Extracellular beta-galactosidase activity of a Fibrobacter succinogenes S85 mutant able to catabolize lactose.
Fibrobacter succinogenes S85 is unable to grow with lactose as the source of carbohydrate, although it does exhibit low beta-galactosidase (EC 3.2.1.23) activity. Spontaneous mutants of strain S85 able to grow on lactose were isolated after spreading cells on a chemically defined agar medium with lactose as the carbohydrate source. A lactose-catabolizing isolate, designated L2, exhibited a sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profile and an immunoblot profile with polyclonal antibodies to whole cells of S85 which were identical to those observed for S85. Strain L2 exhibited both cell-associated and extracellular beta-galactosidase activity with either p-nitrophenyl-beta-D-galactopyranoside or lactose as the substrate. The cell-associated enzyme exhibited the greatest activity in the periplasmic space. Enzyme production was partially inhibited by glucose. The beta-galactosidase was activated by divalent cations and exhibited a pH optimum of 6.5. Analysis of the extracellular culture fluid revealed that glucose derived from the hydrolysis of lactose was used for growth, but galactose was not metabolized further. Cells were unable to take up the lactose analog, methyl-beta-D-thiogalactopyranoside. These data suggest that beta-galactosidase of F. succinogenes L2 cleaves lactose outside the cells and that the glucose released is catabolized while the galactose accumulates in the extracellular culture fluid.
Javorsky
P
P
Department of Microbiology, University of Guelph, Ontario, Canada.
Lee
S F
SF
Gibbins
A M
AM
Forsberg
C W
CW
D000818:Animals; D001421:Bacteria, Anaerobic / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D007785:Lactose / Q000378:metabolism*; D008858:Microscopy, Phase-Contrast; D009154:Mutation; D012417:Rumen / Q000382:microbiology*; D001616:beta-Galactosidase / Q000032:analysis / Q000378:metabolism*
D001616:beta-Galactosidase; D007785:Lactose