Applied Biochemistry and Biotechnology

, Volume 168, Issue 2, pp 327–338 | Cite as

Characterization of a New Cold-adapted Lipase from Pseudomonas sp. TK-3

  • Daisuke Tanaka
  • Satoru Yoneda
  • Yoko Yamashiro
  • Akihiro Sakatoku
  • Takuro Kayashima
  • Kasumi Yamakawa
  • Shogo Nakamura
Article
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Accepted:

Abstract

A psychrotrophic Pseudomonas sp. TK-3 was isolated from dirty and cool stream water in Toyama, Japan from which we cloned and characterized the bacterial lipase LipTK-3. The sequenced DNA fragment contains an open reading frame of 1,428 bp that encoded a protein of 476 amino acids with an estimated molecular mass of 50,132 Da. The lipase showed high sequence similarity to those of subfamily Ι.3 lipase and had a conserved GXSXG motif around the catalytic Ser residue. Its optimal temperature was 20–25 °C, lower than in most other subfamily Ι.3 lipases. The lipase exhibited about 30 % of maximal activity at 5 °C. The optimal pH value was 8.0. The activity was strongly inhibited by EDTA and was highly dependent on Ca2+. Tricaprylin and p-nitrophenyl caprylate were the most favorable substrates among the triglycerides and p-nitrophenyl esters, respectively. LipTK-3 also showed high activity towards natural substrates including edible vegetable oils and animal fats. Furthermore, LipTK-3 was very active and stable in the presence of several detergents, metal ions, and organic solvents. This cold-adapted lipase may prove useful for future applications.

Keywords

Pseudomonas sp. Cold-adapted lipase Characterization 
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Notes

Acknowledgements

We thank Dr. S. I. Aizawa of the Prefectural University of Hiroshima for assistance with transmission electron microscopy.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Daisuke Tanaka
    • 1
  • Satoru Yoneda
    • 1
  • Yoko Yamashiro
    • 1
  • Akihiro Sakatoku
    • 1
  • Takuro Kayashima
    • 1
  • Kasumi Yamakawa
    • 1
  • Shogo Nakamura
    • 1
  1. 1.Graduate School of Science and EngineeringUniversity of ToyamaToyamaJapan

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