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. 2023 Dec;195(12):7808-7820.
doi: 10.1007/s12010-023-04528-4. Epub 2023 Apr 24.

Immobilization of Bio-imprinted Phospholipase D and Its Catalytic Behavior for Transphosphatidylation in the Biphasic System

Xia Zhao  1 , Meijing Guo  2 , Xian Li  2 , Bo Liu  1 , Binglin Li  3 , Jiao Wang  4   5
Affiliations

Immobilization of Bio-imprinted Phospholipase D and Its Catalytic Behavior for Transphosphatidylation in the Biphasic System

Xia Zhao et al. Appl Biochem Biotechnol. 2023 Dec.

Abstract

Phospholipase D (PLD) with the higher transphosphatidylation activity was screened from Streptomyces sp. LD0501 basing on the protoplast mutagenesis technology. Then, it was successfully bio-imprinted to form a hyperactivated structure and rigidified by the intramolecular cross-linking, which was immobilized on the nonporous nanoscale silica. Characterization techniques were employed to investigate the structure and physicochemical properties of the catalysts, including Fourier transform infrared (FTIR) spectra and scanning electron microscopy (SEM) analysis. Transphosphatidylation activity and selectivity were improved significantly when immobilized PLD was used. The maximum yield for the production of phosphatidylserine (PS) reached 97% and the side reaction, the hydrolysis, was minimized. These results were further confirmed by the nuclear magnetic resonance (NMR) and mass spectrometry (MS) analysis. The imprint-induced characteristics of PLD was successfully "remembered" even in the present of much water. In addition, this immobilized hyperactivated PLD showed the excellent operational stabilities and environmental tolerances.

Keywords: Bio-imprint; Immobilization; Mutagenesis; Phosphatidylserine; Phospholipase D.

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