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 sequ...
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| Veröffentlicht in: | Applied and environmental microbiology 2000-02, Vol.66 (2), p.476-480 |
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| creator | Lee, S J Kim, D M Bae, K H Byun, S M Chung, J H |
| description | 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. |
| doi_str_mv | 10.1128/AEM.66.2.476-480.2000 |
| format | Article |
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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.</description><identifier>ISSN: 0099-2240</identifier><identifier>EISSN: 1098-5336</identifier><identifier>DOI: 10.1128/AEM.66.2.476-480.2000</identifier><identifier>PMID: 10653706</identifier><identifier>CODEN: AEMIDF</identifier><language>eng</language><publisher>Washington, DC: American Society for Microbiology</publisher><subject>Amino acids ; Bacillus subtilis ; Bacillus subtilis - enzymology ; Bacillus subtilis - genetics ; Bacillus subtilis - growth & development ; Bacteria ; Bacterial Proteins ; Biological and medical sciences ; Biotechnology ; Electrophoresis, Polyacrylamide Gel ; Enzyme Stability ; Enzymes ; Fundamental and applied biological sciences. Psychology ; Gene Deletion ; Genes ; Genetics and Molecular Biology ; Health. Pharmaceutical industry ; Immunoblotting ; Industrial applications and implications. Economical aspects ; Metalloendopeptidases - genetics ; Metalloendopeptidases - metabolism ; Molecular biology ; Other active biomolecules ; Peptides ; Plasmids - genetics ; Production of active biomolecules ; Proteins ; Recombinant Proteins - metabolism ; sak gene ; SAK protein ; Serine Endopeptidases - genetics ; Serine Endopeptidases - metabolism ; Staphylococcus aureus ; staphylokinase ; Transformation, Bacterial ; wprA gene</subject><ispartof>Applied and environmental microbiology, 2000-02, Vol.66 (2), p.476-480</ispartof><rights>2000 INIST-CNRS</rights><rights>Copyright American Society for Microbiology Feb 2000</rights><rights>Copyright © 2000, American Society for Microbiology 2000</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c492t-b320a119a8eed85c55e877db9a582c6d6b977400d0eb6ef4a8ca1131aece039f3</citedby><cites>FETCH-LOGICAL-c492t-b320a119a8eed85c55e877db9a582c6d6b977400d0eb6ef4a8ca1131aece039f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC91851/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC91851/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,3188,27924,27925,53791,53793</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=1265495$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/10653706$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lee, S J</creatorcontrib><creatorcontrib>Kim, D M</creatorcontrib><creatorcontrib>Bae, K H</creatorcontrib><creatorcontrib>Byun, S M</creatorcontrib><creatorcontrib>Chung, J H</creatorcontrib><title>Enhancement of secretion and extracellular stability of staphylokinase in Bacillus subtilis by wprA gene disruption</title><title>Applied and environmental microbiology</title><addtitle>Appl Environ Microbiol</addtitle><description>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.</description><subject>Amino acids</subject><subject>Bacillus subtilis</subject><subject>Bacillus subtilis - enzymology</subject><subject>Bacillus subtilis - genetics</subject><subject>Bacillus subtilis - growth & development</subject><subject>Bacteria</subject><subject>Bacterial Proteins</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Enzyme Stability</subject><subject>Enzymes</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Gene Deletion</subject><subject>Genes</subject><subject>Genetics and Molecular Biology</subject><subject>Health. Pharmaceutical industry</subject><subject>Immunoblotting</subject><subject>Industrial applications and implications. Economical aspects</subject><subject>Metalloendopeptidases - genetics</subject><subject>Metalloendopeptidases - metabolism</subject><subject>Molecular biology</subject><subject>Other active biomolecules</subject><subject>Peptides</subject><subject>Plasmids - genetics</subject><subject>Production of active biomolecules</subject><subject>Proteins</subject><subject>Recombinant Proteins - metabolism</subject><subject>sak gene</subject><subject>SAK protein</subject><subject>Serine Endopeptidases - genetics</subject><subject>Serine Endopeptidases - metabolism</subject><subject>Staphylococcus aureus</subject><subject>staphylokinase</subject><subject>Transformation, Bacterial</subject><subject>wprA