Experimental observation of molecular-weight growth by the reactions of o-benzyne with benzyl radicals.

The chemistry of ortho-benzyne (o-C6H4) is of fundamental importance due to its role as an essential molecular building block in molecular-weight growth reactions. Here, we report on an experimental investigation of the reaction of o-C6H4 with benzyl (C7H7) radicals in a well-controlled flash pyrolysis experiment using a resistively heated SiC microtubular reactor at temperatures of 800-1600 K and pressures near 30  torr. To this end, the reactants o-C6H4 and C7H7 were pyrolytically generated from 1,2-diiodobenzene and benzyl bromide, respectively. Using molecular-beam time-of-flight mass spectrometry, we found that o-C6H4 associates with the benzyl to form C13H11 radicals, which decompose at higher temperatures via H-loss to form closed-shell C13H10 molecules. Our experimental results agree with earlier theoretical calculations by Matsugi and Miyoshi [Phys. Chem. Chem. Phys., 2012, 14, 9722-9728], who predicted the formation of fluorene (C13H10) + H to be the dominant reaction channel. At temperatures above 1400 K, we also observed the formation of C13H9 radicals, most likely the resonance-stabilized fluorenyl π-radical. Our study confirms that molecular-mass growth via the o-C6H4 + C7H7 reaction provides a versatile pathway for introducing five-membered rings, and hence curved structures, into polycyclic aromatic hydrocarbons.

Initiation of fatty acid biosynthesis in Pseudomonas putida KT2440.

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 ß-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.