Archives of Microbiology

, Volume 156, Issue 3, pp 186–191 | Cite as

Intracellular serine protease-4, a new intracellular serine protease activity from Bacillus subtilis

  • Shannon M. Sheehan
  • Robert L. Switzer
Original Papers


A previously undiscovered intracellular serine protease activity, which we have called intracellular serine protease-4, was identified in extracts of stationary Bacillus subtilis cells, purified 260 fold from the cytoplasmic fraction, and characterized. The new protease was stable and active in the absence of Ca2+ ions and hydrolyzed azocasein and the chromogenic substrate carbobenzoxy-carbonyl-alanyl-alanyl-leucyl-p-nitroanilide, but not azocollagen or a variety of other chromogenic substrates. The protease was strongly inhibited by phenylmethylsulfonylfluoride, chymostatin and antipain, but not by chelators, sulfhydryl-reactive agents or trypsin inhibitors. Its activity was stimulated by Ca2+ ions and gramicidin S; its pH and temperature optima were 9.0 and 37°C, respectively. Although intracellular serine protease-4 was immunochemically distinct from intracellular serine protease-1, it was absent from a mutant in which the gene encoding the latter was disrupted.

Key words

Serine protease Bacillus subtilis Protease specificity Enzyme characterization 


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Copyright information

© Springer-Verlag 1991

Authors and Affiliations

  • Shannon M. Sheehan
    • 1
  • Robert L. Switzer
    • 1
  1. 1.Department of BiochemistryUniversity of IllinoisUrbanaUSA

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