26/08/2016 A new Penicillium sp. strain isolated from the soil of Caatinga, a Brazilian Biome (UCP 1286) was selected for collagenase production. Fermentation system allowing obtention of collagenolytic activity about 2.7 times higher than existing data, with the highest values of collagenolytic and specific activity (379.80 U/mL, 1460.77 U/mg, respectively), after 126 hours. Applying a factorial design, enzyme production was increased by about 65% compared to the preliminary results. The factorial design demonstrated the existence of two factors with statistical significance on the production of the enzyme: pH and temperature, both with negative effects. Enzyme was found to be more active at pH 9.0 and 37 °C, and also to be very stable in comparison with the collagenase produced by other microorganisms. The enzyme seems to belong to collagenolytic serine proteases family. Concerning the substrate specificity, it was observed that the highest enzyme activity corresponds to azocoll, there was no relevant activity on azocasein and the enzyme showed to be more specific to type V collagen and gelatin than the commercial colagenase produced by Clostridium histolyticum. Major band observed at electrophoresis was approximately 37 kDa. Zymogram analysis confirmed the collagenolytic activity. All data indicates this enzyme as promising biotechnology product. Key words: Collagenolytic; Enzymes; Factorial Design; Fermentation; Filamentous Fungi; Specificity. 1. INTRODUCTION Collagen is the major fibrous element of skin, bones, tendons, cartilage, blood vessels and teeth found in all animals [1,2]. Collagen is found in connective tissues, making up approximately 30% of the protein in human body [3,4]. Because of the rigid structure of collagen (three helically wound polypeptide fibrils) its degradation is restricted to a few proteases [2]. Collagenases are specific enzymes that can hydrolyze both * Corresponding Author Department of Morphology and Animal Physiology, Federal Rural University of Pernambuco, Av. Dom Manoel de Medeiros, s/n, 52171- 900, Recife, PE, Brazil. Email: analuporto@yahoo.com.br, Telephone number: +55 81 33206391, Fax number: +51 81 21268485 native and denatured collagens [5,6]. These enzymes can degrade native triple helix of collagen to small fragments and play an important role in connective tissue metabolism [7,8]. Collagenolytic enzymes are a kind of proteases that are related to various physiological and pathological processes and have several applications in industry, medicine and biotechnology [6,9–11]. With biotechnology accelerated growth, applications of proteases have expanded to new areas such as clinical, medicinal and analytical chemistry [12]. Among various sources of proteases, those produced by microorganisms play an important role in biotechnological processes and are used with increasing frequency, as large amounts of these enzymes can be produced quickly and at low cost [13]. Search for microbial collagenases has been increasing due to their wide application, as they are able to cleave collagen helix at multiple sites, while mammalian collagenases cleave at a single site [1,14].