Tat dependent export of E. coli phytase AppA by using the PhoD-specific transport system of Bacillus subtilis.
It has been shown recently that the twin-arginine signal peptide of Bacillus subtilis phosphodiesterase PhoD (SPPhoD) can mediate Tat dependent transport of proteins via its specific Tat-transport components. In order to test the use of Tat dependent transport signals for heterologous product synthesis, Escherichia coli phytase AppA was expressed under control of PhoD-specific export signals in B. subtilis. Induction of Tat components TatAd/TatCd was mediated by using a functionally altered PhoR/PhoP signal transduction system which regulates the expression of these components. AppA was highly susceptible to host specific extracellular proteases. Expression of appA in B. subtilis wprA strain resulted in the stable production of AppA. A fusion protein consisting of SPPhoD and mature AppA remained unprocessed, while introduction of the AppA signal peptidase cleavage site resulted in efficient processing of the fusion protein.
Gerlach
Roman
R
Institute of Molecular Biology, Jena University, Hans-Knöll-Str. 2, D-07745 Jena, Germany.
Pop
Ovidiu
O
Müller
Jörg P
JP
D010832:6-Phytase / Q000235:genetics / Q000378:metabolism*; D000135:Acid Phosphatase / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D009154:Mutation; D010727:Phosphoric Diester Hydrolases / Q000235:genetics* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; C061279:PhoR protein, Bacteria; C061064:PhoP protein, Bacteria; D000135:Acid Phosphatase; D010832:6-Phytase; C406788:appA protein, E coli; D010727:Phosphoric Diester Hydrolases; D012697:Serine Endopeptidases; C104015:WprA protein, Bacillus subtilis