Promising Immobilization of Industrial-Class Phospholipase A1 to Attain High-Yield Phospholipids Hydrolysis and Repeated Use with Optimal Water Content in Water-in-Oil Microemulsion Phase

In this study, we maximized the reactivity of phospholipids hydrolysis with immobilized industrial-class phospholipase A(1) (PLA(1)) at the desired water content in the water-in-oil (W/O) microemulsion phase. The optimal hydrophobic-hydrophilic condition of the reaction media in a hydrophobic enzyme reaction is critical to realize the maximum yields of enzyme activity of phospholipase A(1). It was attributed to enzymes disliking hydrophobic surroundings as a special molecular structure for reactivity. Immobilization of PLA(1) was successfully achieved with the aid of a hydrophobic carrier (Accurel MP100) combination with the treatment using glutaraldehyde. The immobilized yield was over 90% based on simple adsorption. The hydrolysis reaction was kinetically investigated through the effect of glutaraldehyde treatment of carrier and water content in the W/O microemulsion phase. The initial reaction rate increased linearly with an increasing glutaraldehyde concentration and then leveled off over a 6% glutaraldehyde concentration. The initial reaction rate, which was predominantly driven by the water content in the organic phase, changed according to a typical bell-shaped curve with respect to the molar ratio of water to phospholipid. It behaved in a similar way with different glutaraldehyde concentrations. After 10 cycles of repeated use, the reactivity was well sustained at 40% of the initial reaction rate and the creation of the final product. Accumulated yield after 10 times repetition was sufficient for industrial applications. Immobilized PLA(1) has demonstrated potential as a biocatalyst for the production of phospholipid biochemicals.

Automated summary

This study focused on maximizing the reactivity of phospholipids hydrolysis by immobilizing industrial-class phospholipase A(1) in the water-in-oil microemulsion phase; the hydrophobic-hydrophilic condition of the reaction media was found to be critical for achieving maximum enzyme activity yields. Immobilized PLA(1) demonstrated sustained reactivity after multiple uses, showing potential for industrial applications in producing phospholipid biochemicals.