Studies on the Zymogram Method for the Detection of Pectinolytic Activities Using CTAB Noomen Hadj-Taieb & Hajer Tounsi & Amina Chabchoub & Najla Abid & Ali Gargouri Received: 25 January 2011 /Accepted: 5 September 2011 / Published online: 21 September 2011 # Springer Science+Business Media, LLC 2011 Abstract Zymogram analysis is a useful tool for the identification of several enzymes. The present study was undertaken to investigate the efficiency gains from the characterization of pectic enzymes on zymograms by staining of pectin–agarose overlays using cetyl trimethyl ammonium bromide also known as cetrimide or CTAB. The method is based on the fact that the enzymatic hydrolysis of the pectic substrates included in the agarose matrix gel inhibited their precipitation by CTAB, leading to the appearance of cleared zones in front of the pectin hydrolases and lyases. Conversely, esterases led to the increase of pectin precipitation. Fungal pectinolytic enzymes were separated on sodium dodecyl sulfate-polyacrylamide gel electro- phoresis and subjected to the zymogram detection technique, using two pectin substances, namely citrus pectin and polygalacturonic acid. Overall, the findings presented in the current study indicate that several elements (ions, salts, pH, temperature, chelators, and reducing agents) may significantly affect the results of zymogram analysis and can, therefore, be employed to enhance the discriminatory and operational potential of the analysis in terms of accurate discrimination between several pectinolytic activities involved and effective implementation of the purification procedures required in the process. Keywords Zymogram . Pectinase activities . Cetyl trimethyl ammonium bromide (CTAB) . CT1 mutant: Penicillium occitanis Introduction The hydrolysis of pectin backbones can be achieved through the synergistic action of several enzymes, including depolymerases (polymethylgalacturonases, polygalacturonases, pectate lyases, and pectin lyases) and pectin methylesterases [1]. Several organisms are able to produce pectin-degrading enzymes, which are, in fact, widely used in a variety of food Appl Biochem Biotechnol (2011) 165:1652–1660 DOI 10.1007/s12010-011-9384-y N. Hadj-Taieb (*) : H. Tounsi : A. Chabchoub : N. Abid : A. Gargouri Laboratoire de Valorisation de la Biomasse et Production des Protéines chez les Eucaryotes, Centre de Biotechnologie de Sfax, Université de Sfax, Route Sidi Mansour km 6, BP 1177, 3038 Sfax, Tunisia e-mail: noomen.hadjtaib@cbs.rnrt.tn