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Artificial covalent linkage of bacterial acyl carrier proteins for fatty acid production.
- Source :
-
Scientific reports [Sci Rep] 2019 Nov 05; Vol. 9 (1), pp. 16011. Date of Electronic Publication: 2019 Nov 05. - Publication Year :
- 2019
-
Abstract
- Acyl carrier proteins (ACPs) are essential to the production of fatty acids. In some species of marine bacteria, ACPs are arranged into tandem repeats joined by peptide linkers, an arrangement that results in high fatty acid yields. By contrast, Escherichia coli, a relatively low producer of fatty acids, uses a single-domain ACP. In this work, we have engineered the native E. coli ACP into tandem di- and tri-domain constructs joined by a naturally occurring peptide linker from the PUFA synthase of Photobacterium profundum. The size of these tandem fused ACPs was determined by size exclusion chromatography to be higher (21 kDa, 36 kDa and 141 kDa) than expected based on the amino acid sequence (12 kDa, 24 kDa and 37 kDa, respectively) suggesting the formation of a flexible extended conformation. Structural studies using small-angle X-ray scattering (SAXS), confirmed this conformational flexibility. The thermal stability for the di- and tri-domain constructs was similar to that of the unfused ACP, indicating a lack of interaction between domains. Lastly, E. coli cultures harboring tandem ACPs produced up to 1.6 times more fatty acids than wild-type ACP, demonstrating the viability of ACP fusion as a method to enhance fatty acid yield in bacteria.
- Subjects :
- Acyl Carrier Protein chemistry
Acyl Carrier Protein genetics
Amino Acid Sequence
Bacterial Proteins chemistry
Bacterial Proteins genetics
Escherichia coli metabolism
Fatty Acids analysis
Gas Chromatography-Mass Spectrometry
Protein Conformation
Protein Stability
Recombinant Proteins biosynthesis
Recombinant Proteins chemistry
Recombinant Proteins isolation & purification
Scattering, Small Angle
Temperature
X-Ray Diffraction
Acyl Carrier Protein metabolism
Bacterial Proteins metabolism
Fatty Acids metabolism
Photobacterium metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 9
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 31690733
- Full Text :
- https://doi.org/10.1038/s41598-019-52344-w