This study aims at optimizing medium-chain-length Polyhydroxyalkanoate (MCL-PHA) production from Pseudomonas aeruginosa 13003 culture using the statistical design techniques of 2-level factorial design and RSM. Two-level factorial design with eight variables viz. oleic acid, (NH4)(2)PO4, incubation time, trace metal solution, MgCl2, K2HPO4, KH2PO4 and inoculum was performed to screen for nutrients that were significantly affecting MCL-PHA production. After the values for the less-influential variables were fixed, oleic acid, (NH4)(2)PO4, K2HPO4 and KH2PO4 having high t-values were selected to study their optimal relationship for maximal PHA production using 2(4) Central Composite Design. The optimal composition of the medium was found to be at 7.5 g L-1 oleic acid, 0.4 g L-1 (NH4)(2)PO4, 3.0 g L (1) K2HPO4 and 1.5 g L (1) KH2PO4 which gave 60% of CDW as MCL-PHA. This optimization resulted in a 1.6 times increase in the PHA content (60% mass fraction in contrast to 36%). On analysis carbon to nitrogen ratio of 19 was found to be optimum for PHA production. This is the first report on the use of CCD to improve MCL-PHA production from P. aeruginosa. (C) 2013 Elsevier Ltd. All rights reserved.