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Title: Artificial covalent linkage of bacterial acyl carrier proteins for fatty acid production

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:
 [1];  [1];  [1];  [2];  [1];  [3];  [1]
  1. Univ. of Puerto Rico, San Juan (Puerto Rico)
  2. Brookhaven National Lab. (BNL), Upton, NY (United States)
  3. Univ. of Puerto Rico, Cayey (Puerto Rico)
Publication Date:
Research Org.:
Brookhaven National Lab. (BNL), Upton, NY (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1576227
Report Number(s):
BNL-212366-2019-JAAM
Journal ID: ISSN 2045-2322
Grant/Contract Number:  
SC0012704
Resource Type:
Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United States
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 States: 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 States. doi: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 States. doi:10.1038/s41598-019-52344-w. https://www.osti.gov/servlets/purl/1576227.
@article{osti_1576227,
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 = {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 States},
year = {2019},
month = {11}
}

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