Study of the Bacillus subtilis ATCC21332 pbpE-racX operon in relation with the formation or disassembly of biofilms

Bacillus subtilis is a PGPR (Plant Growth Promoting Rhizobacterium) Gram positive bacterium and a model for studying the in vitro formation or disruption of biofilms. At the liquid/air interface of standing cultures, B. subtilis forms thick pellicles of limited lifetimes. Some D-amino acids have been reported among the factors playing a role in the disassembly of B. subtilis biofilms and ylmE or racX mutants (in which the racemases YlmE or RacX are absent) show a delay in pellicle disruption [I. Kolodkin et al. Science (2010) 328:627-629]. The racX encoding gene is part of a bicistronic operon in which the first gene (pbpE) codes for a Penicillin-Binding Protein, the PBP4* whose function is not characterized. Our studies aim to delete the complete pbpE-racX operon and compare the phenotypes of mutants and parental strains ATCC21332 or ATCC6051 in standing cultures. The substrate specificity of the purified RacX racemase is currently under investigation as well as the functional characterization of PBP4*, a protein possessing a lipocalin-like domain.