Microbial consortia that degrade 2,4-DNT by interspecies metabolism: isolation and characterisation

Abstract

Two consortia, isolated by selective enrichment from a soil sample of a nitroaromatic-contaminated site, degraded 2,4-DNT as their sole nitrogen source without accumulating one or more detectable intermediates. Though originating from the same sample, the optimised consortia had no common members, indicating that selective enrichment resulted in different end points. Consortium 1 and consortium 2 contained four and six bacterial species respectively, but both had two members that were able to collectively degrade 2,4-DNT. Variovorax paradoxus VM685 (consortium 1) and Pseudomonas sp. VM908 (consortium 2) initiate the catabolism of 2,4-DNT by an oxidation step, thereby releasing nitrite and forming 4-methyl-5-nitrocatechol (4M5NC). Both strains contained a gene similar to the dntAa gene encoding 2,4-DNT dioxygenase. They subsequently metabolised 4M5NC to 2-hydroxy-5-methylquinone (2H5MQ) and nitrite, indicative of DntB or 4M5NC monooxygenase activity. A second consortium member, Pseudomonas marginalis VM683 (consortium 1) and P. aeruginosa VM903, Sphingomonas sp. VM904, Stenotrophomonas maltophilia VM905 or P. viridiflava VM907 (consortium 2), was found to be indispensable for efficient growth of the consortia on 2,4-DNT and for efficient metabolisation of the intermediates 4M5NC and 2H5MQ. Knowledge about the interactions in this step of the degradation pathway is rather limited. In addition, both consortia can use 2,4-DNT as sole nitrogen and carbon source. A gene similar to the dntD gene of Burkholderia sp. strain DNT that catalyses ring fission was demonstrated by DNA hybridisation in the second member strains. To our knowledge, this is the first time that consortia are shown to be necessary for 2,4-DNT degradation.