Engineering of a Microbial Cell Factory for the Extracellular Production of Catalytically Active Phospholipase A of .

Phospholipase A (PLA) from is a lipolytic enzyme used in a wide range of industrial applications including production of lysolecithins and enzymatic degumming of edible oils. We have therefore investigated expression and secretion of PLA in two workhorse microbes, and . The PLA was produced to an activity of 0.517 ± 0.012 U/ml in the culture broth of the recombinant . On the other hand, recombinant BL21 star (DE3), overexpressing the authentic PLA (P-PLA2), showed activity of 17.0 ± 1.3 U/ml in the intracellular fraction and 21.7 ± 0.7 U/ml in the culture broth. The extracellular PLA activity obtained with the recombinant system was 3.2-fold higher than the corresponding value reached in a previous study, which employed recombinant BL21 (DE3) overexpressing codon-optimized PLA2. Finally, we observed that the extracellular PLA from the recombinant P-PLA culture was able to hydrolyze 31.1 g/l of crude soybean lecithin, an industrial substrate, to a conversion yield of approximately 95%. The newly developed -based PLA expression system led to extracellular production of PLA to a productivity of 678 U/l·h, corresponding to 157-fold higher than that obtained with the -based system. This study will contribute to the extracellular production of a catalytically active PLA.