Thermal inactivation and reactivation of an enzyme in vivo. Pantothenate hydrolase of Pseudomonas fluorescens.
Thermal inactivation and reactivation of pantothenate hydrolase were studied in whole cells of Pseudomonas fluorescens. The enzyme is susceptible to thermal inactivation in whole cells at 37-40 degrees C, and is reactivated when the temperature is lowered again. Chloramphenicol does not prevent reactivation. The activation energy of enzyme inactivation in vivo is about 540kJ/mol. This activation energy is 220kJ/mol in vitro, but it is increased to 550-630kJ/mol by several metabolites, such as succinate, glyoxylate and oxalate. Generally, good carbon sources, causing rapid growth, protect the enzyme from thermal inactivation in vivo, and enable reactivation to occur at a fast rate. The enzyme is also inactivated below 35 degrees C, showing an activation energy of about 35kJ/mol. Good carbon sources prevent this inactivation as well, and cause slight reactivation. Glycine, although not utilized for growth, protects the enzyme well from this inactivation but not from inactivation at 37-40 degrees C, and prevents reactivation totally. From the activation energies of inactivation and the effects of the various carbon sources, it appears possible that changes in the concentrations of intracellular metabolites may be responsible for the changes in inactivation and reactivation.
D002247:Carbon Isotopes; D002701:Chloramphenicol; D004789:Enzyme Activation; D004983:Ethanolamines; D005947:Glucose; D005971:Glutamates; D005998:Glycine; D006038:Glyoxylates; D006358:Hot Temperature; D006867:Hydrolases; D008715:Methionine / Q000378:metabolism; D010070:Oxalates; D010205:Pantothenic Acid; D011422:Propionates; D011549:Pseudomonas / Q000201:enzymology*; D011551:Pseudomonas fluorescens / Q000201:enzymology; D013386:Succinates
D002247:Carbon Isotopes; D004983:Ethanolamines; D005971:Glutamates; D006038:Glyoxylates; D010070:Oxalates; D011422:Propionates; D013386:Succinates; D010205:Pantothenic Acid; D002701:Chloramphenicol; D008715:Methionine; D006867:Hydrolases; D005947:Glucose; D005998:Glycine