Genome-Wide Expression Analysis Unravels Fluoroalkane Metabolism in <i>Pseudomonas</i> sp. Strain 273
Yongchao Xie, Diana Ramirez, Gao Chen, Guang He, Yanchen Sun, Fadime Kara Murdoch, Frank E. Löffler
Abstract
Pseudomonas sp. strain 273 grows with medium-chain terminally fluorinated alkanes under oxic conditions, releases fluoride, and synthesizes long-chain fluorofatty acids. To shed light on the genes involved in fluoroalkane metabolism, genome, and transcriptome sequencing of strain 273 grown with 1,10-difluorodecane (DFD), decane, and acetate were performed. Strain 273 harbors three genes encoding putative alkane monooxygenases (AlkB), key enzymes for initiating alkane degradation. Transcripts of alkB -2 were significantly more abundant in both decane- and DFD-grown cells compared to acetate-grown cells, suggesting AlkB-2 catalyzes the attack on terminal CH 3 and CH 2 F groups. Coordinately expressed with alkB -2 was an adjacent gene encoding a fused ferredoxin–ferredoxin reductase (Fd–Fdr). Phylogenetic analysis distinguished AlkB that couples with fused Fd–Fdr reductases from AlkB with alternate architectures. A gene cluster containing an ( S )-2-haloacid dehalogenase ( had ) gene was up-regulated in cells grown with DFD, suggesting a possible role in the removal of the ω-fluorine. Genes involved in long-chain fatty acid biosynthesis were not differentially expressed during growth with acetate, decane, or DFD, suggesting the bacterium’s biosynthetic machinery does not discriminate against monofluoro-fatty acid intermediates. The analysis sheds first light on genes and catalysts involved in the microbial metabolism of fluoroalkanes.