Inducible production and cellular location of the epidermin biosynthetic enzyme EpiB using an improved staphylococcal expression system.
The antimicrobial peptide epidermin is distinguished by thioether amino acids such as meso-lanthionine, 3-methyl-lanthionine, and 2-aminovinylcysteine. The enzyme EpiB, encoded on a plasmid of the producing strain Staphylococcus epidermidis Tü3298, is very likely involved in the formation of these unusual amino acids. In order to obtain high-level production of EpiB, an improved staphylococcal expression vector based on the xylose-inducible xylA promoter of Staphylococcus xylosus was constructed. As shown by the expression of a lipase reporter gene, the new plasmid pTX15 mediated a considerably higher expression level after induction and a lower background expression level in the uninduced state than the previously described vector pCX15. The epiB gene was inserted in pTX15 and expressed in Staphylococcus carnosus. The EpiB protein was detected both in the cytoplasmic and the membrane fraction and was partially purified in three steps.
Peschel
A
A
Universität Tübingen, Germany.
Ottenwälder
B
B
Götz
F
F
D000900:Anti-Bacterial Agents / Q000096:biosynthesis*; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D001430:Bacteriocins; D001483:Base Sequence; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D010455:Peptides; D013210:Staphylococcus / Q000235:genetics* / Q000378:metabolism*; D013212:Staphylococcus epidermidis / Q000235:genetics / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D001430:Bacteriocins; D017931:DNA Primers; D004269:DNA, Bacterial; C085950:EpiB protein, Staphylococcus epidermidis; D008565:Membrane Proteins; D010455:Peptides; C050558:epidermin