Intracellular lipase producer screened from the library available in the laboratory, identified through 16S rRNA as Pseudomonas stutzeri, was studied for maximum enzyme production in shake flask. The work was intended to evaluate the effect of different physicochemical factors like carbon, nitrogen, metal ions, surfactant, inoculum, pH, temperature, agitation, and aeration on lipase production. Optimized media showed 1.62-fold increase in lipase production when compared to basal media. Scale-up of lipase in in situ bioreactor showed reduction in fermentation time in both basal and optimized media, giving 41 and 99？U/mg of lipase activity after 48？h of fermentation. 1. Introduction Lipases are carboxylesterases (E.C. 3.1.1.3) that catalyze both hydrolysis and synthesis of esters formed from glycerol and fatty acids [1, 2]. They are the serine hydrolases which generally act at the oil-water interface. Structural attributes include an α/β hydrolase fold and the active site of the enzyme is generally composed of catalytic triad which is surrounded by a consensus sequence of Gly-x-Ser-x-Gly [3]. Lipases are currently attracting an enormous attention because of their wide range of applications in natural sciences and biotechnology [4]. They often retain their biological activity in organic solvents and display exquisite chemo-, regio-, and enantioselectivity that provide them an edge over other classes of biocatalysts. Due to their properties they have become almost indispensable biocatalysts in various industrial sectors such as the agrochemical, pharmaceutical, detergent, and food industries [5]. The number of lipases has increased since the 1980s and they are studied from microorganisms such as Pseudomonas, Bacillus, Achromobacter, Alcaligenes, Arthrobacter, and Chromobacterium [6–8]. It is a well known fact that variation in media components has a significant role in influencing microbial metabolic activities and thereby enzyme production. Generally, high enzyme titre has been achieved by culture medium changes in terms of various possible physicochemical components that are likely to influence the enzyme production and growth of microorganisms in time course fermentation [9]. Optimization and evaluation of each ingredient that constitutes a cultivation medium is the foundation in classical approach. Therefore optimizing best possible conditions for maximum production of enzyme is still an important and imperative area in enzymology. In the present study, lipase producer Pseudomonas stutzeri MTCC 5618 was studied for maximum enzyme production through
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