Article
Removal of a proteolytic activity associated with aggregates formed from expression of creatine kinase in Escherichia coli leads to improved recovery of active enzyme.
Bio/technology (Nature Publishing Company) - 1 Oct 1990
Babbitt P C, West B L, Buechter D D, Kuntz I D, Kenyon G L
Abstract excerpt
Expression of creatine kinase (CK) from a Torpedo californica electric organ cDNA in Escherichia coli results in an insoluble protein product with no detectable CK activity. Although this is a stable aggregate that can be isolated in an enriched form by centrifugation, initial attempts to generate enzyme activity by denaturing and refolding yielded only minute amounts of active protein. We find that these low...
Topics
- Animals
- Aprotinin
- Cloning, Molecular
- Creatine Kinase
- Electrophoresis, Polyacrylamide Gel
- Enzyme Activation
- Escherichia coli
- Glucosides
- Inclusion Bodies
- Mutation