Comparative studies of endonuclease I from cold-adapted Vibrio salmonicida and mesophilic Vibrio cholerae.
Endonuclease I is a periplasmic or extracellular enzyme present in many different Proteobacteria. The endA gene encoding endonuclease I from the psychrophilic and mildly halophilic bacterium Vibrio salmonicida and from the mesophilic brackish water bacterium Vibrio cholerae have been cloned, over-expressed in Escherichia coli, and purified. A comparison of the enzymatic properties shows large differences in NaCl requirements, optimum pH, temperature stability and catalytic efficiency of the two proteins. The V. salmonicida EndA shows typical cold-adapted features such as lower unfolding temperature, lower temperature optimum for activity, and higher specific activity than V. cholerae EndA. The thermodynamic activation parameters confirm the psychrophilic nature of V. salmonicida EndA with a much lower activation enthalpy. The optimal conditions for enzymatic activity coincide well with the corresponding optimal requirements for growth of the organisms, and the enzymes function predominantly as DNases at physiological concentrations of NaCl. The periplasmic or extracellular localization of the enzymes, which renders them constantly exposed to the outer environment of the cell, may explain this fine-tuning of biochemical properties.
Altermark
Bjørn
B
Norwegian Structural Biology Centre, Faculty of Science, University of Tromsø, Norway.
Niiranen
Laila
L
Willassen
Nils P
NP
Smalås
Arne O
AO
Moe
Elin
E
D044165:Aliivibrio salmonicida / Q000201:enzymology* / Q000502:physiology; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003080:Cold Temperature; D003850:Deoxyribonuclease I / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D004305:Dose-Response Relationship, Drug; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D012965:Sodium Chloride / Q000494:pharmacology; D013379:Substrate Specificity; D013696:Temperature; D014734:Vibrio cholerae / Q000201:enzymology* / Q000502:physiology
D012965:Sodium Chloride; D003850:Deoxyribonuclease I