In the exponentially growing vegetative cells of Allomyces arbuscula we have identified three more enzyme activities using synthetic peptide substrates. These were in addition to two identified previously as Ca²⁺-dependent proteases (CDP I and CDP II) cleaving peptides at Arg in primary cleavage site (P1 position). One of the three new enzymes specifically cleaved peptides at Arg, another at Ala. Leu or Phe and the third prefered Tyr in P1 position. All these activities were Ca²⁺-independent. The activity showing preference for Ala in P1 position was different from the similar enzyme from differentiating cells since it was not inhibited by the serine protease inhibitors at concentrations known to inhibit differentiating cell enzyme or trypsin, both serine proteases. These enzymes were not inhibited by classical protease inhibitors.