Initiation of Fatty Acid Biosynthesis in Pseudomonas putida KT2440

Kevin McNaught, Eugene Kuatsjah, Michael Zahn, Erica Prates, Huiling Shao, Gayle Bentley, Andrew Pickford, Josephine Gruber, Kelley Hestmark, Daniel Jacobson, Brenton Poirier, Chen Ling, Myrsini San Marchi, William Michener, Carrie Nicora, Jacob Sanders, Caralyn Szostkiewicz, Dusan Velickovic, Mowei Zhou, Nathalie MunozYoung-Mo Kim, Jon Magnuson, Kristin Burnum-Johnson, K. Houk, John McGeehan, Christopher Johnson, Gregg Beckham

Research output: Contribution to journalArticlepeer-review

8 Scopus Citations

Abstract

Deciphering the mechanisms of bacterial fatty acid biosynthesis is crucial for both the engineering of bacterial hosts to produce fatty acid-derived molecules and the development of new antibiotics. However, gaps in our understanding of the initiation of fatty acid biosynthesis remain. Here, we demonstrate that the industrially relevant microbe Pseudomonas putida KT2440 contains three distinct pathways to initiate fatty acid biosynthesis. The first two routes employ conventional β-ketoacyl-ACP synthase III enzymes, FabH1 and FabH2, that accept short- and medium-chain-length acyl-CoAs, respectively. The third route utilizes a malonyl-ACP decarboxylase enzyme, MadB. A combination of exhaustive in vivo alanine-scanning mutagenesis, in vitro biochemical characterization, X-ray crystallography, and computational modeling elucidate the presumptive mechanism of malonyl-ACP decarboxylation via MadB. Given that functional homologs of MadB are widespread throughout domain Bacteria, this ubiquitous alternative fatty acid initiation pathway provides new opportunities to target a range of biotechnology and biomedical applications.

Original languageAmerican English
Pages (from-to)193-203
Number of pages11
JournalMetabolic Engineering
Volume76
DOIs
StatePublished - 2023

Bibliographical note

Publisher Copyright:
© 2023 The Authors

NREL Publication Number

  • NREL/JA-2A00-84894

Keywords

  • Decarboxylase
  • Fatty acid biosynthesis
  • Hotdog fold
  • Pseudomonas putida

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