Current Microbiology

, Volume 43, Issue 5, pp 365–370 | Cite as

Relationship Between Antifreeze Protein and Freezing Resistance in Pseudomonas putida GR12-2

  • Hidehisa Kawahara
  • Jiping Li
  • Marilyn Griffith
  • Bernard R. Glick
Article

Abstract

Following transposon Tn5 mutagenesis of the plant growth-promoting rhizobacterium Pseudomonas putida GR12-2, mutants that have different freeze-resistant properties were selected. Five of the freeze-sensitive mutants, i.e. FSM-5, -6, -14, -29, and -41, secreted a lower amount of antifreeze protein-(AFP) into the culture broth compared with the wild-type. Among of these five mutants, the three mutants (FSM-6, FSM-14, and FSM-41) that have the lowest level of freezing resistance (4.0–6.0% survival) also produce AFP at low levels (0.5–0.9 μg/mL) compared with the wild-type (4.8 μg/ml). The antifreeze and ice-nucleating activities of the AFP from these three mutant strains were similar to those of wild-type. Furthermore, the decreased freezing resistance from three mutants could be partially restored by adding purified AFP to mutant cell suspensions. Freezing resistance of three mutants was found to increase in proportion to the addition of AFP up to a concentration of 50 μg/mL. We conclude that accumulation of AFP is one component of the mechanism for freezing resistance in bacteria.

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Copyright information

© Springer-Verlag New York Inc. 2001

Authors and Affiliations

  • Hidehisa Kawahara
    • 1
  • Jiping Li
    • 2
  • Marilyn Griffith
    • 2
  • Bernard R. Glick
    • 2
  1. 1.Department of Biotechnology, Kansai University, Suita, Osaka, 564-8680Japan
  2. 2.Department of Biology, University of Waterloo, Waterloo, Ontario, N2L 3G1Canada

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