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Applied and Environmental Microbiology, February 2000, p. 476-480, Vol. 66, No. 2
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Enhancement of Secretion and Extracellular Stability of Staphylokinase in Bacillus subtilis by wprA Gene Disruption

Sang Jun Lee,1 Dong Min Kim,1 Kwang Hee Bae,1 Si Myung Byun,1,2 and Jae Hoon Chung1,*

Department of Biological Sciences, Korea Advanced Institute of Science and Technology,1 and Research Center for New Bio-Materials in Agriculture,2 Taejon, 305-701, Republic of Korea

Received 7 July 1999/Accepted 3 November 1999

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.


* Corresponding author. Mailing address: Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Taejon 305-701, Korea. Phone: 82-42-869-2631. Fax: 82-42-869-2610. E-mail: jhchung{at}sorak.kaist.ac.kr.


Applied and Environmental Microbiology, February 2000, p. 476-480, Vol. 66, No. 2
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.



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