gene</subject><issn>0099-2240</issn><issn>1098-5336</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2000</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpdkU2P0zAQhi0EYsvCTwBZCHFLGTu2Y0tcyqp8SIu4wNlynMnWS-oEOwH673FpBQsnH_y8o3nnIeQpgzVjXL_abD-ulVrztWhUJTSsOQDcIysGRleyrtV9sgIwpuJcwAV5lPNtAQQo_ZBcMFCybkCtSN7GnYse9xhnOvY0o084hzFSFzuKP-fkPA7DMrhE8-zaMIT58Buc3bQ7DOPXEF1GGiJ943woZKZ5aefCZdoe6I8pbegNRqRdyGmZjqMfkwe9GzI-Ob-X5Mvb7eer99X1p3cfrjbXlReGz1Vbc3CMGacROy29lKibpmuNk5p71anWNI0A6ABbhb1w2he8Zg49Qm36-pK8Ps2dlnaPnS8VkxvslMLepYMdXbD__sSwszfjd2uYlqzEX57jafy2YJ7tPuTjMVzEccmWNcJIEKKAz_8Db8clxVLNcpBGSClUgeQJ8mnMOWH_Zw8G9mjUFqNWKcttMWqLUXs0WnLP7pa4kzopLMCLM-Cyd0Ofis6Q_3JcybJn_QuPRK0u</recordid><startdate>20000201</startdate><enddate>20000201</enddate><creator>Lee, S J</creator><creator>Kim, D M</creator><creator>Bae, K H</creator><creator>Byun, S M</creator><creator>Chung, J H</creator><general>American Society for Microbiology</general><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QO</scope><scope>7SN</scope><scope>7SS</scope><scope>7ST</scope><scope>7T7</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>SOI</scope><scope>5PM</scope></search><sort><creationdate>20000201</creationdate><title>Enhancement of secretion and extracellular stability of staphylokinase in Bacillus subtilis by wprA gene disruption</title><author>Lee, S J ; Kim, D M ; Bae, K H ; Byun, S M ; Chung, J H</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c492t-b320a119a8eed85c55e877db9a582c6d6b977400d0eb6ef4a8ca1131aece039f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2000</creationdate><topic>Amino acids</topic><topic>Bacillus subtilis</topic><topic>Bacillus subtilis - enzymology</topic><topic>Bacillus subtilis - genetics</topic><topic>Bacillus subtilis - growth & development</topic><topic>Bacteria</topic><topic>Bacterial Proteins</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Electrophoresis, Polyacrylamide Gel</topic><topic>Enzyme Stability</topic><topic>Enzymes</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Gene Deletion</topic><topic>Genes</topic><topic>Genetics and Molecular Biology</topic><topic>Health. Pharmaceutical industry</topic><topic>Immunoblotting</topic><topic>Industrial applications and implications. Economical aspects</topic><topic>Metalloendopeptidases - genetics</topic><topic>Metalloendopeptidases - metabolism</topic><topic>Molecular biology</topic><topic>Other active biomolecules</topic><topic>Peptides</topic><topic>Plasmids - genetics</topic><topic>Production of active biomolecules</topic><topic>Proteins</topic><topic>Recombinant Proteins - metabolism</topic><topic>sak gene</topic><topic>SAK protein</topic><topic>Serine Endopeptidases - genetics</topic><topic>Serine Endopeptidases - metabolism</topic><topic>Staphylococcus aureus</topic><topic>staphylokinase</topic><topic>Transformation, Bacterial</topic><topic>wprA gene</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lee, S J</creatorcontrib><creatorcontrib>Kim, D M</creatorcontrib><creatorcontrib>Bae, K H</creatorcontrib><creatorcontrib>Byun, S M</creatorcontrib><creatorcontrib>Chung, J H</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Environment Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>Environment Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Applied and environmental microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lee, S J</au><au>Kim, D M</au><au>Bae, K H</au><au>Byun, S M</au><au>Chung, J H</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Enhancement of secretion and extracellular stability of staphylokinase in Bacillus subtilis by wprA gene disruption</atitle><jtitle>Applied and environmental microbiology</jtitle><addtitle>Appl Environ Microbiol</addtitle><date>2000-02-01</date><risdate>2000</risdate><volume>66</volume><issue>2</issue><spage>476</spage><epage>480</epage><pages>476-480</pages><issn>0099-2240</issn><eissn>1098-5336</eissn><coden>AEMIDF</coden><abstract>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.</abstract><cop>Washington, DC</cop><pub>American Society for Microbiology</pub><pmid>10653706</pmid><doi>10.1128/AEM.66.2.476-480.2000</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Amino acids Bacillus subtilis Bacillus subtilis - enzymology Bacillus subtilis - genetics Bacillus subtilis - growth & development Bacteria Bacterial Proteins Biological and medical sciences Biotechnology Electrophoresis, Polyacrylamide Gel Enzyme Stability Enzymes Fundamental and applied biological sciences. Psychology Gene Deletion Genes Genetics and Molecular Biology Health. Pharmaceutical industry Immunoblotting Industrial applications and implications. Economical aspects Metalloendopeptidases - genetics Metalloendopeptidases - metabolism Molecular biology Other active biomolecules Peptides Plasmids - genetics Production of active biomolecules Proteins Recombinant Proteins - metabolism sak gene SAK protein Serine Endopeptidases - genetics Serine Endopeptidases - metabolism Staphylococcus aureus staphylokinase Transformation, Bacterial wprA gene |
| title | Enhancement of secretion and extracellular stability of staphylokinase in Bacillus subtilis by wprA gene disruption |
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