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Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein
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| Zeitschriftentitel: | Applied and Environmental Microbiology |
|---|---|
| Personen und Körperschaften: | , , |
| In: | Applied and Environmental Microbiology, 73, 2007, 19, S. 6036-6044 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
American Society for Microbiology
|
| Schlagwörter: |
| author_facet |
Malone, Cheryl L. Boles, Blaise R. Horswill, Alexander R. Malone, Cheryl L. Boles, Blaise R. Horswill, Alexander R. |
|---|---|
| author |
Malone, Cheryl L. Boles, Blaise R. Horswill, Alexander R. |
| spellingShingle |
Malone, Cheryl L. Boles, Blaise R. Horswill, Alexander R. Applied and Environmental Microbiology Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein Ecology Applied Microbiology and Biotechnology Food Science Biotechnology |
| author_sort |
malone, cheryl l. |
| spelling |
Malone, Cheryl L. Boles, Blaise R. Horswill, Alexander R. 0099-2240 1098-5336 American Society for Microbiology Ecology Applied Microbiology and Biotechnology Food Science Biotechnology http://dx.doi.org/10.1128/aem.00912-07 <jats:title>ABSTRACT</jats:title> <jats:p> The Agr quorum-sensing system of <jats:italic>Staphylococcus aureus</jats:italic> modulates the expression of virulence factors in response to autoinducing peptides (AIPs). The peptides are seven to nine residues in length and have the C-terminal five residues constrained in a thiolactone ring. We have developed a new method to generate AIP structures using an engineered DnaB mini-intein from <jats:italic>Synechocystis</jats:italic> sp. strain PCC6803. In the method, an oligonucleotide encoding the AIP is ligated to the intein and the fusion protein is expressed and purified by affinity chromatography. To produce the correct AIP structure, intein splicing is interrupted, allowing the cysteine side chain to catalyze thiolactone ring formation and release AIP from the resin. The technique is simple and robust, and we have successfully produced the three main classes of AIPs using the intein system. The intein-generated AIPs possessed the correct thiolactone ring modification based on biochemical analysis, and, importantly, all the samples were bioactive against <jats:italic>S. aureus</jats:italic> . The AIP activity was confirmed through Agr interference and activation profiling with developed <jats:italic>S. aureus</jats:italic> reporter strains. The simplicity of the method, benefits of DNA encoding, and scalable nature enable the production of <jats:italic>S. aureus</jats:italic> AIPs for many biological applications. </jats:p> Biosynthesis of <i>Staphylococcus aureus</i> Autoinducing Peptides by Using the <i>Synechocystis</i> DnaB Mini-Intein Applied and Environmental Microbiology |
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10.1128/aem.00912-07 |
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American Society for Microbiology, 2007 |
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American Society for Microbiology, 2007 |
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| title |
Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein |
| title_unstemmed |
Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein |
| title_full |
Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein |
| title_fullStr |
Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein |
| title_full_unstemmed |
Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein |
| title_short |
Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein |
| title_sort |
biosynthesis of
<i>staphylococcus aureus</i>
autoinducing peptides by using the
<i>synechocystis</i>
dnab mini-intein |
| topic |
Ecology Applied Microbiology and Biotechnology Food Science Biotechnology |
| url |
http://dx.doi.org/10.1128/aem.00912-07 |
| publishDate |
2007 |
| physical |
6036-6044 |
| description |
<jats:title>ABSTRACT</jats:title>
<jats:p>
The Agr quorum-sensing system of
<jats:italic>Staphylococcus aureus</jats:italic>
modulates the expression of virulence factors in response to autoinducing peptides (AIPs). The peptides are seven to nine residues in length and have the C-terminal five residues constrained in a thiolactone ring. We have developed a new method to generate AIP structures using an engineered DnaB mini-intein from
<jats:italic>Synechocystis</jats:italic>
sp. strain PCC6803. In the method, an oligonucleotide encoding the AIP is ligated to the intein and the fusion protein is expressed and purified by affinity chromatography. To produce the correct AIP structure, intein splicing is interrupted, allowing the cysteine side chain to catalyze thiolactone ring formation and release AIP from the resin. The technique is simple and robust, and we have successfully produced the three main classes of AIPs using the intein system. The intein-generated AIPs possessed the correct thiolactone ring modification based on biochemical analysis, and, importantly, all the samples were bioactive against
<jats:italic>S. aureus</jats:italic>
. The AIP activity was confirmed through Agr interference and activation profiling with developed
<jats:italic>S. aureus</jats:italic>
reporter strains. The simplicity of the method, benefits of DNA encoding, and scalable nature enable the production of
<jats:italic>S. aureus</jats:italic>
AIPs for many biological applications.
