Characterization and heterologous expression of plasmalogen synthase MeHAD from Megasphaera elsdenii.

Furong Zhang, Zhaonan Yang, Yulong Zhou, Binqiang Wang, Zhenming Xie, Ning Yu, Jie Zhao, Howard Goldfine, Shang Dai, Genlin Zhang, Bing Tian

Biochimica et biophysica acta. Molecular and cell biology of lipids 2023 Sep

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Plasmalogens (Pls) are vinyl-ether bond-containing glycerophospholipids or glycosyl diradyl glycerols, and are of great importance in the physiological functions and stability of cell membrane. Here, we identified and characterized that the plasmalogen synthase MeHAD from anaerobic Megasphaera elsdenii was responsible for vinyl-ether bond formation. Different from the 2-hydroxyacyl-CoA dehydratase (HAD) family plasmalogen synthase PlsA-PlsR which are encoded by two genes in Clostridium perfringens, the HAD homolog (MeHAD) encoded by a single gene MELS_0169 was found in M. elsdenii. By heterologous expression of the MeHAD gene into a nonplasmalogen-producing Escherichia coli strain, the expressed MeHAD was found to be located in the cell membrane region. Plasmalogens were detected in the recombinant strain using GC-MS and LC-MS, demonstrating that MeHAD was the key enzyme for plasmalogen synthesis. Moreover, the synthesized plasmalogens could enhance the oxidative stress-resistance and osmotic pressure-resistance of the recombinant strain, probably due to the ROS scavenging and decreased membrane permeability by the plasmalogens, respectively. The four-cysteine (Cys125, Cys164, Cys445 and Cys484) site-mutant of MeHAD, which were predicted binding to the [4Fe-4S] cluster, was unable to synthesize plasmalogens, indicating that the cysteines are important for the catalytic activity of MeHAD. Our results revealed the single gene encoded plasmalogen synthase in M. elsdenii and established a recombinant E. coli strain with plasmalogen production potential. Copyright © 2023 Elsevier B.V. All rights reserved.

Citation

Furong Zhang, Zhaonan Yang, Yulong Zhou, Binqiang Wang, Zhenming Xie, Ning Yu, Jie Zhao, Howard Goldfine, Shang Dai, Genlin Zhang, Bing Tian. Characterization and heterologous expression of plasmalogen synthase MeHAD from Megasphaera elsdenii. Biochimica et biophysica acta. Molecular and cell biology of lipids. 2023 Sep;1868(9):159358