Microbial enzymes from extremophilic regions such as hot spring serve as an important source of various stable and valuable industrial enzymes. The present paper encompasses the modeling and optimization approach for production of halophilic, solvent, tolerant, and alkaline lipase from Staphylococcus arlettae through response surface methodology integrated nature inspired genetic algorithm. Response surface model based on central composite design has been developed by considering the individual and interaction effects of fermentation conditions on lipase production through submerged fermentation. The validated input space of response surface model (with value of 96.6%) has been utilized for optimization through genetic algorithm. An optimum lipase yield of 6.5?U/mL has been obtained using binary coded genetic algorithm predicted conditions of 9.39% inoculum with the oil concentration of 10.285% in 2.99?hrs using pH of 7.32 at 38.8°C. This outcome could contribute to introducing this extremophilic lipase (halophilic, solvent, and tolerant) to industrial biotechnology sector and will be a probable choice for different food, detergent, chemical, and pharmaceutical industries. The present work also demonstrated the feasibility of statistical design tools integration with computational tools for optimization of fermentation conditions for maximum lipase production. 1. Introduction Hydrolases particularly lipases (triacylglycerol acylhydrolases, EC 3.1.1.3) from extremophilic microorganisms are experiencing a growing demand, due to their versatile catalytic activities (regio- and enantioselectivity) coupled multifold industrial applications [1]. Among different sources, microbial lipases have gained special industrial attention due to their stability, selectivity, broad substrate specificity, and their cost-effective production. The extracellular bacterial lipases are of considerable commercial importance, due to their substrate specificity, their ability to function in extreme environments, and their bulk production being much easier. Currently bacterial lipases are of great demand because they tend to have neutral or alkaline pH optima and are often thermostable [2, 3]. Lipases from extremophiles are capable of functioning in presence of salts, oxidizing agents, and organic solvents and can withstand the harsh industrial conditions which may permit their use in some specialized industrial applications, such as novel substrates catalysis reactions [4]. Production of lipases through submerged fermentation (SmF) avoids the unwanted metabolites production
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