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Molecular Biology Reports

, 37:939 | Cite as

Molecular cloning and characterization of phospholipase D from Jatropha curcas

  • Bin Liu
  • Lin Yao
  • Wenguo Wang
  • Jihai Gao
  • Fang Chen
  • Shenghua Wang
  • Ying Xu
  • Lin Tang
  • Yongjiong Jia
Article

Abstract

Phospholipase D (PLD, EC 3.1.4.4) is a key enzyme involved in phospholipid catabolism, initiating a lipolytic cascade in membrane deterioration during senescence and stress, which was cloned from Jatropha curcas L., an important plant species as its seed is the raw material for biodiesels. The cDNA was 2,886 bp in length with a complete open reading frame of 2,427 bp which encoded a polypeptide of 808 amino acids including a putative signal peptide of 53 amino acid residues and a mature protein of 755 amino acids with a predicted molecular mass of 86 kD and a pI of 5.44, having two highly conserved ‘HKD’ motifs. Phylogenetic analysis indicated the J. curcas PLD alpha (JcPLDα) showed a high similarity to other PLD alpha from plants. Semi-quantitative RT-PCR analysis revealed that it was especially abundant in root, stem, leaf, endosperm and flower, weakly in seed. And the JcPLDα was increasedly expressed in leaf undergoing environmental stress such as salt (300 mM NaCl), drought (30% PEG), cold (4°C) and heat (50°C). The JcPLDα protein was successfully expressed in Escherichia coli and showed high enzymatic activities. Maximal activity was at pH 8 and 60°C.

Keywords

Phospholipase D Jatropha curcas Molecular cloning Expression Enzyme activity 

Notes

Acknowledgments

We thank the reviewers for useful suggestions. This research is supported by grants from “11th Five-Year Plan” to support science and technology project (no. 2007BAD50B05) and Ministry of education key program (no. 307023).

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Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • Bin Liu
    • 1
  • Lin Yao
    • 2
  • Wenguo Wang
    • 1
  • Jihai Gao
    • 1
  • Fang Chen
    • 1
  • Shenghua Wang
    • 1
  • Ying Xu
    • 1
  • Lin Tang
    • 1
  • Yongjiong Jia
    • 1
  1. 1.College of Life SciencesSichuan UniversityChengduPeople’s Republic of China
  2. 2.Institute of Plant ProtectionSichuan Academy of AgriculturalChengduPeople’s Republic of China

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