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

• Main Authors: Yusuke Hayakawa, Ryoichi Nakayama, Norikazu Namiki, Masanao Imai
• Format: Article
• Language: English
• Published: MDPI AG 2021-02-01
• Series: Applied Sciences
• Subjects: immobilization; phospholipase A₁; hydrolysis; Accurel MP100; water content
• Online Access: https://www.mdpi.com/2076-3417/11/4/1456

In this study, we maximized the reactivity of phospholipids hydrolysis with immobilized industrial-class phospholipase A₁ (PLA₁) 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 A₁. It was attributed to enzymes disliking hydrophobic surroundings as a special molecular structure for reactivity. Immobilization of PLA₁ 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 PLA₁ has demonstrated potential as a biocatalyst for the production of phospholipid biochemicals.