1887

Abstract

Effects of -alanine racemase, D-alanine, heat-activation and ammonia on germination of T spores were studied. Michaelis-Menten parameters for -alanine racemase activity in intact spores were: (-alanine)=3·6 m; (-alanine) = 1·2 m; (- to -alanine) = 1·6 μmol min (mg dry spores); (- to -alanine) = 0·53 μmol min (mg dry spores). Racemase activity was irreversibly inhibited with pseudo first order kinetics by incubating intact spores with 10 m--cycloserine. (Aminooxy)acetate (50 μ) completely inhibited racemase activity with 100 m--alanine as substrate. -Cysteine was a moderately effective racemase inhibitor. Comparisons of the effects of racemase inhibitors, -alanine and spore concentration on germination rates in 1 m--alanine indicated that racemization of -alanine accounted for nearly total inhibition of germination at spore concentrations exceeding about 50 μg ml, in the range conventionally used for spectrophotometric germination assays. Heat-activation decreased the inhibitory effect of -alanine, increasing the germination rate in 2 m-racemic alanine by a factor of 26. Using spores with an inhibited alanine racemase, germination rates in 1 m--alanine were stimulated by 40 m-NHCl to an extent comparable to the stimulation obtained by heat-activating the spores. Germination of unactivated spores in 1 m--alanine + 40 m-NHCl was completely inhibited by 1 m--alanine. These results clarified the roles of -alanine and -alanine racemase in controlling the germination of unactivated spores and suggested that the germinative functions of ammonia and the stereoisomers of alanine may be related.

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/content/journal/micro/10.1099/00221287-130-12-3123
1984-12-01
2021-07-26
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