Native and heterologous protein secretion by Streptomyces lividans.
The secretion of the heterologous parathion phosphotriesterase in S. lividans using the Streptomyces beta-galactosidase signal sequence was further characterised using a pulse/chase system. Unsecreted cell-associated protein in both the precursor and signal-cleaved forms was observed when the protein was expressed from both low- and high-copy vectors. Fractionation of the cells followed by immunoprecipitation with phosphotriesterase antibody suggests that the precursor is membrane-bound while the signal cleaved form is present in the soluble fraction. Preliminary data on the processing of alpha-amylase, a native streptomyces protein, showed much more rapid processing and secretion, but nevertheless still revealed cell-associated, signal-cleaved protein.
Sathyamoorthy
M
M
Department of Pharmaceutical Sciences, School of Pharmacy, University of Maryland, Baltimore 21201, USA.
Stemke
D
D
Speedie
M K
MK
D000595:Amino Acid Sequence; D043303:Aryldialkylphosphatase; D002462:Cell Membrane / Q000201:enzymology; D004950:Esterases / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals; D011994:Recombinant Proteins / Q000378:metabolism; D017421:Sequence Analysis; D013302:Streptomyces / Q000201:enzymology*; D000516:alpha-Amylases / Q000378:metabolism
D011498:Protein Precursors; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D004950:Esterases; D043303:Aryldialkylphosphatase; D000516:alpha-Amylases