Heterologous expression of nonribosomal peptide synthetases in B. subtilis: construction of a bi-functional B subtilis/E coli shuttle vector system.
A major obstacle in investigating the biosynthesis of pharmacologically important peptide antibiotics is the heterologous expression of the giant biosynthetic genes. Recently, the genetically engineered strain Bacillus subtilis KE30 has been reported as an excellent surrogate host for the heterologous expression of an entire nonribosomal peptide synthetase (NRPS) gene cluster. In this study, we expand the applicability of this strain, by the development of four Escherichia coli/B. subtilis shuttle expression vectors. Comparative overproduction of hybrid NRPS proteins derived from both organisms revealed a significant beneficial effect of overproducing proteins in B. subtilis KE30 as underlined by the production of stable nondegradative proteins, as well as the formation of active phosphopantetheinylated holo-proteins.
Doekel
Sascha
S
Philipps-Universität Marburg, Fachbereich Chemie/Biochemie, Hans-Meerwein-Strasse, 35032, Marburg, Germany.
Eppelmann
Katrin
K
Marahiel
Mohamed A
MA
D000900:Anti-Bacterial Agents / Q000096:biosynthesis; D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics*; D001435:Bacteriophages / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005798:Genes, Bacterial; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics*; D016680:Genome, Bacterial; D010453:Peptide Synthases / Q000096:biosynthesis* / Q000235:genetics; D010455:Peptides; D011401:Promoter Regions, Genetic; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics; D014170:Transformation, Genetic
D000900:Anti-Bacterial Agents; D010455:Peptides; D011994:Recombinant Proteins; D010453:Peptide Synthases