High-level expression of soluble subunit b of $ F_{1} $$ F_{0} $ ATP synthase in Escherichia coli cell-free system
Abstract The overexpression of subunit b of $ F_{1} $$ F_{0} $ adenosine triphosphate (ATP) synthase from Escherichia coli is so toxic that it even prevents the transformation of plasmids encoding this protein into E. coli BL21 (DE3). In the present work, E. coli cell-free system was chosen as an al...
Ausführliche Beschreibung
Autor*in: |
Lian, Jiazhang [verfasserIn] |
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Format: |
Artikel |
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Sprache: |
Englisch |
Erschienen: |
2009 |
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Anmerkung: |
© Springer-Verlag 2009 |
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Übergeordnetes Werk: |
Enthalten in: Applied microbiology and biotechnology - Springer Berlin Heidelberg, 1984, 85(2009), 2 vom: 11. Juni, Seite 303-311 |
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Übergeordnetes Werk: |
volume:85 ; year:2009 ; number:2 ; day:11 ; month:06 ; pages:303-311 |
Links: |
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DOI / URN: |
10.1007/s00253-009-2055-z |
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Katalog-ID: |
OLC2050724993 |
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10.1007/s00253-009-2055-z doi (DE-627)OLC2050724993 (DE-He213)s00253-009-2055-z-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Lian, Jiazhang verfasserin aut High-level expression of soluble subunit b of $ F_{1} $$ F_{0} $ ATP synthase in Escherichia coli cell-free system 2009 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag 2009 Abstract The overexpression of subunit b of $ F_{1} $$ F_{0} $ adenosine triphosphate (ATP) synthase from Escherichia coli is so toxic that it even prevents the transformation of plasmids encoding this protein into E. coli BL21 (DE3). In the present work, E. coli cell-free system was chosen as an alternative to express this highly toxic membrane protein. This protein was either produced as precipitates followed by detergent resolubilization or expressed as a soluble form with detergent addition. Among several types of tested detergents, Brij 58 could effectively solubilize approximately 85% of the target membrane protein within a wide range of concentration (48 to 178 times critical micelle concentration [CMC]) with little effect on the expression level. With the presence of Brij 58 at the final concentration of 96 times CMC in the E. coli cell-free system, 789 μg/mL of soluble subunit b was achieved after 4 h biosynthesis, which is the highest level for the expression of membrane proteins in a batch-mode cell-free expression system. The present work provides a rapid and efficient procedure of expressing one membrane protein with high cytotoxicity in the cell-free system and will be helpful to further exploration of reconstituting $ F_{1} $$ F_{0} $ ATP synthase into liposome or polymer vesicle to design a nanoelectromechanical system device. Subunit of ATP synthase ATP synthase Cell-free expression system Membrane protein Ma, Yi aut Cai, Jin aut Wu, Ming aut Wang, Jufang aut Wang, Xiaoning aut Xu, Zhinan aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 85(2009), 2 vom: 11. Juni, Seite 303-311 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:85 year:2009 number:2 day:11 month:06 pages:303-311 https://doi.org/10.1007/s00253-009-2055-z lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_21 GBV_ILN_23 GBV_ILN_40 GBV_ILN_69 GBV_ILN_70 GBV_ILN_100 GBV_ILN_130 GBV_ILN_147 GBV_ILN_267 GBV_ILN_285 GBV_ILN_2004 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 GBV_ILN_4307 AR 85 2009 2 11 06 303-311 |
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10.1007/s00253-009-2055-z doi (DE-627)OLC2050724993 (DE-He213)s00253-009-2055-z-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Lian, Jiazhang verfasserin aut High-level expression of soluble subunit b of $ F_{1} $$ F_{0} $ ATP synthase in Escherichia coli cell-free system 2009 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag 2009 Abstract The overexpression of subunit b of $ F_{1} $$ F_{0} $ adenosine triphosphate (ATP) synthase from Escherichia coli is so toxic that it even prevents the transformation of plasmids encoding this protein into E. coli BL21 (DE3). In the present work, E. coli cell-free system was chosen as an alternative to express this highly toxic membrane protein. This protein was either produced as precipitates followed by detergent resolubilization or expressed as a soluble form with detergent addition. Among several types of tested detergents, Brij 58 could effectively solubilize approximately 85% of the target membrane protein within a wide