Integrated enzymatic processes for next-generation biodiesel production

Enzymatic biodiesel production enables process-integrated transformation of low-quality, heterogeneous feedstocks through synergistic biocatalysis. From that perspective, this review consolidates advances involving lipases, phospholipases, and multi-enzyme systems applied to transesterification, esterification, and enzymatic degumming (EDG). Commercial lipases such as Novozym® 435, Lipozyme® TL-IM/RM-IM, and Eversa® Transform 2.0 (ET 2.0) achieve biodiesel yields above 90%, with immobilized preparations improving stability and enabling reuse for 15–20 cycles. Magnetic nanoparticle immobilization of ET 2.0, for example, increases specific activity twofold and maintains 96.7% conversion over six cycles, while Rhizopus oryzae lipase immobilized on octadecyl methacrylate achieves a 234% activity increase compared with the free enzyme. Phospholipases play a central role in mitigating inhibition in crude oils. EDG with Quara® LowP or Purifine® 3 G reduces phosphorus from 900 to 10 ppm, achieving > 90% hydrolysis and increasing diacylglycerol availability, thereby facilitating downstream lipase catalysis. Lipase-phospholipase synergy enables one-pot processes, where simultaneous degumming and transesterification achieve biodiesel yields of 86–97.5%. Combi-lipases further enhance conversion of multifunctional substrates, reaching yields of 75–99% within 2–7 h. Innovations such as enzyme co-immobilization on mesoporous SBA-15, magnetic biochar, and CaCO₃ supports increase selectivity and stability. Furthermore, hybrid reactor designs and downstream integration optimize separation and purification efficiency. Despite economic challenges, process intensification through multi-enzyme systems and advanced immobilization offers a clear pathway toward operational efficiency, enabling high catalytic retention over multiple cycles. Collectively, these advances support the development of scalable and sustainable enzymatic platforms for next-generation biodiesel production.