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Efficient Production of Active Polyhydroxyalkanoate Synthase in Escherichia coli by Coexpression of Molecular Chaperones.

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Title: Efficient Production of Active Polyhydroxyalkanoate Synthase in Escherichia coli by Coexpression of Molecular Chaperones.
Authors: Thomson, Nicholas M.1, Saika, Azusa2, Ushimaru, Kazunori2, Sangiambut, Smith1, Tsuge, Takeharu2, Summers, David K.3, Sivaniah, Easan1 es10009@cam.ac.uk
Source: Applied & Environmental Microbiology. Mar2013, Vol. 79 Issue 6, p1948-1955. 8p.
Subjects: Polyhydroxyalkanoates, Necator, Escherichia coli, Molecular chaperones, Enzyme activation
Abstract: The type I polyhydroxyalkanoate synthase from Cupriavidus necator was heterologously expressed in Escherichia coli with simultaneous overexpression of chaperone proteins. Compared to expression of synthase alone (14.55 mg liter-1), coexpression with chaperones resulted in the production of larger total quantities of enzyme, including a larger proportion in the soluble fraction. The largest increase was seen when the GroEL/GroES system was coexpressed, resulting in approximately 6-fold-greater enzyme yields (82.37 mg liter-1) than in the absence of coexpressed chaperones. The specific activity of the purified enzyme was unaffected by coexpression with chaperones. Therefore, the increase in yield was attributed to an enhanced soluble fraction of synthase. Chaperones were also coexpressed with a polyhydroxyalkanoate production operon, resulting in the production of polymers with generally reduced molecular weights. This suggests a potential use for chaperones to control the physical properties of the polymer. [ABSTRACT FROM AUTHOR]
Copyright of Applied & Environmental Microbiology is the property of American Society for Microbiology and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
Database: Engineering Source
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DbLabel: Engineering Source
An: 86667554
AccessLevel: 6
PubType: Academic Journal
PubTypeId: academicJournal
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  Data: Efficient Production of Active Polyhydroxyalkanoate Synthase in Escherichia coli by Coexpression of Molecular Chaperones.
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  Data: <searchLink fieldCode="AR" term="%22Thomson%2C+Nicholas+M%2E%22">Thomson, Nicholas M.</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Saika%2C+Azusa%22">Saika, Azusa</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Ushimaru%2C+Kazunori%22">Ushimaru, Kazunori</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Sangiambut%2C+Smith%22">Sangiambut, Smith</searchLink><relatesTo>1</relatesTo><br /><searchLink fieldCode="AR" term="%22Tsuge%2C+Takeharu%22">Tsuge, Takeharu</searchLink><relatesTo>2</relatesTo><br /><searchLink fieldCode="AR" term="%22Summers%2C+David+K%2E%22">Summers, David K.</searchLink><relatesTo>3</relatesTo><br /><searchLink fieldCode="AR" term="%22Sivaniah%2C+Easan%22">Sivaniah, Easan</searchLink><relatesTo>1</relatesTo><i> es10009@cam.ac.uk</i>
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  Data: <searchLink fieldCode="JN" term="%22Applied+%26+Environmental+Microbiology%22">Applied & Environmental Microbiology</searchLink>. Mar2013, Vol. 79 Issue 6, p1948-1955. 8p.
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  Data: <searchLink fieldCode="DE" term="%22Polyhydroxyalkanoates%22">Polyhydroxyalkanoates</searchLink><br /><searchLink fieldCode="DE" term="%22Necator%22">Necator</searchLink><br /><searchLink fieldCode="DE" term="%22Escherichia+coli%22">Escherichia coli</searchLink><br /><searchLink fieldCode="DE" term="%22Molecular+chaperones%22">Molecular chaperones</searchLink><br /><searchLink fieldCode="DE" term="%22Enzyme+activation%22">Enzyme activation</searchLink>
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  Data: The type I polyhydroxyalkanoate synthase from Cupriavidus necator was heterologously expressed in Escherichia coli with simultaneous overexpression of chaperone proteins. Compared to expression of synthase alone (14.55 mg liter-1), coexpression with chaperones resulted in the production of larger total quantities of enzyme, including a larger proportion in the soluble fraction. The largest increase was seen when the GroEL/GroES system was coexpressed, resulting in approximately 6-fold-greater enzyme yields (82.37 mg liter-1) than in the absence of coexpressed chaperones. The specific activity of the purified enzyme was unaffected by coexpression with chaperones. Therefore, the increase in yield was attributed to an enhanced soluble fraction of synthase. Chaperones were also coexpressed with a polyhydroxyalkanoate production operon, resulting in the production of polymers with generally reduced molecular weights. This suggests a potential use for chaperones to control the physical properties of the polymer. [ABSTRACT FROM AUTHOR]
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  Data: <i>Copyright of Applied & Environmental Microbiology is the property of American Society for Microbiology and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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        Value: 10.1128/AEM.02881-12
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      – Code: eng
        Text: English
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        PageCount: 8
        StartPage: 1948
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      – SubjectFull: Polyhydroxyalkanoates
        Type: general
      – SubjectFull: Necator
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      – SubjectFull: Escherichia coli
        Type: general
      – SubjectFull: Molecular chaperones
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      – SubjectFull: Enzyme activation
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      – TitleFull: Efficient Production of Active Polyhydroxyalkanoate Synthase in Escherichia coli by Coexpression of Molecular Chaperones.
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              Text: Mar2013
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              Y: 2013
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