Initiation of fatty acid biosynthesis in Pseudomonas putida KT2440

DOI: 10.1016/j.ymben.2023.02.006

Authors: Kevin J. Mcnaught (National Renewable Energy Laboratory (USA); DOE Agile BioFoundry) , Eugene Kuatsjah (National Renewable Energy Laboratory (USA); Oak Ridge National Laboratory) , Michael Zahn (University of Portsmouth) , Érica T. Prates (Oak Ridge National Laboratory) , Huiling Shao (University of California Los Angeles) , Gayle J. Bentley (National Renewable Energy Laboratory (USA); DOE Agile BioFoundry) , Andrew R. Pickford (University of Portsmouth) , Josephine N. Gruber (National Renewable Energy Laboratory (USA)) , Kelley V. Hestmark (National Renewable Energy Laboratory (USA); DOE Agile BioFoundry) , Daniel A. Jacobson (Oak Ridge National Laboratory) , Brenton C. Poirier (DOE Agile BioFoundry; Pacific Northwest National Laboratory) , Chen Ling (National Renewable Energy Laboratory (USA); DOE Agile BioFoundry) , Myrsini San Marchi (University of California Los Angeles) , William E. Michener (National Renewable Energy Laboratory (USA)) , Carrie D. Nicora (Pacific Northwest National Laboratory) , Jacob N. Sanders (University of California Los Angeles) , Caralyn J. Szostkiewicz (National Renewable Energy Laboratory (USA); DOE Agile BioFoundry) , Dušan Veličković (Pacific Northwest National Laboratory) , Mowei Zhou (Pacific Northwest National Laboratory) , Nathalie Munoz (DOE Agile BioFoundry; Pacific Northwest National Laboratory) , Young-Mo Kim (DOE Agile BioFoundry; Pacific Northwest National Laboratory) , Jon K. Magnuson (DOE Agile BioFoundry; Pacific Northwest National Laboratory) , Kristin E. Burnum-Johnson (DOE Agile BioFoundry; Pacific Northwest National Laboratory) , Kendall N. Houk (University of California Los Angeles) , John E. Mcgeehan (University of Portsmouth) , Christopher W. Johnson (National Renewable Energy Laboratory (USA); DOE Agile BioFoundry) , Gregg T. Beckham (National Renewable Energy Laboratory (USA); DOE Agile BioFoundry; Oak Ridge National Laboratory)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Metabolic Engineering , VOL 3

State: Published (Approved)
Published: February 2023
Diamond Proposal Number(s): 23269

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 modelling 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.

Journal Keywords: Fatty acid biosynthesis; Decarboxylase; Hotdog fold; Pseudomonas putida

Diamond Keywords: Bacteria; Enzymes

Subject Areas: Biology and Bio-materials, Medicine


Instruments: I03-Macromolecular Crystallography

Added On: 20/02/2023 09:13

Discipline Tags:

Pathogens Biotechnology Health & Wellbeing Structural biology Engineering & Technology Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)