Artificial covalent linkage of bacterial acyl carrier proteins for fatty acid production
Abstract
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.
- Authors:
- Publication Date:
- Research Org.:
- Brookhaven National Laboratory (BNL), Upton, NY (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES)
- OSTI Identifier:
- 1619617
- Alternate Identifier(s):
- OSTI ID: 1576227
- Report Number(s):
- BNL-212366-2019-JAAM
Journal ID: ISSN 2045-2322; 16011; PII: 52344
- Grant/Contract Number:
- SC0012704
- Resource Type:
- Published Article
- Journal Name:
- Scientific Reports
- Additional Journal Information:
- Journal Name: Scientific Reports Journal Volume: 9 Journal Issue: 1; Journal ID: ISSN 2045-2322
- Publisher:
- Nature Publishing Group
- Country of Publication:
- United Kingdom
- Language:
- English
- Subject:
- 60 APPLIED LIFE SCIENCES
Citation Formats
Rullán-Lind, Carlos, Ortiz-Rosario, Melissa, García-González, Andrea, Stojanoff, Vivian, Chorna, Nataliya E., Pietri, Ruth B., and Baerga-Ortiz, Abel. Artificial covalent linkage of bacterial acyl carrier proteins for fatty acid production. United Kingdom: N. p., 2019.
Web. doi:10.1038/s41598-019-52344-w.
Rullán-Lind, Carlos, Ortiz-Rosario, Melissa, García-González, Andrea, Stojanoff, Vivian, Chorna, Nataliya E., Pietri, Ruth B., & Baerga-Ortiz, Abel. Artificial covalent linkage of bacterial acyl carrier proteins for fatty acid production. United Kingdom. https://doi.org/10.1038/s41598-019-52344-w
Rullán-Lind, Carlos, Ortiz-Rosario, Melissa, García-González, Andrea, Stojanoff, Vivian, Chorna, Nataliya E., Pietri, Ruth B., and Baerga-Ortiz, Abel. Tue .
"Artificial covalent linkage of bacterial acyl carrier proteins for fatty acid production". United Kingdom. https://doi.org/10.1038/s41598-019-52344-w.
@article{osti_1619617,
title = {Artificial covalent linkage of bacterial acyl carrier proteins for fatty acid production},
author = {Rullán-Lind, Carlos and Ortiz-Rosario, Melissa and García-González, Andrea and Stojanoff, Vivian and Chorna, Nataliya E. and Pietri, Ruth B. and Baerga-Ortiz, Abel},
abstractNote = {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.},
doi = {10.1038/s41598-019-52344-w},
journal = {Scientific Reports},
number = 1,
volume = 9,
place = {United Kingdom},
year = {Tue Nov 05 00:00:00 EST 2019},
month = {Tue Nov 05 00:00:00 EST 2019}
}
Free Publicly Available Full Text
Publisher's Version of Record
https://doi.org/10.1038/s41598-019-52344-w
https://doi.org/10.1038/s41598-019-52344-w
Other availability
Search WorldCat to find libraries that may hold this journal
Cited by: 1 work
Citation information provided by
Web of Science
Web of Science
Save to My Library
You must Sign In or Create an Account in order to save documents to your library.
Works referenced in this record:
Biochemical characterization of polyunsaturated fatty acid synthesis in Schizochytrium: Release of the products as free fatty acids
journal, June 2009
- Metz, James G.; Kuner, Jerry; Rosenzweig, Bradley
- Plant Physiology and Biochemistry, Vol. 47, Issue 6
Loading of malonyl-CoA onto tandem acyl carrier protein domains of polyunsaturated fatty acid synthases
journal, June 2018
- Santín, Omar; Moncalián, Gabriel
- Journal of Biological Chemistry, Vol. 293, Issue 32
DAMMIF, a program for rapid ab-initio shape determination in small-angle scattering
journal, January 2009
- Franke, Daniel; Svergun, Dmitri I.
