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

• Published in: Applied sciences 2021-01, Vol.11 (4), p.1456
• Main Authors: Hayakawa, Yusuke, Nakayama, Ryoichi, Namiki, Norikazu, Imai, Masanao
• Format: Article
• Language: English
• Subjects: Accurel MP100; Aqueous solutions; Enzymatic activity; Enzyme activity; Enzymes; Ethanol; Fatty acids; Glutaraldehyde; Hydrocarbons; Hydrolysis; Hydrophobicity; Immobilization; Industrial applications; Investigations; Microemulsions; Moisture content; Molecular structure; Phospholipase; Phospholipase A1; Phospholipids; Reactivity; Scanning electron microscopy; Solvents; Water content; Yield
• ISSN: 2076-3417; 2076-3417
• DOI: 10.3390/app11041456

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.