Enhancement of secretion and extracellular stability of staphylokinase in Bacillus subtilis by wprA gene disruption.
Staphylokinase (SAK), a polypeptide secreted by Staphylococcus aureus, is a plasminogen activator with a therapeutic potential in thrombosis diseases. A Bacillus subtilis strain which is multiply deficient in exoproteases was transformed by an expression plasmid carrying a promoter and a signal sequence of subtilisin fused in frame with the sak open reading frame. However, the amount of SAK secretion was marginal (45 mg/liter). In contrast, disruption of the wprA gene, which encodes a subtilisin-type protease, strongly promoted the production of SAK in the stationary phase (181 mg/liter). In addition, the extracellular stability of mature SAK was dramatically enhanced. These data indicate a significant role of the wprA gene product in degrading foreign proteins, both during secretion and in the extracellular milieu.
Lee
S J
SJ
Department of Biological Sciences, Korea Advanced Institute of Science and Technology, Republic of Korea.
Kim
D M
DM
Bae
K H
KH
Byun
S M
SM
Chung
J H
JH
D001412:Bacillus subtilis / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D001426:Bacterial Proteins; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D017353:Gene Deletion; D015151:Immunoblotting; D008666:Metalloendopeptidases / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D012697:Serine Endopeptidases; C104015:WprA protein, Bacillus subtilis; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus