Lipases are among the most significant biocatalysts that constitute the third most important group of enzymes due to their vast range of applications. The present research represents the first attempt to optimize the growth medium constituents to increase the production of recombinant lipase in Pseudomonas aeruginosa SDK-6. One factor at a time (OFAT) revealed castor oil, yeast extract, and ammonium nitrate as the most significant medium components affecting the recombinant lipase production. Further optimization via response surface methodology (RSM) resulted in lipase production of 115.50 U/mL with 0.5% (v/v) castor oil, 0.2% (w/v) yeast extract, and 0.1% (w/v) ammonium nitrate at pH 7. Statistical validation of the observed value via ANOVA revealed an F value of 117.02 at p <
0.01, with an R