The effect of signal sequences on the efficiency of secretion of a heterologous phosphotriesterase by Streptomyces lividans.
A heterologous phosphotriesterase (parathion hydrolase) containing the native Flavobacterium species signal sequence was previously shown to be secreted by Streptomyces lividans. Western blot analysis of the recombinant phosphotriesterase produced by S. lividans demonstrated only the mature form extracellularly but both processed and unprocessed forms in cell-associated samples. To investigate the efficiency of secretion in Streptomyces, a construction was made that substituted a native Streptomyces beta-galactosidase signal sequence for the Flavobacterium signal sequence. This resulted in a higher proportion of hydrolase in the extracellular fluid and a lower proportion of parathion hydrolase remaining cell-associated. These results suggest that use of a native Streptomyces signal sequence may result in more efficient secretion of heterologous proteins.
Department of Biomedicinal Chemistry, School of Pharmacy, University of Maryland, Baltimore 21201.
S S
SS
Rowland
J J
JJ
Zulty
M
M
Sathyamoorthy
B M
BM
Pogell
M K
MK
Speedie
D000595:Amino Acid Sequence; D043303:Aryldialkylphosphatase; D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D005417:Flavobacterium / Q000201:enzymology; D008969:Molecular Sequence Data; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D021382:Protein Sorting Signals / Q000235:genetics / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000378:metabolism*; D013302:Streptomyces / Q000502:physiology*; D001616:beta-Galactosidase / Q000235:genetics
D001426:Bacterial Proteins; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D010744:Phosphoric Monoester Hydrolases; D043303:Aryldialkylphosphatase; D001616:beta-Galactosidase