Cell wall-associated lipases are attractive biocatalysts as they reduce the downstream production cost and enable easy recovery and reuse as compared to secretory (free) enzymes. Here, we investigated the effect of the oxygen carrier perfluorodecalin (PFC) on the production of lipase fractions from Yarrowia lipolytica, with emphasis on a cell wall-associated biocatalyst obtained by cell ultrasound disruption (hereafter LipImDebris). Two Yarrowia lipolytica strains were considered for that purpose: the wild-type IMUFRJ 50682 and genetically modified JMY1108 strain producing only the Lip8 isoform (JMY1108, Lip2D, Lip7D). Different crude vegetable oils and waste frying oils (WFO) were evaluated as carbon source and inducer for lipase synthesis. LipImDebris activity was not detected in JMY1108 strain, suggesting that it predominantly corresponds to the Lip2 isoform in the wild-type strain. For crude vegetable oils, the highest LipImDebris activity was observed after 15 h, with cottonseed oil reaching 718 U g⁻¹. With WFO, LipImDebris activity was reduced relative to the corresponding crude vegetable oils, yet measurable activity remained. The addition of 20% (v/v) PFC in the culture medium markedly enhanced LipImDebris formation, increasing activity by 9-fold with olive oil, 17-fold with cottonseed oil, and 2-fold with soybean WFO. LipImDebris produced from Y. lipolytica culture with soybean WFO in the presence of PFC retained substantial hydrolytic activity for up to four reuse cycles. Overall, coupling WFO with improved oxygen availability is a promising strategy for producing reusable, naturally immobilized lipases with industrial applications.