A novel Ca2+-dependent phospholipase D from Streptomyces tendae, possessing only hydrolytic activity

  • Poonam Mander
  • Jaya Ram Simkhada
  • Seung Sik Cho
  • Sung Ju Park
  • Hong Seok Choi
  • Hei Chan Lee
  • Jae Kyung Sohng
  • Jin Cheol Yoo
Research Articles Drug Actions
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Abstract

An extracellular phospholipase D (PLD St ) was purified from Streptomyces tendae by two successive chromatographic steps on Sepharose CL-6B and DEAE-Sepharose CL-6B. Molecular weight of the PLD St was estimated to be approximately 43 kDa by sodium dodecyl sulfatepolyacrylamide gel electrophoresis. Maximal activity was at pH 8 and 60°C, and the enzyme was stable at or below 60°C and between pH 8 and 10, when assayed after 1.5 and 24 h, respectively. The enzyme activity had an absolute requirement of Ca2+, and the maximum activity was at 2 mM CaCl2. The Km and Vmax values for phosphatidyl choline were 0.95 mM and 810 µmol min−1 mg−1, respectively. More importantly, PLD St could not catalyze transphosphatidylation of glycerol, L-serine, myo-inositol and ethanolamine, which have been extensively used to evaluate the activity. The result strongly suggests that PLD St does not have the transphosphatidylation activity, thereby making it the first Streptomyces PLD possessing only hydrolytic activity. PLD St may therefore be a novel type of PLD enzyme.

Key words

Hydrolytic activity Novel phospholipase D Streptomyces tendae 
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Copyright information

© The Pharmaceutical Society of Korea and Springer Netherlands 2009

Authors and Affiliations

  • Poonam Mander
    • 1
  • Jaya Ram Simkhada
    • 1
  • Seung Sik Cho
    • 1
  • Sung Ju Park
    • 1
  • Hong Seok Choi
    • 1
  • Hei Chan Lee
    • 2
  • Jae Kyung Sohng
    • 2
  • Jin Cheol Yoo
    • 1
    • 3
  1. 1.Department of PharmacyChosun UniversityGwangjuKorea
  2. 2.Institute of Biomolecule ReconstructionSun Moon UniversityAsansi, ChungnamKorea
  3. 3.Department of Pharmacy, College of PharmacyChosun UniversityGwangjuKorea

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