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Instruments / Technical Area	Publication Details	Published	Actions
I03-Macromolecular Crystallography	Initiation of fatty acid biosynthesis in Pseudomonas putida KT2440 Kevin J. Mcnaught , Eugene Kuatsjah , Michael Zahn , Érica T. Prates , Huiling Shao , Gayle J. Bentley , Andrew R. Pickford , Josephine N. Gruber , Kelley V. Hestmark , Daniel A. Jacobson , Brenton C. Poirier , Chen Ling , Myrsini San Marchi , William E. Michener , Carrie D. Nicora , Jacob N. Sanders , Caralyn J. Szostkiewicz , Dušan Veličković , Mowei Zhou , Nathalie Munoz , Young-Mo Kim , Jon K. Magnuson , Kristin E. Burnum-Johnson , Kendall N. Houk , John E. Mcgeehan , Christopher W. Johnson , Gregg T. Beckham Metabolic Engineering 3 DOI: 10.1016/j.ymben.2023.02.006 Diamond Proposal Number(s): [23269] Open Access Show Abstract ▼ 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.	Feb 2023

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I03-Macromolecular Crystallography

Initiation of fatty acid biosynthesis in Pseudomonas putida KT2440