microbiology, Vol 140, 1403-1410, Copyright © 1994 by Society for General Microbiology

ARTICLES

A spore-lytic enzyme released from Bacillus cereus spores during germination

S Makino, N Ito, T Inoue, S Miyata and R Moriyama
Department of Applied Biological Science, School of Agricultural Science, Nagoya University, Aichi, Japan.

The exudate of fully germinated spores of Bacillus cereus IFO 13597 in 0.25 M sodium phosphate buffer, pH 7.0, was found to contain a spore- lytic enzyme. This enzyme was found to cause loss of absorbance in coat- stripped spore suspensions and phase-darkening of the spores but had minimal activity on isolated peptidoglycan substrates. The enzyme was purified in an active form and identified as a 24 kDa protein which is either an amidase or a peptidase. The amino-terminal 19 residues had the following sequence: FSNQVIQRGASGEKVIELQ. The spore-lytic enzyme retained its activity in a medium of a relatively high ionic strength containing a non-ionic surfactant such as nonaethyleneglycol n-dodecyl ether. This activity was optimum at a salt concentration of about 30 mM in assay buffer at neutral pH. In contrast to the enzyme in a spore- bound form, the enzyme in solution was shown to be heat-sensitive and was readily inactivated by thiol reagents.

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