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Asymmetric in vitro synthesis of diastereomeric phosphatidylglycerols from phosphatidylcholine and glycerol by bacterial phospholipase D

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Lipids

Abstract

Using chiral-phase HPLC, we determined the stereochemical configuration of the phosphatidylglycerols (PtdGro) synthesized in vitro from 1,2-diacyl-sn-glycero-3-phosphocholine (PtdCho, R configuration) or 1,2-diacyl-sn-glycero-3-phosphoethanolamine (PtdEtn, R configuration) and glycerol by transphosphatidylation with bacterial phospholipase D (PLD). The results obtained with PLD preparations from three Streptomyces strains (S. septatus TH-2, S. halstedii K5, and S. halstedii subsp. scabies K6) and one Actinomadura species were compared with those obtained using cabbage and peanut PLD. The reaction was carried out at 30°C in a biphasic system consisting of diethyl ether and acetate buffer. The resulting PtdGro were then converted into bis(3,5-dinitrophenylurethane) derivatives, which were separated on an (R)-1-(1-naphthyl)ethylamine polymer. In contrast to the cabbage and peanut PLD, which gave equimolar mixtures of the R,S and R,R diastereomers, as previously established, the bacterial PLD yielded diastereomixtures of 30–40% 1,2-diacyl-sn-glycero-3-phospho-1′-sn-glycerol (R,S configuration) and 60–70% 1,2-diacyl-sn-glycero-3-phospho-3′-sn-glycerol (R,R configuration). The highest disproportionation was found for the Streptomyces K6 species. The present study demonstrates that bacterial PLD-catalyzed transphosphatidylation proceeds to a considerable extent stereoselectively to produce PtdGro from PtdCho or PtdEtn and prochiral glycerol, indicating a preference for the sn-3′ position of the glycerol molecule.

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Abbreviations

DNPU:

3,5-dinitrophenylurethane

GroPCho:

glycerophosphocholine

GroPEtn:

glycerophosphoethanolamine

sn-3,sn-3′ (R,R):

1,2-diacyl-sn-glycero-3-phospho-3′-sn-glycerol

sn-3,sn-1′ (R,S):

1,2-diacyl-sn-glycero-3-phospho-1′-sn-glycerol

GroPGro:

glycerophosphoglycerol

PLD:

phospholipase D

PtdCho:

phosphatidylcholine

PtdEtn:

phosphatidylethanolamine

PtdGro:

phosphatidylglycerol

PtdOH:

phosphatidic acid

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Authors and Affiliations

  1. Research Institute for Biological Sciences, 716-1241, Okayama, Japan

    Tadashi Hatanaka

  2. Banting and Best Department of Medical Research, University of Toronto, M5G 1L6, Toronto, Ontario, Canada

    Arnis Kuksis

  3. Laboratory of Bioresources Chemistry, Graduate School of Fisheries Sciences, Hokkaido University, 3-1-1 Minato-cho, Hakodate, 041-8611, Hokkaido, Japan

    Rina Sato & Yutaka Itabashi

Authors
  1. Rina Sato
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  2. Yutaka Itabashi
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  3. Tadashi Hatanaka
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  4. Arnis Kuksis
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Correspondence to Yutaka Itabashi.

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Sato, R., Itabashi, Y., Hatanaka, T. et al. Asymmetric in vitro synthesis of diastereomeric phosphatidylglycerols from phosphatidylcholine and glycerol by bacterial phospholipase D. Lipids 39, 1013–1018 (2004). https://doi.org/10.1007/s11745-004-1324-1

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  • DOI: https://doi.org/10.1007/s11745-004-1324-1

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