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Volume 14
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10.3390/biom14040430
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Article

Whole-Cell Display of Phospholipase D in Escherichia coli for High-Efficiency Extracellular Phosphatidylserine Production

by
Baotong Sun
,
Zhongchen Li
,
Yanhong Peng
,
Fei Wang
,
Yibin Cheng
,
Yang Liu
* and
Lixin Ma
*
State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei University, Wuhan 430062, China
*
Authors to whom correspondence should be addressed.
Biomolecules 2024, 14(4), 430; https://doi.org/10.3390/biom14040430
Submission received: 4 February 2024 / Revised: 25 March 2024 / Accepted: 28 March 2024 / Published: 2 April 2024
(This article belongs to the Section Molecular Medicine)
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Abstract

Phospholipids are widely utilized in various industries, including food, medicine, and cosmetics, due to their unique chemical properties and healthcare benefits. Phospholipase D (PLD) plays a crucial role in the biotransformation of phospholipids. Here, we have constructed a super-folder green fluorescent protein (sfGFP)-based phospholipase D (PLD) expression and surface-display system in Escherichia coli, enabling the surface display of sfGFP-PLDr34 on the bacteria. The displayed sfGFP-PLDr34 showed maximum enzymatic activity at pH 5.0 and 45 °C. The optimum Ca2+ concentrations for the transphosphatidylation activity and hydrolysis activity are 100 mM and 10 mM, respectively. The use of displayed sfGFP-PLDr34 for the conversion of phosphatidylcholine (PC) and L-serine to phosphatidylserine (PS) showed that nearly all the PC was converted into PS at the optimum conditions. The displayed enzyme can be reused for up to three rounds while still producing detectable levels of PS. Thus, Escherichia coli/sfGFP-PLD shows potential for the feasible industrial-scale production of PS. Moreover, this system is particularly valuable for quickly screening higher-activity PLDs. The fluorescence of sfGFP can indicate the expression level of the fused PLD and changes that occur during reuse.
Keywords: phosphatidylserine production; visible sfGFP; phospholipase D; cell-surface display; reused phosphatidylserine production; visible sfGFP; phospholipase D; cell-surface display; reused

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MDPI and ACS Style

Sun, B.; Li, Z.; Peng, Y.; Wang, F.; Cheng, Y.; Liu, Y.; Ma, L. Whole-Cell Display of Phospholipase D in Escherichia coli for High-Efficiency Extracellular Phosphatidylserine Production. Biomolecules 2024, 14, 430. https://doi.org/10.3390/biom14040430

AMA Style

Sun B, Li Z, Peng Y, Wang F, Cheng Y, Liu Y, Ma L. Whole-Cell Display of Phospholipase D in Escherichia coli for High-Efficiency Extracellular Phosphatidylserine Production. Biomolecules. 2024; 14(4):430. https://doi.org/10.3390/biom14040430

Chicago/Turabian Style

Sun, Baotong, Zhongchen Li, Yanhong Peng, Fei Wang, Yibin Cheng, Yang Liu, and Lixin Ma. 2024. "Whole-Cell Display of Phospholipase D in Escherichia coli for High-Efficiency Extracellular Phosphatidylserine Production" Biomolecules 14, no. 4: 430. https://doi.org/10.3390/biom14040430

APA Style

Sun, B., Li, Z., Peng, Y., Wang, F., Cheng, Y., Liu, Y., & Ma, L. (2024). Whole-Cell Display of Phospholipase D in Escherichia coli for High-Efficiency Extracellular Phosphatidylserine Production. Biomolecules, 14(4), 430. https://doi.org/10.3390/biom14040430

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