range of concentration (48 to 178 times critical micelle concentration [CMC]) with little effect on the expression level. With the presence of Brij 58 at the final concentration of 96 times CMC in the E. coli cell-free system, 789 μg/mL of soluble subunit b was achieved after 4 h biosynthesis, which is the highest level for the expression of membrane proteins in a batch-mode cell-free expression system. The present work provides a rapid and efficient procedure of expressing one membrane protein with high cytotoxicity in the cell-free system and will be helpful to further exploration of reconstituting $ F_{1} $$ F_{0} $ ATP synthase into liposome or polymer vesicle to design a nanoelectromechanical system device. Subunit of ATP synthase ATP synthase Cell-free expression system Membrane protein Ma, Yi aut Cai, Jin aut Wu, Ming aut Wang, Jufang aut Wang, Xiaoning aut Xu, Zhinan aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 85(2009), 2 vom: 11. Juni, Seite 303-311 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:85 year:2009 number:2 day:11 month:06 pages:303-311 https://doi.org/10.1007/s00253-009-2055-z lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_21 GBV_ILN_23 GBV_ILN_40 GBV_ILN_69 GBV_ILN_70 GBV_ILN_100 GBV_ILN_130 GBV_ILN_147 GBV_ILN_267 GBV_ILN_285 GBV_ILN_2004 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 GBV_ILN_4307 AR 85 2009 2 11 06 303-311 |
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10.1007/s00253-009-2055-z doi (DE-627)OLC2050724993 (DE-He213)s00253-009-2055-z-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Lian, Jiazhang verfasserin aut High-level expression of soluble subunit b of $ F_{1} $$ F_{0} $ ATP synthase in Escherichia coli cell-free system 2009 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag 2009 Abstract The overexpression of subunit b of $ F_{1} $$ F_{0} $ adenosine triphosphate (ATP) synthase from Escherichia coli is so toxic that it even prevents the transformation of plasmids encoding this protein into E. coli BL21 (DE3). In the present work, E. coli cell-free system was chosen as an alternative to express this highly toxic membrane protein. This protein was either produced as precipitates followed by detergent resolubilization or expressed as a soluble form with detergent addition. Among several types of tested detergents, Brij 58 could effectively solubilize approximately 85% of the target membrane protein within a wide range of concentration (48 to 178 times critical micelle concentration [CMC]) with little effect on the expression level. With the presence of Brij 58 at the final concentration of 96 times CMC in the E. coli cell-free system, 789 μg/mL of soluble subunit b was achieved after 4 h biosynthesis, which is the highest level for the expression of membrane proteins in a batch-mode cell-free expression system. The present work provides a rapid and efficient procedure of expressing one membrane protein with high cytotoxicity in the cell-free system and will be helpful to further exploration of reconstituting $ F_{1} $$ F_{0} $ ATP synthase into liposome or polymer vesicle to design a nanoelectromechanical system device. Subunit of ATP synthase ATP synthase Cell-free expression system Membrane protein Ma, Yi aut Cai, Jin aut Wu, Ming aut Wang, Jufang aut Wang, Xiaoning aut Xu, Zhinan aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 85(2009), 2 vom: 11. Juni, Seite 303-311 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:85 year:2009 number:2 day:11 month:06 pages:303-311 https://doi.org/10.1007/s00253-009-2055-z lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_21 GBV_ILN_23 GBV_ILN_40 GBV_ILN_69 GBV_ILN_70 GBV_ILN_100 GBV_ILN_130 GBV_ILN_147 GBV_ILN_267 GBV_ILN_285 GBV_ILN_2004 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 GBV_ILN_4307 AR 85 2009 2 11 06 303-311 |
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10.1007/s00253-009-2055-z doi (DE-627)OLC2050724993 (DE-He213)s00253-009-2055-z-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Lian, Jiazhang verfasserin aut High-level expression of soluble subunit b of $ F_{1} $$ F_{0} $ ATP synthase in Escherichia coli cell-free system 2009 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag 2009 Abstract The overexpression of subunit b of $ F_{1} $$ F_{0} $ adenosine triphosphate (ATP) synthase from Escherichia coli is so toxic that it even prevents the transformation of plasmids encoding this protein into E. coli BL21 (DE3). In the present work, E. coli cell-free system was chosen as an alternative to express this highly toxic membrane protein. This protein was either produced as precipitates followed by detergent resolubilization or expressed as a soluble form with detergent addition. Among several types of