The Bacillus subtilis system was developed as a host for gram-positive bacteria in a variety of applications, including agricultural, medical and food biotechnology and the production of recombinant proteins.
The Bacillus subtilis system is advantageous in that it is non-pathogenic, does not have significant bias in codon usage, and is capable of secreting functional extracellular proteins directly into the culture medium. A large body of information concerning transcription, translation, protein folding and secretion mechanisms, genetic manipulation, and large-scale fermentation is available.
The Bacillus Food Grade Expression System was created to make the advantages of the Bacillus expression system also accessible to areas in which antibiotic resistance gene markers are prohibited, such as the food and feed industry. It enables stable vector-based large scale heterologous protein production by an alternative selection, without antibiotics.
The Bacillus Food Grade Selection System is based on the interplay of an endogenous Bacillustoxin EndoA (expressed from vector) and its antitoxin EndoB (expressed from genome).
Two vectors are available for this system, differing in their origin of replication and copy number concerning B. subtilis:
For easy handling, the vectors are designed as B. subtilis / E. coli shuttle vectors. The parts of the vector used for cloning with E. coli can be eliminated afterwards by restriction enzyme cleavage and religation of the vector. This technique connects the advantage of easy cloning (with E. coli) with the food grade property of B. subtilis.
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