We have investigated biosynthetic characteristics of two newly isolated Pseudomonas sp. 1 CNM-PsB-01 and Pseu domonas sp. 2 CNM-PsB-02 strains, in parallel with biotechnological tools oriented for obtaining enzymatic fractions with an elevated lipolytic activity. Based on rational design and mathematical optimization we suggested four new nutritive media, containing secondary organic resources, finally leading to prominent levels of lipo-enzymatic complexes. Production costs of these lipolytic complexes are reduced due to utilization of the aqueous supernatant resulting after collection of the primary Dunaliella salina biomass. We concluded that via combination of modern principles of biotechnology, and making use of secondary nutrient resources, like sterilized aqua fraction of D. salina, it is possible to obtain two major products in one biotech cycle, that is Dunaliella salina biomass and the lipolytic complex resulting from cultivation of Pseudomonas on the residual media.