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Separation of Two Functional Roles of L-Alanine in the Initiation of Bacillus subtilis Spore Germination

¹ Laboratory of Molecular Biology, National Institute of Neurological Diseases and Blindness, Public Health Service, Bethesda, Maryland 20014

ABSTRACT

Spores of the standard transformable Marburg strain of Bacillus subtilis can be initiated to germinate by L-alanine alone. We isolated mutants which required for this process, in addition to L-alanine, the combination of D-glucose + D-fructose + K⁺ or NH₄⁺ ions. In place of fructose, autoclaved or caramelized glucose could be used. Even the standard type strain required the addition of these three agents when D-alanine was present or when the temperature was raised. These findings show that L-alanine normally performs two functions during initiation, one of which is absent in the mutants or is blocked by D-alanine or elevated temperature. One of our mutants was not absolutely dependent on the addition of external L-alanine, because it could be initiated at a reduced rate by the sole addition of glucose + K⁺ or NH₄⁺. When K⁺ or NH₄⁺ was replaced by Na⁺, the initiation rate was greatly reduced. The divalent metal ions Mg⁺⁺, Mn⁺⁺, and Ca⁺⁺ could not satisfy the cation requirement.

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