Newly induced beta-galactosidase molecules have a higher activity than the basally expressed enzyme.
OBJECTIVE
The objective of this study was to compare the activities of individual molecules of induced and basally expressed Escherichia coli beta-galactosidase.


BACKGROUND
Single-molecule assays of enzymes have determined that individual molecules are not identical. They differ with respect to catalytic rate. The structural cause and cellular role of this microheterogeneity is as yet unknown.


METHODS
E. coli were grown and induced to produce beta-galactosidase by treatment with isopropyl-beta-D-thiogalactopyranoside. Cells were lysed and the beta-galactosidase assayed with capillary electrophoresis instrumentation utilizing post-column, laser-induced fluorescence detection. The enzyme obtained from treated cells were compared to that from untreated cells.


RESULTS
The activity of newly induced beta-galactosidase was found to be approximately 20% greater than that of the basally expressed enzyme. This measured difference is statistically significant.


CONCLUSIONS
Production of beta-galactosidase in E. coli under differing conditions results in differences in the activities of the individual enzyme molecules.
Craig
D B
DB
Chemistry Department, University of Winnipeg, Manitoba, Canada. dcraig@uwinnipeg.ca
Hall
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T
D019075:Electrophoresis, Capillary; D004790:Enzyme Induction; D004926:Escherichia coli / Q000201:enzymology*; D005453:Fluorescence; D007834:Lasers; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000378:metabolism*
D001616:beta-Galactosidase