In this work, an induced pre-saturation tower (IPST) for oil–water separation was built on a semi-industrial scale, based on experimental results obtained on a laboratory scale prototype. The main strategy for generating these criteria was to increase the efficiency of the bench scale prototype, which is limited by conditions of low levels of automation and control, with the use of a biosurfactant as an auxiliary collector. The validation of the developed criteria allowed the construction of an IPST with three stages, all fed with previously saturated effluents. The IPST was built in stainless steel, with multistage centrifugal pumps and adapted to generate microbubbles without the use of saturation tanks or compressors. The most relevant operational parameters were selected using a fractional factorial design, while a central composite rotatable design (CCRD) followed by the application of the desirability function allowed to optimize the conditions for partial and global variables, the latter with desirability of 95%. A nominal flow rate of approximately 1000 L·h−1, a recycle flow rate of 450 L·h−1, a scraper rotation speed of 80 rpm, an average pressure of the microbubble pumps of 11 bar, and an effluent temperature from IPST of about 38 °C ensured optimized operation for the proposed technological development.