Comparison of Lipase Production by Enterococcus faecium MTCC 5695 and Pediococcus acidilactici MTCC 11361 Using Fish Waste as Substrate: Optimization of Culture Conditions by Response Surface Methodology
A medium using fish waste as substrate was designed for production of lipase by Enterococcus faecium MTCC 5695 and Pediococcus acidilactici MTCC 11361. Medium components and culture conditions (fish waste protein hydrolysate (FWPH) concentration, fish waste oil (FWO) concentration, pH, temperature, and fermentation time) which affect lipase production were screened using factorial (5 factors ？ 2 levels) design of which FWPH concentration, FWO concentration, and fermentation time showed significance ( ). The levels of these factors were optimized further by Box-Behnken design using response surface methodology (RSM). Optimized conditions were found to be 5%？v/v FWO, 0.15？mg/mL FWPH and 24？h of fermentation time for MTCC 5695, and 4%？v/v FWO, 0.15？mg/mL FWPH and 24？h of fermentation for MTCC 11361, which were further validated. Under optimized conditions, MTCC 5695 and MTCC 11361 showed 3.15- (543.63 to 1715？U/mL) and 2.3- (214.74 to 493？U/mL) fold increase in lipase production, respectively, as compared to unoptimized conditions. 1. Introduction Lipases (triacylglycerol acylhydrolases EC 3.1.1.3) are a class of serine hydrolases which catalyze the hydrolysis of triglycerides to glycerol and free fatty acids over oil-water interface [1]. In addition, lipases catalyze the hydrolysis and transesterification of other esters as well as the synthesis of esters and exhibit enantioselective properties [2]. These unique properties of lipases make them a very important enzyme of industrial significance. Lipases are used in chemical processing, dairy industries for improvement of flavour, paper industries, oleochemical industries, pharmaceuticals, synthesis of surfactants, detergent industries, leather industries, and polymer synthesis [3, 4]. Lipases are produced by plants, animals, and microbes but only microbial lipases are found to be industrially important since they are diversified in their enzymatic properties and substrate specificity [5]. Bacterial lipases that are of commercial importance are obtained from Achromobacter, Alcaligenes, Arthrobacter, Bacillus, Burkholderia, Chromobacterium, and Pseudomonas [6, 7]. Lactic Acid Bacteria (LAB) are generally considered to be weakly lipolytic, as compared to other groups of microorganisms. The lipolytic activity by LAB plays an important role in the determination of special aroma of different cheeses [8, 9]. Since they are considered as generally recognised as safe (GRAS), they are used extensively as starter cultures in food and feed industries [10]. Although there are reports on lactic acid bacterial lipase
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