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Volume 146, Issue 1

Complete spore-cortex hydrolysis during germination of 168 requires SleB and YpeB Free

Abstract

The role of the gene of , which encodes a putative spore-cortex-lytic enzyme, and the downstream gene were investigated. Both SleB and YpeB were required for normal germination to occur. The corresponding mutants formed phase-bright, heat-resistant spores with no apparent defects in dormancy. However, mutant spore suspensions lost optical density slower than the wild-type and spores were phase-grey even 12 h after the triggering of germination. Since the loss of heat resistance and release of dipicolinic acid was similar to the wild-type, these mutants were blocked in the later stages of germination. The mutants were nevertheless capable of outgrowth on rich agar to form colonies, indicating that other spore components can compensate for their function sufficiently to allow outgrowth. The expression and regulation of the operon was examined using a transcriptional fusion. Expression of the operon began 2 h after the onset of sporulation and was under the control of RNA polymerase containing the forespore-specific sigma factor, σ. The application of reverse phase HPLC revealed that the mutants do not have any structural defect in the dormant spore cortex and therefore these genes are not required for normal spore-cortex synthesis. The analysis of peptidoglycan dynamics during germination showed, however, that the cortex was only partially hydrolysed in both mutants. This analysis also revealed that the likely hydrolytic bond specificity of SleB is likely to be that of a lytic transglycosylase.

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  • Accepted:
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  • Published Online:
Keyword(s): Bacillus subtilis , cortex hydrolysis , endospores , germination , GSLE, germination-specific lytic enzyme , MUG, methylumbelliferyl β-d-galactoside , peptidoglycan and RP-HPLC, reverse phase HPLC
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2000-01-01
2024-04-20
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Complete spore-cortex hydrolysis during germination of Bacillus subtilis 168 requires SleB and YpeB
Microbiology 146, 57 (2000); https://doi.org/10.1099/00221287-146-1-57
/content/journal/micro/10.1099/00221287-146-1-57
/content/journal/micro/10.1099/00221287-146-1-57
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