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

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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.

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Funding

We are thankful for funding from the National Natural Science Foundation of China for Young Scholars, grant/award number: 22108227; Shaanxi Provincial Natural Science Foundation, grant/award number: 2023-JC-YB-092; the Shaanxi Association for Science and Technology for Young Scholars, grant number: 20220201; and the Scientific research projects of Shaanxi Education Department, grant/award number: 2021JQ-750.

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Authors and Affiliations

  1. Shaanxi Key Laboratory of Catalysis, School of Chemistry and Environment Science, Shaanxi University of Technology, No.1 Dongyihuan Road, Hanzhong, Shaanxi, 723001, China

    Xia Zhao & Bo Liu

  2. College of Food Science and Engineering, Northwest University, Xi’an, China

    Meijing Guo, Xian Li, Binglin Li & Jiao Wang

  3. Biochemistry Center (BZH), Heidelberg University, 69120, Heidelberg, Germany

    Jiao Wang

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  1. Xia Zhao
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  2. Meijing Guo
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  3. Xian Li
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  4. Bo Liu
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  5. Binglin Li
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Correspondence to Binglin Li or Jiao Wang.

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Zhao, X., Guo, M., Li, X. et al. Immobilization of Bio-imprinted Phospholipase D and Its Catalytic Behavior for Transphosphatidylation in the Biphasic System. Appl Biochem Biotechnol 195, 7808–7820 (2023). https://doi.org/10.1007/s12010-023-04528-4

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  • DOI: https://doi.org/10.1007/s12010-023-04528-4

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