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Asymmetric in vitro synthesis of diastereomeric phosphatidylglycerols from phosphatidylcholine and glycerol by bacterial phospholipase D
Author(s) -
Sato Rina,
Itabashi Yutaka,
Hatanaka Tadashi,
Kuksis Arnis
Publication year - 2004
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-004-1324-1
Subject(s) - glycerol , phospholipase d , diastereomer , stereochemistry , phosphatidylcholine , phosphatidic acid , chemistry , organic chemistry , biochemistry , phospholipid , enzyme , membrane
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.