Mechanisms of heat inactivation of a proteinase from Pseudomonas fluorescens biotype I

Abstract

SummaryHeat inactivation of a metalloproteinase, isolated from Pseudomonas fluorescens biotype I strain 112, was investigated in the temperature ranges 50–60 °C and 90–140 °C. At 90 °C the denaturation of the enzyme followed first-order kinetics with a decimal reduction time of 110 min and a velocity constant K of 3·5 × 10−4 s−1. Activation energy Ea was 100 kJ/mol for this temperature range. In the 50–60 °C region the proteinase was inactivated by autolysis, as shown by electrophoresis and gel filtration. At 55 °C the decimal reduction time was ∼ 22 s, at 57 °C it was 8 s. Rapid inactivation at 55 °C was only possible if the enzyme was heated from lower temperatures, but not if cooled down from 90 °C. This is due to a conformational change of the protein at this temperature. A model for the description of heat inactivation in the two temperature ranges is proposed.