</jats:p> |
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| author | Malone, Cheryl L., Boles, Blaise R., Horswill, Alexander R. |
| author_facet | Malone, Cheryl L., Boles, Blaise R., Horswill, Alexander R., Malone, Cheryl L., Boles, Blaise R., Horswill, Alexander R. |
| author_sort | malone, cheryl l. |
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| description | <jats:title>ABSTRACT</jats:title> <jats:p> The Agr quorum-sensing system of <jats:italic>Staphylococcus aureus</jats:italic> modulates the expression of virulence factors in response to autoinducing peptides (AIPs). The peptides are seven to nine residues in length and have the C-terminal five residues constrained in a thiolactone ring. We have developed a new method to generate AIP structures using an engineered DnaB mini-intein from <jats:italic>Synechocystis</jats:italic> sp. strain PCC6803. In the method, an oligonucleotide encoding the AIP is ligated to the intein and the fusion protein is expressed and purified by affinity chromatography. To produce the correct AIP structure, intein splicing is interrupted, allowing the cysteine side chain to catalyze thiolactone ring formation and release AIP from the resin. The technique is simple and robust, and we have successfully produced the three main classes of AIPs using the intein system. The intein-generated AIPs possessed the correct thiolactone ring modification based on biochemical analysis, and, importantly, all the samples were bioactive against <jats:italic>S. aureus</jats:italic> . The AIP activity was confirmed through Agr interference and activation profiling with developed <jats:italic>S. aureus</jats:italic> reporter strains. The simplicity of the method, benefits of DNA encoding, and scalable nature enable the production of <jats:italic>S. aureus</jats:italic> AIPs for many biological applications. </jats:p> |
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| imprint | American Society for Microbiology, 2007 |
| imprint_str_mv | American Society for Microbiology, 2007 |
| institution | DE-L229, DE-D275, DE-Bn3, DE-Brt1, DE-D161, DE-Zwi2, DE-Gla1, DE-Zi4, DE-15, DE-Pl11, DE-Rs1, DE-105, DE-14, DE-Ch1 |
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| language | English |
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| physical | 6036-6044 |
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| source_id | 49 |
| spelling | Malone, Cheryl L. Boles, Blaise R. Horswill, Alexander R. 0099-2240 1098-5336 American Society for Microbiology Ecology Applied Microbiology and Biotechnology Food Science Biotechnology http://dx.doi.org/10.1128/aem.00912-07 <jats:title>ABSTRACT</jats:title> <jats:p> The Agr quorum-sensing system of <jats:italic>Staphylococcus aureus</jats:italic> modulates the expression of virulence factors in response to autoinducing peptides (AIPs). The peptides are seven to nine residues in length and have the C-terminal five residues constrained in a thiolactone ring. We have developed a new method to generate AIP structures using an engineered DnaB mini-intein from <jats:italic>Synechocystis</jats:italic> sp. strain PCC6803. In the method, an oligonucleotide encoding the AIP is ligated to the intein and the fusion protein is expressed and purified by affinity chromatography. To produce the correct AIP structure, intein splicing is interrupted, allowing the cysteine side chain to catalyze thiolactone ring formation and release AIP from the resin. The technique is simple and robust, and we have successfully produced the three main classes of AIPs using the intein system. The intein-generated AIPs possessed the correct thiolactone ring modification based on biochemical analysis, and, importantly, all the samples were bioactive against <jats:italic>S. aureus</jats:italic> . The AIP activity was confirmed through Agr interference and activation profiling with developed <jats:italic>S. aureus</jats:italic> reporter strains. The simplicity of the method, benefits of DNA encoding, and scalable nature enable the production of <jats:italic>S. aureus</jats:italic> AIPs for many biological applications. </jats:p> Biosynthesis of <i>Staphylococcus aureus</i> Autoinducing Peptides by Using the <i>Synechocystis</i> DnaB Mini-Intein Applied and Environmental Microbiology |
| spellingShingle | Malone, Cheryl L., Boles, Blaise R., Horswill, Alexander R., Applied and Environmental Microbiology, Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein, Ecology, Applied Microbiology and Biotechnology, Food Science, Biotechnology |
| title | Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein |
| title_full | Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein |
| title_fullStr | Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein |
| title_full_unstemmed | Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein |
| title_short | Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein |
| title_sort | biosynthesis of <i>staphylococcus aureus</i> autoinducing peptides by using the <i>synechocystis</i> dnab mini-intein |
| title_unstemmed | Biosynthesis of Staphylococcus aureus Autoinducing Peptides by Using the Synechocystis DnaB Mini-Intein |
| topic | Ecology, Applied Microbiology and Biotechnology, Food Science, Biotechnology |
| url | http://dx.doi.org/10.1128/aem.00912-07 |