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Recovery of Extracellular Lipolytic Enzymes from Macrophomina phaseolina by Foam Fractionation with Air

DOI: 10.1155/2013/897420

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Macrophomina phaseolina was cultivated in complex and simple media for the production of extracellular lipolytic enzymes. Culture supernatants were batch foam fractionated for the recovery of these enzymes, and column design and operation included the use of P 2 frit (porosity 40 to 100?μm), air as sparging gas at variable flow rates, and Triton X-100 added at the beginning or gradually in aliquots. Samples taken at intervals showed the progress of the kinetic and the efficiency parameters. Best results were obtained with the simple medium supernatant by combining the stepwise addition of small amounts of the surfactant with the variation of the air flow rates along the separation. Inert proteins were foamed out first, and the subsequent foamate was enriched in the enzymes, showing estimated activity recovery (R), enrichment ratio (E), and purification factor (P) of 45%, 34.7, and 2.9, respectively. Lipases were present in the enriched foamate. 1. Introduction Macrophomina phaseolina (Tassi) Goid. [1], the only species of its gender, is a phytopathogenic filamentous fungus, belonging to the anamorphic Ascomycota, Botryosphaeriaceae family [2]. It was recently described as possessing “tools to kill” [3] due to its genome providing a diversified arsenal of enzymatic and toxin tools to destroy the host plants, a capacity that is confirmed by its ability to infect over 500 different plant species [4]. Thus, to presume that Macrophomina phaseolina is able to produce several enzymes suitable for industrial applications is a reasonable hypothesis. To this purpose, many studies on its cell wall degrading hydrolases were performed [5–12]. However, M. phaseolina produces several other extracellular enzymes [13] of potential industrial use, among them lipolytic enzymes, which are excreted into the culture media in different amounts depending on the strain and incubation conditions. No studies were found attempting to purify these lipolytic enzymes. Several processes in the food industry, as well as environmental and industrial biotechnological applications, use enzymes as biocatalysts [14] due to their many advantages over chemical catalysts: the ability to function under relatively mild conditions of temperature, pH, and pressure; their specificity and, in some cases, their stereoselectivity. In addition, they do not produce unwanted byproducts [15]. Lipases are of particular interest because of their many applications in oleochemistry, organic synthesis, the detergent industry, and nutrition [16], and there is constant search for new options [17]. Eco-friendly

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