Ovochymase in amphioxus Branchiostoma belcheri is an ovary-specific trypsin-like serine protease with an antibacterial activity.

Ovochymases have been shown to be present in vertebrates; little information is available at present regarding ovochymase in invertebrates. Here we isolated a cDNA encoding an ovochymase homolog from amphioxus Branchiostoma belcheri, named BbOvc. The cDNA contained a 1248bp open reading frame corresponding to a deduced protein of 415 amino acids with a predicted molecular mass of approximately 44.4kDa. Phylogenetic analysis showed that BbOvc was located at the base of its vertebrate counterparts, suggesting that it represents the archetype of vertebrate ovochymases. BbOvc is found to display a tissue- and stage-specific expression pattern, with a predominant expression in the ovary of sexually matured females and in the early stage embryos (1-16-cell embryos). The recombinant ovochymase expressed in vitro shows a trypsin-like activity capable of hydrolysing the trypsin prototypic substrate N(a)-benzoyl-l-arginine ethyl ester (60UBAEE/mg), which can be inhibited by the trypsin-specific inhibitor soybean trypsin inhibitor. It also exhibits an antibacterial activity capable of inhibiting the growth of bacteria like E. coli and V. parahaemolyticus. Taken together, these data indicate that BbOvc is a novel ovochymase with an antibacterial activity and offer first clues to its role as an immune-relevant molecule which may protect the early embryos from pathogenic attacks.