Development of an inducible and enhancible expression and secretion system in Bacillus subtilis

A set of inducible secretion vectors has been developed in Bacillus subtilis based on the regulatory region and the signal peptide sequence of the sacB gene encoding an extracellular enzyme, levansucrase. The expression of the inserted foreign gene (bla) encoding TEM beta-lactamase (Bla), can be induced by the addition of sucrose to the medium. Either the installation of a sacQ expression cassette into the same secretion vector, or the use of a sacUh two-protease-deficient strain (WB30), can significantly enhance expression of the bla gene. However, the combined use of the sacQ-containing secretion vector and the WB30 strain results in no further increase in Bla activity. During development of the secretion vector, the nucleotide sequence around the signal peptidase cleavage site has been redesigned, so that unique restriction sites were installed to facilitate the insertion of foreign genes.