title: Ongoing functional evolution of the bacterial atrazine chlorohydrolase AtzA.
abstract: Triazine herbicides such as atrazine and simazine which were heavily used in the latter half of the twentieth century constituted a rich new source of nitrogen for soil microbes. An atzA dechlorinase active against both atrazine and simazine was isolated from various soil bacteria from diverse locations in the mid 1990s. We have surveyed the atzA genes from eight triazine-degrading Aminobacter aminovorans strains isolated from French agricultural soils recurrently exposed to triazines in 2000. Six amino acid differences from the original isolate were each found in more than one of the A. aminovorans strains. Three of these in particular (V92L, A170T and A296T) were recovered from a majority of the isolates and from locations separated by up to 900 km, so may reflect ongoing selection for the new function. Two of the latter (A170T and A296T) were indeed found to confer higher specificity for simazine, albeit not atrazine, and greater affinity for a metal ion required for activity, than did the original variant. In contrast, we found that ongoing maintenance of the original atzA-containing isolate in laboratory culture for 12 years in a medium containing high concentrations of atrazine has led to the fixation of another amino acid substitution that substantially reduces activity for the triazines. The high concentrations of atrazine in the medium may have relaxed the selection for a highly efficient triazine dechlorinase activity, and that there is some, as yet uncharacterised, counter selection against the activity of this enzyme under these conditions.
authors.affiliation: CSIRO Ecosystem Sciences, GPO Box 1700, Canberra, ACT, 2601, Australia.
authors.forename: Sajid
authors.initials: S
authors.lastname: Noor
authors.forename: Frédérique
authors.initials: F
authors.lastname: Changey
authors.forename: John G
authors.initials: JG
authors.lastname: Oakeshott
authors.forename: Colin
authors.initials: C
authors.lastname: Scott
authors.forename: Fabrice
authors.initials: F
authors.lastname: Martin-Laurent
mesh_terms: D019943:Amino Acid Substitution; D001280:Atrazine / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D001673:Biodegradation, Environmental; D003470:Culture Media; D019143:Evolution, Molecular; D006540:Herbicides / Q000378:metabolism*; D006867:Hydrolases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D008958:Models, Molecular; D009154:Mutation; D011549:Pseudomonas / Q000201:enzymology / Q000235:genetics* / Q000302:isolation & purification; D012839:Simazine / Q000378:metabolism*; D012988:Soil Microbiology; D013329:Structure-Activity Relationship; D013379:Substrate Specificity
chemical_list: D001426:Bacterial Proteins; D003470:Culture Media; D006540:Herbicides; D006867:Hydrolases; C100894:atrazine chlorohydrolase; D001280:Atrazine; D012839:Simazine