Microbial communities play a vital role in biogeochemical cycles, allowing the biodegradation of a wide range of pollutants. The composition of the community and the interactions between its members affect degradation rate and determine the identity of the final products. Here, we demonstrate the application of sequencing technologies and metabolic modeling approaches towards enhancing biodegradation of atrazine-a herbicide causing environmental pollution. Treatment of agriculture soil with atrazine is shown to induce significant changes in community structure and functional performances. Genome-scale metabolic models were constructed for Arthrobacter, the atrazine degrader, and four other non-atrazine degrading species whose relative abundance in soil was changed following exposure to the herbicide. By modeling community function we show that consortia including the direct degrader and non-degrader differentially abundant species perform better than Arthrobacter alone. Simulations predict that growth/degradation enhancement is derived by metabolic exchanges between community members. Based on simulations we designed endogenous consortia optimized for enhanced degradation whose performances were validated in vitro and biostimulation strategies that were tested in pot experiments. Overall, our analysis demonstrates that understanding community function in its wider context, beyond the single direct degrader perspective, promotes the design of biostimulation strategies.These authors contributed equally:
Atrazine is an herbicide and a pollutant of great environmental concern that is naturally biodegraded by microbial communities. Paenarthrobacter aurescens TC1 is one of the most studied degraders of this herbicide. Here, we developed a genome scale metabolic model for P. aurescens TC1, iRZ1179, to study the atrazine degradation process at organism level. Constraint based flux balance analysis and time dependent simulations were used to explore the organism's phenotypic landscape. Simulations aimed at designing media optimized for supporting growth and enhancing degradation, by passing the need in strain design via genetic modifications. Growth and degradation simulations were carried with more than 100 compounds consumed by P. aurescens TC1. In vitro validation confirmed the predicted classification of different compounds as efficient, moderate or poor stimulators of growth. Simulations successfully captured previous reports on the use of glucose and phosphate as biostimulators of atrazine degradation, supported by in vitro validation. Model predictions can go beyond supplementing the medium with a single compound and can predict the growth outcomes for higher complexity combinations. Hence, the analysis demonstrates that the exhaustive power of the genome scale metabolic reconstruction allows capturing complexities that are beyond common biochemical expertise and knowledge and further support the importance of computational platforms for the educated design of complex media. The model presented here can potentially serve as a predictive tool towards achieving optimal biodegradation efficiencies and for the development of ecologically friendly solutions for pollutant degradation. Atrazine (2-chloro-4-ethylamino-6-isopropylamino-1,3,5-triazine) is an herbicide employed to control broadleaf and grass mainly in crops such as rice, wheat, maize, and sorghum. It is also a well-known pollutant of great environmental concern. Atrazine has been shown to have negative effects such as DNA damage, gene expression shifts, cancer and endocrine disruption 1,2. Its residues are found in soil samples decades after it was last applied and were shown to chronically leach into local aquifers 3. As such, efforts are being made to limit and monitor its use 4. While atrazine was banned in the European Union and Switzerland since 2003, the United States Environmental Protection Agency still allows its wide use under monitoring 5,6. Areas contaminated with atrazine and other hazardous herbicides are rapidly increasing worldwide introducing a need in remediation approaches. Bioremediation-an environmental bioprocess in which naturally occurring organisms are used for breaking down hazardous substances into less toxic or non-toxic substances-is increasingly acknowledged as a cost-effective feasible alternative for environmental cleaning 7-9. Critical environmental bioprocesses are naturally carried out by bacteria and are related to removal of pollutants from water, soil or air 10. Thus, bacterial bioremediation is widely applied for the ...
Atrazine is an herbicide and pollutant of great environmental concern that is naturally biodegraded by microbial communities. The efficiency of biodegradation can be improved through the stimulating addition of fertilizers, electron acceptors, etc. In recent years, metabolic modelling approaches have become widely used as an in silico tool for organism-level phenotyping and the subsequent development of metabolic engineering strategies including biodegradation improvement. Here, we constructed a genome scale metabolic model, iRZ960, for Paenarthrobacter aurescens TC1 – a widely studied atrazine degrader - aiming at simulating its degradation activity. A mathematical stoichiometric metabolic model was constructed based on a published genome sequence of P. aurescens TC1. An Initial draft model was automatically constructed using the RAST and KBase servers. The draft was developed into a predictive model through semi-automatic gap-filling procedures including manual curation. In addition to growth predictions under different conditions, model simulations were used to identify optimized media for enhancing the natural degradation of atrazine without a need in strain design via genetic modifications. Model predictions for growth and atrazine degradation efficiency were tested in myriad of media supplemented with different combinations of carbon and nitrogen sources that were verified in vitro. Experimental validations support the reliability of the model’s predictions for both bacterial growth (biomass accumulation) and atrazine degradation. Predictive tools, such as the presented model, can be applied for achieving optimal biodegradation efficiencies and for the development of ecologically friendly solutions for pollutant degradation in changing environments.
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