tested detergents, Brij 58 could effectively solubilize approximately 85% of the target membrane protein within a wide range of concentration (48 to 178 times critical micelle concentration [CMC]) with little effect on the expression level. With the presence of Brij 58 at the final concentration of 96 times CMC in the E. coli cell-free system, 789 μg/mL of soluble subunit b was achieved after 4 h biosynthesis, which is the highest level for the expression of membrane proteins in a batch-mode cell-free expression system. The present work provides a rapid and efficient procedure of expressing one membrane protein with high cytotoxicity in the cell-free system and will be helpful to further exploration of reconstituting $ F_{1} $$ F_{0} $ ATP synthase into liposome or polymer vesicle to design a nanoelectromechanical system device. Subunit of ATP synthase ATP synthase Cell-free expression system Membrane protein Ma, Yi aut Cai, Jin aut Wu, Ming aut Wang, Jufang aut Wang, Xiaoning aut Xu, Zhinan aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 85(2009), 2 vom: 11. Juni, Seite 303-311 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:85 year:2009 number:2 day:11 month:06 pages:303-311 https://doi.org/10.1007/s00253-009-2055-z lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_21 GBV_ILN_23 GBV_ILN_40 GBV_ILN_69 GBV_ILN_70 GBV_ILN_100 GBV_ILN_130 GBV_ILN_147 GBV_ILN_267 GBV_ILN_285 GBV_ILN_2004 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 GBV_ILN_4307 AR 85 2009 2 11 06 303-311 |
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10.1007/s00253-009-2055-z doi (DE-627)OLC2050724993 (DE-He213)s00253-009-2055-z-p DE-627 ger DE-627 rakwb eng 570 VZ 12 ssgn BIODIV DE-30 fid Lian, Jiazhang verfasserin aut High-level expression of soluble subunit b of $ F_{1} $$ F_{0} $ ATP synthase in Escherichia coli cell-free system 2009 Text txt rdacontent ohne Hilfsmittel zu benutzen n rdamedia Band nc rdacarrier © Springer-Verlag 2009 Abstract The overexpression of subunit b of $ F_{1} $$ F_{0} $ adenosine triphosphate (ATP) synthase from Escherichia coli is so toxic that it even prevents the transformation of plasmids encoding this protein into E. coli BL21 (DE3). In the present work, E. coli cell-free system was chosen as an alternative to express this highly toxic membrane protein. This protein was either produced as precipitates followed by detergent resolubilization or expressed as a soluble form with detergent addition. Among several types of tested detergents, Brij 58 could effectively solubilize approximately 85% of the target membrane protein within a wide range of concentration (48 to 178 times critical micelle concentration [CMC]) with little effect on the expression level. With the presence of Brij 58 at the final concentration of 96 times CMC in the E. coli cell-free system, 789 μg/mL of soluble subunit b was achieved after 4 h biosynthesis, which is the highest level for the expression of membrane proteins in a batch-mode cell-free expression system. The present work provides a rapid and efficient procedure of expressing one membrane protein with high cytotoxicity in the cell-free system and will be helpful to further exploration of reconstituting $ F_{1} $$ F_{0} $ ATP synthase into liposome or polymer vesicle to design a nanoelectromechanical system device. Subunit of ATP synthase ATP synthase Cell-free expression system Membrane protein Ma, Yi aut Cai, Jin aut Wu, Ming aut Wang, Jufang aut Wang, Xiaoning aut Xu, Zhinan aut Enthalten in Applied microbiology and biotechnology Springer Berlin Heidelberg, 1984 85(2009), 2 vom: 11. Juni, Seite 303-311 (DE-627)129942634 (DE-600)392453-1 (DE-576)015507750 0175-7598 nnns volume:85 year:2009 number:2 day:11 month:06 pages:303-311 https://doi.org/10.1007/s00253-009-2055-z lizenzpflichtig Volltext GBV_USEFLAG_A SYSFLAG_A GBV_OLC FID-BIODIV SSG-OLC-TEC SSG-OLC-CHE SSG-OLC-PHA SSG-OLC-DE-84 GBV_ILN_21 GBV_ILN_23 GBV_ILN_40 GBV_ILN_69 GBV_ILN_70 GBV_ILN_100 GBV_ILN_130 GBV_ILN_147 GBV_ILN_267 GBV_ILN_285 GBV_ILN_2004 GBV_ILN_2018 GBV_ILN_4012 GBV_ILN_4082 GBV_ILN_4277 GBV_ILN_4305 GBV_ILN_4307 AR 85 2009 2 11 06 303-311 |
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Enthalten in Applied microbiology and biotechnology 85(2009), 2 vom: 11. Juni, Seite 303-311 volume:85 year:2009 number:2 day:11 month:06 pages:303-311 |
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Enthalten in Applied microbiology and biotechnology 85(2009), 2 vom: 11. Juni, Seite 303-311 volume:85 year:2009 number:2 day:11 month:06 pages:303-311 |
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High-level expression of soluble subunit b of $ F_{1} $$ F_{0} $ ATP synthase in Escherichia coli cell-free system |
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Lian, Jiazhang |