- Journal of Applied Crystallography, Vol. 42, Issue 2, p. 342-346
Small-Angle X-Ray Scattering on Biological Macromolecules and Nanocomposites in Solution
journal, April 2013
- Blanchet, Clement E.; Svergun, Dmitri I.
- Annual Review of Physical Chemistry, Vol. 64, Issue 1
Enhanced production of polyunsaturated fatty acids by enzyme engineering of tandem acyl carrier proteins
journal, October 2016
- Hayashi, Shohei; Satoh, Yasuharu; Ujihara, Tetsuro
- Scientific Reports, Vol. 6, Issue 1
Structure and regulation of the omega-3 polyunsaturated fatty acid synthase genes from the deep-sea bacterium Photobacterium profundum strain SS9
journal, June 2002
- Allen, Eric E.; Bartlett, Douglas H.
- Microbiology, Vol. 148, Issue 6
Determination of the regularization parameter in indirect-transform methods using perceptual criteria
journal, August 1992
- Svergun, D. I.
- Journal of Applied Crystallography, Vol. 25, Issue 4, p. 495-503
PRIMUS: a Windows PC-based system for small-angle scattering data analysis
journal, September 2003
- Konarev, Petr V.; Volkov, Vladimir V.; Sokolova, Anna V.
- Journal of Applied Crystallography, Vol. 36, Issue 5, p. 1277-1282
Identification of Claisen cyclase domain in fungal polyketide synthase WA, a naphthopyrone synthase of Aspergillus nidulans
journal, February 2001
- Fujii, Isao; Watanabe, Akira; Sankawa, Ushio
- Chemistry & Biology, Vol. 8, Issue 2
Production of Polyunsaturated Fatty Acids by Polyketide Synthases in Both Prokaryotes and Eukaryotes
journal, July 2001
- Metz, J. G.
- Science, Vol. 293, Issue 5528
Tandem Acyl Carrier Proteins in the Curacin Biosynthetic Pathway Promote Consecutive Multienzyme Reactions with a Synergistic Effect
journal, February 2011
- Gu, Liangcai; Eisman, Eli B.; Dutta, Somnath
- Angewandte Chemie International Edition, Vol. 50, Issue 12
A practical guide to small angle X-ray scattering (SAXS) of flexible and intrinsically disordered proteins
journal, August 2015
- Kikhney, Alexey G.; Svergun, Dmitri I.
- FEBS Letters, Vol. 589, Issue 19PartA
A Rapid Method of Total Lipid Extraction and Purification
journal, January 1959
- Bligh, E. G.; Dyer, W. J.
- Canadian Journal of Biochemistry and Physiology, Vol. 37, Issue 1
The I-TASSER Suite: protein structure and function prediction
journal, December 2014
- Yang, Jianyi; Yan, Renxiang; Roy, Ambrish
- Nature Methods, Vol. 12, Issue 1
Solution Structure of the Tandem Acyl Carrier Protein Domains from a Polyunsaturated Fatty Acid Synthase Reveals Beads-on-a-String Configuration
journal, February 2013
- Trujillo, Uldaeliz; Vázquez-Rosa, Edwin; Oyola-Robles, Delise
- PLoS ONE, Vol. 8, Issue 2
The Role of Tandem Acyl Carrier Protein Domains in Polyunsaturated Fatty Acid Biosynthesis
journal, May 2008
- Jiang, Hui; Zirkle, Ross; Metz, James G.
- Journal of the American Chemical Society, Vol. 130, Issue 20
Polyketide biosynthesis: a millennium review
journal, January 2001
- Staunton, James; Weissman, Kira J.
- Natural Product Reports, Vol. 18, Issue 4
The Unmodified (Apo) Form of Escherichia coli Acyl Carrier Protein Is a Potent Inhibitor of Cell Growth
journal, September 1995
- Keating, David H.; Carey, Merriann Rawlings; Cronan, John E.
- Journal of Biological Chemistry, Vol. 270, Issue 38