Lipases are biocatalysts with a significant potential to enable a shift from current pollutant manufacturing processes to environmentally sustainable approaches. The main reason of this prospect is their catalytic versatility as they carry out several industrially relevant reactions as hydrolysis of fats in water/lipid interface and synthesis reactions in solvent-free or non-aqueous media such as transesterification, interesterification and esterification. Because of the outstanding traits of Rhizopus oryzae lipase (ROL), 1,3-specificity, high enantioselectivity and stability in organic media, its application in energy, food and pharmaceutical industrial sector has been widely studied. Significant advances have been made in the biochemical characterisation of ROL particularly in how its activity and stability are affected by the presence of its prosequence. In addition, native and heterologous production of ROL, the latter in cell factories like Escherichia coli, Saccharomyces cerevisiae and Komagataella phaffii (Pichia pastoris), have been thoroughly described. Therefore, in this review, we summarise the current knowledge about R. oryzae lipase (i) biochemical characteristics, (ii) production strategies and (iii) potential industrial applications.

Grup de Recerca Enginyeria de Bioprocesos i Biocatàlisi AplicadaA series of lipase-catalysed transesterification experiments were carried out to study the effect of the presence of free fatty acids on synthesis reaction rate and the stability of the biocatalyst, and also to elucidate the underlying mechanism, which remains a subject of debate. Based on the results, the reaction rate and biocatalyst stability increased with increasing content in free fatty acids of the reaction mixture. Also, tests carried out with a mixture of triolein and linoleic acid revealed that the transesterification mechanism is a combination of direct alcoholysis of triacylglycerols and a twostep reaction involving hydrolysis of acylglycerols and further esterification of previously released free fatty acids. The time course of triacylglycerols and diacylglycerols revealed that the enzyme is similarly selective for both types of substrate