8AYV

Crystal structure of the Malonyl-ACP Decarboxylase MadB from Pseudomonas putida

Summary for 8AYV

• Entry DOI: 10.2210/pdb8ayv/pdb
• Descriptor: YiiD_C domain-containing protein (2 entities in total)
• Functional Keywords: madb, fatty acid biosynthesis, malonyl-coa, transferase
• Biological source: Pseudomonas putida KT2440
• Total number of polymer chains: 2
• Total formula weight: 34072.64

Authors

Zahn, M.,Kuatsjah, E.,Beckham, G.T.,McGeehan, J.E. (deposition date: 2022-09-03, release date: 2023-03-01, Last modification date: 2024-05-01)

Primary citation

McNaught, K.J.,Kuatsjah, E.,Zahn, M.,Prates, E.T.,Shao, H.,Bentley, G.J.,Pickford, A.R.,Gruber, J.N.,Hestmark, K.V.,Jacobson, D.A.,Poirier, B.C.,Ling, C.,San Marchi, M.,Michener, W.E.,Nicora, C.D.,Sanders, J.N.,Szostkiewicz, C.J.,Velickovic, D.,Zhou, M.,Munoz, N.,Kim, Y.M.,Magnuson, J.K.,Burnum-Johnson, K.E.,Houk, K.N.,McGeehan, J.E.,Johnson, C.W.,Beckham, G.T.
Initiation of fatty acid biosynthesis in Pseudomonas putida KT2440.
Metab Eng, 76:193-203, 2023

PubMed 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.

PubMed: 36796578
DOI: 10.1016/j.ymben.2023.02.006

Experimental method

X-RAY DIFFRACTION (1.044 Å)