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Title: Isolation and characterization of proteins involved in cryoprotection from cold resistant mutants of Pseudomonas fluorescens ATCC13525
Authors: Katiyar, Vandana
Mishra, D P
Kumar, Satish
Goel, Reeta
Keywords: cold resistant mutants;cryoprotection;freeze-labile enzyme;freeze survival;P. fluorescens
Issue Date: Jul-2004
Publisher: CSIR
IPC Code: Int. Cl.7 A 01 N 63/00
Abstract: The proteins isolated from cold resistant mutants of Pseudomonas fluorescens ATCC13525 were biologically characterized. These proteins, designated as CRP-1 (41.6 kDa, from CRPF9) and CRP-2 (14.1 kDa, from CRPF8), were able to protect freeze-labile alkaline phosphatase (ALP) against freeze denaturation up to approximately 84% activity. Addition of BSA could not yield similar cryoprotection of ALP. However, 60% loss of enzymatic activity was recorded in absence of the proteins. Furthermore, the two proteins in combination protected the enzyme activity up to 86%. The freeze-thaw challenge given to the mutants and wild type cells revealed the mutant to be more resistant to freezing (-20°C). The survival percentage of the mutant cells after repeated freeze thaw was significantly higher (55.6%) than wild type (9.8%). Nevertheless, incubation at 10°C, prior to freezing (-20°C) resulted in a further increase in survival percentage of mutant cells (79.6%). The proteins isolated from these mutants possessed the cryoprotective ability, which could be correlated with increased freezing resistance of the mutants at low temperature.
Page(s): 378-381
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Appears in Collections:IJBT Vol.03(3) [July 2004]

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