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high-level expression of soluble subunit b of $ f_{1} $$ f_{0} $ atp synthase in escherichia coli cell-free system |
title_auth |
High-level expression of soluble subunit b of $ F_{1} $$ F_{0} $ ATP synthase in Escherichia coli cell-free system |
abstract |
Abstract The overexpression of subunit b of $ F_{1} $$ F_{0} $ adenosine triphosphate (ATP) synthase from Escherichia coli is so toxic that it even prevents the transformation of plasmids encoding this protein into E. coli BL21 (DE3). In the present work, E. coli cell-free system was chosen as an alternative to express this highly toxic membrane protein. This protein was either produced as precipitates followed by detergent resolubilization or expressed as a soluble form with detergent addition. Among several types of tested detergents, Brij 58 could effectively solubilize approximately 85% of the target membrane protein within a wide range of concentration (48 to 178 times critical micelle concentration [CMC]) with little effect on the expression level. With the presence of Brij 58 at the final concentration of 96 times CMC in the E. coli cell-free system, 789 μg/mL of soluble subunit b was achieved after 4 h biosynthesis, which is the highest level for the expression of membrane proteins in a batch-mode cell-free expression system. The present work provides a rapid and efficient procedure of expressing one membrane protein with high cytotoxicity in the cell-free system and will be helpful to further exploration of reconstituting $ F_{1} $$ F_{0} $ ATP synthase into liposome or polymer vesicle to design a nanoelectromechanical system device. © Springer-Verlag 2009 |
abstractGer |
Abstract The overexpression of subunit b of $ F_{1} $$ F_{0} $ adenosine triphosphate (ATP) synthase from Escherichia coli is so toxic that it even prevents the transformation of plasmids encoding this protein into E. coli BL21 (DE3). In the present work, E. coli cell-free system was chosen as an alternative to express this highly toxic membrane protein. This protein was either produced as precipitates followed by detergent resolubilization or expressed as a soluble form with detergent addition. Among several types of tested detergents, Brij 58 could effectively solubilize approximately 85% of the target membrane protein within a wide range of concentration (48 to 178 times critical micelle concentration [CMC]) with little effect on the expression level. With the presence of Brij 58 at the final concentration of 96 times CMC in the E. coli cell-free system, 789 μg/mL of soluble subunit b was achieved after 4 h biosynthesis, which is the highest level for the expression of membrane proteins in a batch-mode cell-free expression system. The present work provides a rapid and efficient procedure of expressing one membrane protein with high cytotoxicity in the cell-free system and will be helpful to further exploration of reconstituting $ F_{1} $$ F_{0} $ ATP synthase into liposome or polymer vesicle to design a nanoelectromechanical system device. © Springer-Verlag 2009 |
abstract_unstemmed |
Abstract The overexpression of subunit b of $ F_{1} $$ F_{0} $ adenosine triphosphate (ATP) synthase from Escherichia coli is so toxic that it even prevents the transformation of plasmids encoding this protein into E. coli BL21 (DE3). In the present work, E. coli cell-free system was chosen as an alternative to express this highly toxic membrane protein. This protein was either produced as precipitates followed by detergent resolubilization or expressed as a soluble form with detergent addition. Among several types of tested detergents, Brij 58 could effectively solubilize approximately 85% of the target membrane protein within a wide range of concentration (48 to 178 times critical micelle concentration [CMC]) with little effect on the expression level. With the presence of Brij 58 at the final concentration of 96 times CMC in the E. coli cell-free system, 789 μg/mL of soluble subunit b was achieved after 4 h biosynthesis, which is the highest level for the expression of membrane proteins in a batch-mode cell-free expression system. The present work provides a rapid and efficient procedure of expressing one membrane protein with high cytotoxicity in the cell-free system and will be helpful to further exploration of reconstituting $ F_{1} $$ F_{0} $ ATP synthase into liposome or polymer vesicle to design a nanoelectromechanical system device. © Springer-Verlag 2009 |
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High-level expression of soluble subunit b of $ F_{1} $$ F_{0} $ ATP synthase in Escherichia coli cell-free system |
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