The Use of Response Surface Methodology as a Statistical Tool for Media Optimization in Lipase Production from the Dairy Effluent Isolate Fusarium solani
The optimization of extracellular lipase production by Fusarium isolani strain SKWF7 isolated from dairy wastewater was carried out in this study. Initially, the physicochemical factors significantly influencing enzyme production were studied by varying one-factor-at-a-time (OFAT). A mesophilic temperature of 40°C, alkaline pH of 8, and incubation period of 72 hours were found to be the optimal conditions for lipase production. Among the media components, the disaccharide sucrose acted as the best carbon source; palm oil as the best inducing lipid substrate; casein and (NH4)2SO4 as the best organic and inorganic nitrogen sources; Ca2+ ion as the best trace element. In the next phase of work, statistical optimization of medium components was performed by employing the Box-Behnken design of Response Surface Methodology (RSM). The optimum concentrations of three significant factors, namely, palm oil, (NH4)2SO4, and CaCO3 were determined by this method to be 5% (v/v), 5.5？g/L, and 0.1？g/L, respectively. RSM-guided design of experiments resulted in a maximum lipase production of 73.3？U/ml, which is a 1.7-fold increase in comparison with that obtained in the unoptimized medium. These results point towards the success of the model in developing a process for the production of lipase, an enzyme of enormous industrial significance. 1. Introduction Lipases are enzymes that belong to the class of hydrolases and are involved in catalyzing the hydrolysis of triglycerides to fatty acids and glycerol, this reaction occurs at the oil-water interface [1]. Besides, they are also capable of catalyzing the reverse reaction, that is, ester synthesis, in water-restricted environments [2]. Transesterification and resolution of racemic mixtures are also reactions that could be facilitated by lipases. Such a versatile nature has paved the way for their application in diversified industries including food, dairy, cosmetic, detergent, and pharmaceuticals [3]. The environmental applications of the enzyme are also innumerable, where they break down fat, oil, and greasy material in the wastewater [4]. These constituents could pose problems in the sewers as well as the treatment plant, impeding oxygen transfer in aerobic biological treatment systems. These are some of the underlying reasons for a sustained interest in the enzyme. Lipases can be procured from plant, animal [5–8], and microbial sources. However, microbial lipases have gained increasing attention due to their stability and high substrate specificity [9, 10]. In the microbial community, lipolytic activity is exhibited by
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