Gene cloning of Streptomyces phospholipase D P821 suitable for synthesis of phosphatidylserine

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Gene cloning of Streptomyces phospholipase D P821 suitable for synthesis of phosphatidylserine
Min-Woo Moon; Jung-Kee Lee; Tae Kwang Oh; C S Shin; H K Kim
Bibliographic Citation
Journal of Microbiology and Biotechnology, vol. 16, no. 3, pp. 408-413
Publication Year
A strain, P821, with phospholipase D activity was isolated from soil and identified as a Streptomyces species. The phospholipase D enzyme was purified from a culture broth of the isolated strain using ammonium sulfate precipitation and DEAE-Sepharose, phenyl-Sepharose, and Superose 12 HR column chromatographies. The purified enzyme exhibited an optimum temperature and pH of 55°C and 6.0, respectively, in the hydrolysis of phosphatidylcholine and remained stable up to 60°C within a pH range of 3.5-8.0. The enzyme also catalyzed a transphosphatidylation reaction to produce phosphatidylserine with phosphatidylcholine and serine substrates. The optimum conditions for the transphosphatidylation were 30°C and pH 5.0, indicating quite different optimum conditions for the hydrolysis and transphosphatidylation reactions. The gene encoding the enzyme was cloned by Southern hybridization and colony hybridization using a DNA probe designed from the conserved regions of other known phospholipase D enzymes. The resulting amino acid sequence was most similar to that of the PLD enzyme from Streptomyces halstedii (89.5%). Therefore, the enzyme was confirmed to be a phospholipase D with potential use in the production of phosphatidylserine.
phosphatidylserinephospholipase Dstreptomycestransphosphatidylation
Korea Soc-Assoc-Inst
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Division of Biomedical Research > Metabolic Regulation Research Center > 1. Journal Articles
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