ThermoSlope: A Software for Determining Thermodynamic Parameters from Single Steady-State Experiments.

ABSTRACT: The determination of the temperature dependence of enzyme catalysis has traditionally been a labourious undertaking. We have developed a new approach to the classical Arrhenius parameter estimation by fitting the change in velocity under a gradual change in temperature. The evaluation with a simulated dataset shows that the approach is valid. The approach is demonstrated as a useful tool by characterizing the Bacillus pumilus LipA enzyme. Our results for the lipase show that the enzyme is psychrotolerant, with an activation energy of 15.3 kcal/mol for the chromogenic substrate para-nitrophenyl butyrate. Our results demonstrate that this can produce equivalent curves to the traditional approach while requiring significantly less sample, labour and time. Our method is further validate

SUBMITTER: Lund BA 

PROVIDER: S-EPMC8658824 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature