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Potential of ectomycorrhizal fungus Pisolithus tinctorius to tolerate and to degrade trifluoroacetate into fluoroform
New Biotechnology · Volume 31S · July 2014
SYMPOSIUM 21: BIODEGRADATION AND BIOMEDIATION
O21-4
O21-5
Potential of ectomycorrhizal fungus Pisolithus tinctorius to tolerate and to degrade trifluoroacetate into fluoroform
Evaluation of the biological sulfamethoxazole degradation mechanism for biotechnological applications
Paula Castro los Afonso 3
1,∗
2
2
3
, Albina Franco , Miguel Ramos , Sara Cravo , Car-
1
CBQF – Centro de Biotecnologia e Química Fina – Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa/Porto, Portugal 2 CBQF – Centro de Biotecnologia e Química Fina – Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Portugal 3 2CEQUIMED-UP, Laboratório de Química Orgânica e Farmacêutica, Centro Interdisciplinar de Investigac¸ão Marinha e Ambiental (CIIMAR/CIMAR), Departamento Ciências Químicas, Faculdade de Farmácia, Universidade do Porto, Portugal
Trifluoroacetate (TFA) is a persistent fluorinated organic compound originated from the degradation of fluorinated compounds, such as HCFC and isoflurane, or as a side product from the thermolysis of fluoropolymers, like Teflon. TFA can reach soil through precipitation, where it persists in water and soil, and may contribute to forest decline. In this study, we assessed the capacity of P. tinctorius, an ectomycorrhizal fungus (ECMF), to tolerate and/or degrade TFA. In vitro studies in glucose-supplemented solid medium showed that the fungus tolerated up to 8.77 mM TFA. P. tinctorius also degraded 88.3%, 89.9%, and 42.1% of 0.88, 2.39, and 4.39 mM TFA, respectively, in liquid cultures. No TFA accumulation was detected on the fungus mycelium, suggesting that TFA depletion was due to fungal degradation. Defluorination was not detected. A volatile compound with a structure and behavior compatible with fluoroform (CHF3 ), a potent greenhouse gas, was detected using GC-MS/MS, only in the gas phase of sealed P. tinctorius cultures supplied with TFA. Further confirmation of this compound is needed. Nevertheless, the study shows that P. tinctorius was capable to degrade TFA possibly through a similar pathway to that found on marine sediments. The results evidence the role of ECMF may play in the degradation of fluorinated organic compounds, enhancing their potential contribution on establishing tree growth in soils exposed to organic contamination. Acknowledgements: A. Franco thanks FCT the grant SFRH/BD/47722/2008. This work was supported by FCT Project -PTDC-AGR-CFL-111583-2009 and PEst-OE/EQB/LA0016/2011. http://dx.doi.org/10.1016/j.nbt.2014.05.1760
Benjamin Ricken 1,∗ , Markus Lenz 1 , Danuta Cichocka 1 , Hans-Peter Kohler 2 , Boris Kolvenbach 1 , Philippe Francois-Xavier Corvini 1 1
Institute for Ecopreneurship/University of Applied Sciences and Art Northwestern Switzerland, Switzerland 2 Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Microbiology, Switzerland
Sulfonamide antibiotics are of rising concern as their release into the environment has been suspected to enhance the formation of resistant pathogenic bacterial strains [1]. Among sulfonamides, antibiotics such as sulfamethoxazole (SMX) are arousing public interest since they are photo- and thermostable as well as recalcitrant to traditional, biological waste water treatment processes [2]. Our studies focused on SMX as it is the most used sulfonamide antibiotic in human medicine. Even though extensive research has been done on SMX degradation by bacterial consortia or isolates, the complete pathway is not yet understood, let alone the proteins involved. We were able to identify the first crucial degradation step of several sulfonamide antibiotics [3] by means of isolating Microbacterium sp. strain BR1, proven to partially mineralize SMX for the first time [4]. Concerning the pathway, molecular oxygen is incorporated into the sulfonamide antibiotic at its aniline moiety through ipso-substitution, which is an uncommon process for biodegradation. This leads to the release of the verified metabolites: 3-amino-5-methylisoxazole, p-aminophenol and sulfite. Further investigations are currently under way in order to identify the involved protein and its cofactor dependencies. To conclude, it is postulated that there is one common sulfonamide degradation mechanism among various bacteria [3]. Its elucidation and the determination of involved proteins are essential for upcoming biological sulfonamide removal approaches. References [1].Hruska, Franek. Vet Med 2012;2012:1–35. [2].Gros, et al. Environ Int 2010;36:15–26. [3].Ricken, et al. Appl Environ Microbiol 2013;79:5550–8. [4].Bouju, et al. Appl Environ Microbiol 2012;78:277–9.
http://dx.doi.org/10.1016/j.nbt.2014.05.1761
www.elsevier.com/locate/nbt S65
SYMPOSIUM 21: BIODEGRADATION AND BIOMEDIATION
O21-4
O21-5
Potential of ectomycorrhizal fungus Pisolithus tinctorius to tolerate and to degrade trifluoroacetate into fluoroform
Evaluation of the biological sulfamethoxazole degradation mechanism for biotechnological applications
Paula Castro los Afonso 3
1,∗
2
2
3
, Albina Franco , Miguel Ramos , Sara Cravo , Car-
1
CBQF – Centro de Biotecnologia e Química Fina – Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa/Porto, Portugal 2 CBQF – Centro de Biotecnologia e Química Fina – Laboratório Associado, Escola Superior de Biotecnologia, Universidade Católica Portuguesa, Portugal 3 2CEQUIMED-UP, Laboratório de Química Orgânica e Farmacêutica, Centro Interdisciplinar de Investigac¸ão Marinha e Ambiental (CIIMAR/CIMAR), Departamento Ciências Químicas, Faculdade de Farmácia, Universidade do Porto, Portugal
Trifluoroacetate (TFA) is a persistent fluorinated organic compound originated from the degradation of fluorinated compounds, such as HCFC and isoflurane, or as a side product from the thermolysis of fluoropolymers, like Teflon. TFA can reach soil through precipitation, where it persists in water and soil, and may contribute to forest decline. In this study, we assessed the capacity of P. tinctorius, an ectomycorrhizal fungus (ECMF), to tolerate and/or degrade TFA. In vitro studies in glucose-supplemented solid medium showed that the fungus tolerated up to 8.77 mM TFA. P. tinctorius also degraded 88.3%, 89.9%, and 42.1% of 0.88, 2.39, and 4.39 mM TFA, respectively, in liquid cultures. No TFA accumulation was detected on the fungus mycelium, suggesting that TFA depletion was due to fungal degradation. Defluorination was not detected. A volatile compound with a structure and behavior compatible with fluoroform (CHF3 ), a potent greenhouse gas, was detected using GC-MS/MS, only in the gas phase of sealed P. tinctorius cultures supplied with TFA. Further confirmation of this compound is needed. Nevertheless, the study shows that P. tinctorius was capable to degrade TFA possibly through a similar pathway to that found on marine sediments. The results evidence the role of ECMF may play in the degradation of fluorinated organic compounds, enhancing their potential contribution on establishing tree growth in soils exposed to organic contamination. Acknowledgements: A. Franco thanks FCT the grant SFRH/BD/47722/2008. This work was supported by FCT Project -PTDC-AGR-CFL-111583-2009 and PEst-OE/EQB/LA0016/2011. http://dx.doi.org/10.1016/j.nbt.2014.05.1760
Benjamin Ricken 1,∗ , Markus Lenz 1 , Danuta Cichocka 1 , Hans-Peter Kohler 2 , Boris Kolvenbach 1 , Philippe Francois-Xavier Corvini 1 1
Institute for Ecopreneurship/University of Applied Sciences and Art Northwestern Switzerland, Switzerland 2 Eawag, Swiss Federal Institute of Aquatic Science and Technology, Department of Environmental Microbiology, Switzerland
Sulfonamide antibiotics are of rising concern as their release into the environment has been suspected to enhance the formation of resistant pathogenic bacterial strains [1]. Among sulfonamides, antibiotics such as sulfamethoxazole (SMX) are arousing public interest since they are photo- and thermostable as well as recalcitrant to traditional, biological waste water treatment processes [2]. Our studies focused on SMX as it is the most used sulfonamide antibiotic in human medicine. Even though extensive research has been done on SMX degradation by bacterial consortia or isolates, the complete pathway is not yet understood, let alone the proteins involved. We were able to identify the first crucial degradation step of several sulfonamide antibiotics [3] by means of isolating Microbacterium sp. strain BR1, proven to partially mineralize SMX for the first time [4]. Concerning the pathway, molecular oxygen is incorporated into the sulfonamide antibiotic at its aniline moiety through ipso-substitution, which is an uncommon process for biodegradation. This leads to the release of the verified metabolites: 3-amino-5-methylisoxazole, p-aminophenol and sulfite. Further investigations are currently under way in order to identify the involved protein and its cofactor dependencies. To conclude, it is postulated that there is one common sulfonamide degradation mechanism among various bacteria [3]. Its elucidation and the determination of involved proteins are essential for upcoming biological sulfonamide removal approaches. References [1].Hruska, Franek. Vet Med 2012;2012:1–35. [2].Gros, et al. Environ Int 2010;36:15–26. [3].Ricken, et al. Appl Environ Microbiol 2013;79:5550–8. [4].Bouju, et al. Appl Environ Microbiol 2012;78:277–9.
http://dx.doi.org/10.1016/j.nbt.2014.05.1761
www.elsevier.com/locate/nbt S65