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Solubilization of the overexpressed integral membrane protein alkane monooxygenase of the recombinant Escherichia coli W3110[pGEc47]
Abstract
The integral membrane-bound alkane monooxygenase (AlkB) from Pseudomonas oleovorans has been overexpressed in the recombinant Escherichia coli strain W3110[pGEc47] and expression levels of 10 to 15% relative to the total cell protein were reached. The amount of phospholipids in induced cells is about 3-fold higher compared to the wild-type and AlkB has been shown to be located in small membrane vesicles. We present here a study on the solubilization of these AlkB containing membrane vesicles by different detergents with special emphasis on structural requirements for a surfactant preserving the activity of AlkB. Moreover, the effects of the detergents used on the complete alkane hydroxylase system was studied.
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Microorganisms use oxygenase reactions to create more biodegradable forms of substrates.
“Basically, micro-organisms utilize the oxygenase reactions to create more biodegradable forms of substrates.”
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Araujo Granda, P., Gras, A., & Ginovart, M. (2016) · Computational and Structural Biotechnology Journal · doi:10.1016/j.csbj.2016.08.001 · Cited by 9
Metadata
- Authors
- Peters, J., & Witholt, B.
- Affiliations
- ETH Zurich (CH)
- Corresponding author
- Witholt, Bernard
- Venue
- Biochimica et Biophysica Acta (BBA) - Biomembranes
- Publisher
- Elsevier BV
- Published
- 1994-12
- Volume
- 1196
- Issue
- 2
- Pages
- 145-153
- ISSN
- 0005-2736 (ISSN-L)
- OpenAlex
- W2041760905
- Cited by
- 9
- References
- 0
- Citations by year
- 2018: 1 · 2016: 1
- FWCI
- 0.95
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochimica et Biophysica Acta (BBA) - Biomembranes (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Physical Sciences › Environmental Science › Pollution › Microbial bioremediation and biosurfactants
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biotechnology › Enzyme Production and Characterization
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Enzyme Catalysis and Immobilization
- Keywords
- alkane monooxygenase, AlkB, Pseudomonas oleovorans, membrane vesicles, detergent solubilization, surfactant activity, phospholipids, recombinant protein overexpression, Escherichia coli W3110
- MeSH terms
- Bacterial Outer Membrane Proteins, Cytochrome P-450 Enzyme System, Detergents, Escherichia coli, Mixed Function Oxygenases, Pseudomonas, Recombination, Genetic, Solubility, Cytochrome P-450 CYP4A
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