Properties of beta-galactosidase III: implications for entry of galactosides into Klebsiella

Abstract

Klebsiella sp. strain CT-200 lacks both its plasmid-borne lac operon, which specifies beta-galactosidase I, and its chromosomal lac operon, which specifies beta-galactosidase II, but it expresses a gene for a third beta-galactosidase, beta-galactosidase III, constitutively. CT-200 was examined to determine whether there was a beta-galactoside permease associated with the beta-galactosidase III gene. The failure of CT-200 to transport thiomethyl-beta-galactoside, o-nitrophenyl-beta-D-galactopyranoside, phenyl-beta-galactoside, lactulose, or galactosyl-arabinose was taken as evidence that beta-galactoside permease is not part of a beta-galactosidase III operon. Optimal assay conditions for beta-galactosidase II, whose activity was used as a measure of beta-galactoside transport, are reported here, as are an improved purification method and some physical and catalytic properties of the enzyme not previously reported.