Pseudomonas aeruginosa mutants affected in anaerobic growth on arginine: evidence for a four-gene cluster encoding the arginine deiminase pathway

Abstract

Pseudomonas aeruginosa PAO was able to grow in the absence of exogenous terminal electron acceptors, provided that the medium contained 30 to 40 mM L-arginine and 0.4% yeast extract. Under strictly anaerobic conditions (02 at <1 ppm), growth could be measured as an increase in protein and proceeded in a nonexponential way; arginine was largely converted to ornithine but not entirely consumed at the end of growth. In the GasPak anaerobic jar (Becton Dickinson and Co.), the wild-type strain PAOI grew on arginine-yeast extract medium in 3 to 5 days; mutants could be isolated that were unable to grow under these conditions. All mutants (except one) were defective in at least one of the three enzymes of the arginine deiminase pathway (arcA, arc), and arcC mutants) or in a novel function that might be involved in anaerobic arginine uptake (arcD mutants). The mutations arcA (arginine deiminase), arcB (catabolic ornithine carbamoyltransferase), arcC (carbantate kinase), and arcD were highly cotransducible and mapped in the 17-min chromosome region. Some mutations in the arc cluster led to low, noninducible levels of all three arginine deiminase pathway enzymes and thus may affect control elements required for induction of the postulated arc operon. Two fluorescent pseudomonads (P. putida and P. fluorescens) and P. mendocina, as well as one PAO mtitant, possessed an inducible arginine deiminase pathway and yet were unable to grow fermentatively on arginine. The ability to use arginine-derived ATP for growth may provide P. aeruginosa with a selective advantage when oxygen and nitrate are scarce.

Response of Pseudomonas aeruginosa PAO1 to low shear modelled microgravity involves AlgU regulation

“…Other than the activator of nitrate reduction, NarL, none of the gene transcripts involved in denitrification were more abundant in LSMMG than NG. Pseudomonas aeruginosa is able to ferment arginine and pyruvate as carbon and energy source in anoxic conditions (Vander Wauven et al, 1984;Schreiber et al, 2006). The arginine deiminase pathway, under control of Anr, is comprised of the genes arcABC (encoding enzymes), and arcD, encoding an arginine-ornithine antiporter (Luthi et al, 1986;Haas et al, 1987).…”

Diversity and anaerobic growth ofPseudomonasspp. isolated from modified atmosphere packaged minced beef

“…It has also been reported that P. aeruginosa is capable of different kinds of anaerobic fermentation (Arai 2011). In the absence of nitrate, P. aeruginosa ferments arginine via arginine deiminase pathway to conserve energy (Shoesmith and Sherris 1960;Vander Wauven et al 1984;Benkert et al 2008). Arginine is highly abundant in beef (Schweigert and Payne 1956;Holl o et al 2001a) hence arginine fermentation could be an explanation for the observed growth of Pseudomonas.…”

Catabolic ornithine carbamoyltransferase of Pseudomonas aeruginosa

“…1. The composition of growth media and selective antibiotic concentrations were as described previously (Vander Wauven et al, 1984;Jeenes et al, 1986;Luthi et al, 1986).…”

“…This enzyme is a trimer consisting of identical subunits of M,34000 (Itoh et al, 1988). The catabolic OTCase is the second enzyme of the arginine deiminase pathway, which is encoded by the arcDABC operon and allows ATP synthesis in the absence of respiration (Mercenier et al, 1980;Vander Wauven et al, 1984;Liithi et al, 1990). The catabolic OTCase has 12 identical subunits of Mr38000…”