Protein secretion in phosphate-limited cultures of Bacillus subtilis 168.
The secretion of proteins from Bacillus subtilis was studied under physiologically well-defined conditions in continuous cultures at a range of specific growth rates. The kinetics of secretion was analysed by using pulse-chase and immunoprecipitation techniques that allowed both processing and release to be monitored. Growth conditions were selected that were known to lead to significant changes in the anionic polymer composition of the cell wall. Under magnesium limitation only low levels of native proteins were released into the growth medium. In contrast, much higher amounts of released protein were observed under phosphate limitation. Although synthesis of native secretory proteins appeared to be highly regulated, only minor changes in the secretion of heterologous proteins were detected. Comparable kinetics of protein release of cells grown under different conditions indicated similar cell wall permeabilities. The large changes in the amounts of released proteins were not reflected in the production of chaperones and components required fro protein secretion. The data suggest that the capacity of the secretion machinery is not a major limiting step in the export of native secretory proteins.
Müller
J P
JP
Institut für Molekularbiologie, Friedrich-Schiller-Universität, Jena, Germany. jmueller@imb-jena.de
Harwood
C R
CR
D001412:Bacillus subtilis / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D008274:Magnesium / Q000494:pharmacology; D010710:Phosphates / Q000494:pharmacology*; D011994:Recombinant Proteins / Q000378:metabolism; D013997:Time Factors; D014169:Transformation, Bacterial / Q000235:genetics
D001426:Bacterial Proteins; D010710:Phosphates; D011994:Recombinant Proteins; D008274:Magnesium