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- Biological treatment of a contaminated gaseous emission from a leather industry in a suspended-growth bioreactorPublication . Carvalho, M. F.; Duque, A. F.; Moura, S. C.; Amorim, Catarina L.; Ferreira Jorge, R. M.; Castro, Paula M. L.A suspended-growth bioreactor (SGB) was operated for the treatment of a gaseous stream mimicking emissions generated at a leather industrial company. The main volatile organic compounds (VOCs) present in the gaseous stream consisted of 1-methoxy-2-propanol, 2,6-dimethyl-4-heptanone, 2-butoxyethanol, toluene and butylacetate. A microbial consortium able to degrade these VOCs was successfully enriched. A laboratory- scale SGB was established and operated for 210-d with an 8 h cycle period and with shutdowns at weekends. Along this period, the SGB was exposed to organic loads (OL) between 6.5 and 2.3 £ 102 g h¡1 m¡3. Most of the compounds were not detected at the outlet of the SGB. The highest total VOC removal efficiency (RE) (ca 99%) was observed when an OL of 1.6 £ 102 g h¡1 m¡3 was fed to the SGB. The maximum total VOC elimination capacity (1.8 £ 102 g h¡1 m¡3) was achieved when the OL applied to the SGB was 2.3 £ 102 g h¡1 m¡3. For all the operating conditions, the SGB showed high levels of degradation of toluene and butylacetate (RE t 100%). This study also revealed that recirculation of the gaseous effluent improved the performance of the SGB. Overall, the SGB was shown to be robust, showing high performance after night and weekend shutdown periods.
- Degradation of fluoroanilines by the wild strain Labrys portucalensisPublication . Amorim, Catarina L.; Carvalho, Maria F.; Afonso, Carlos M. M.; Castro, Paula M. L.Aromatic amine compounds, many of them with halogenated substituents, constitute a major class of environmental pollutants that have been released into soil and water due to extensive use in industries and agriculture. Biodegradation has been found to be a major route for the removal of this kind of toxic and recalcitrant pollutants from the environment. Whereas the degradation of mono- and dichlorinated anilines has been studied, very little is known about fluorinated anilines. Therefore, the objective of this study was to investigate, under aerobic conditions, the degradation of 2-, 3- and 4-fluoroaniline by a previously isolated pure bacterium, designated as strain F11. This microorganism, identified as Labrys portucalensis, was isolated from a contaminated site in northern Portugal and has the unique capacity to utilize fluorobenzene as a sole carbon and energy source,. The results of the biodegradation of 2-, 3- and 4-fluoroaniline by strain F11 showed that this microorganism is able to completely degrade 2- fluoroaniline and partially degrade 4-fluoroaniline, when these compounds are present as a sole carbon and energy source. Biodegradation of these two compounds also occurred, although at a lower rate, in the absence of an external nitrogen source in the culture medium. To our knowledge, this is the first study reporting the biodegradation of 2- and 4-fluoroaniline as a sole carbon and energy source by a pure microbial culture. C.L. Amorim and M.F. Carvalho wish to acknowledge a research grant from Fundação para a Ciência e Tecnologia (FCT), Portugal (Ref. SFRH/BD/47109/2008 and SFRH/BPD/44670/2008, respectively) and Fundo Social Europeu (III Quadro Comunitário de Apoio). This work was supported by the FCT Project - PTDC/BIO/67306/2006
- Mineralization of 4-fluorocinnamic acid by a Rhodococcus strainPublication . Amorim, Catarina L.; Ferreira, António César Silva; Carvalho, M.F.; Afonso, Carlos M. M.; Castro, Paula M.L.A bacterial strain capable of aerobic degradation of 4-fluorocinnamic acid (4-FCA) as the sole source of carbon and energy was isolated from a biofilm reactor operating for the treatment of 2-fluorophenol. The organism, designated as strain S2, was identified by 16S rRNA gene analysis as a member of the genus Rhodococcus. Strain S2 was able to mineralize 4-FCA as sole carbon and energy source. In the presence of a conventional carbon source (sodium acetate [SA]), growth rate of strain S2 was enhanced from 0.04 to 0.14 h−1 when the culture medium was fed with 0.5 mMof 4- FCA, and the time for complete removal of 4-FCA decreased from 216 to 50 h.When grown in SA-supplemented medium, 4-FCA concentrations up to 1 mM did not affect the length of the lag phase, and for 4-FCA concentrations up to 3 mM, strain S2 was able to completely remove the target fluorinated compound. 4-Fluorobenzoate (4-FBA) was transiently formed in the culture medium, reaching concentrations up to 1.7 mM when the cultures were supplemented with 3.5mMof 4-FCA. Trans,trans-muconate was also transiently formed as a metabolic intermediate. Compounds with molecular mass compatible with 3-carboxymuconate and 3-oxoadipate were also detected in the culture medium. Strain S2 was able to mineralize a range of other haloorganic compounds, including 2- fluorophenol, to which the biofilm reactor had been exposed. To our knowledge, this is the first time that mineralization of 4-FCA as the sole carbon source by a single bacterial culture is reported.
- Production of 4-fluorocatechol from fluorobenzene by the wild srain Labrys portucalensisPublication . Moreira, I. S.; Carvalho, M. F.; Amorim, Catarina L.; Afonso, C M. M.; Janssen, D. B.; Castro, P. M. L.
- Co-metabolic degradation of chlorobenzene by the fluorobenzene degrading wild strain Labrys portucalensisPublication . Moreira, Irina S.; Amorim, Catarina L.; Carvalho, Maria F.; Castro, Paula M.L.Due to the widespread use of chlorobenzene (CB), environmental contamination with this compound is a major concern. A previously isolated bacterium named Labrys portucalensis (strain F11), that is able to use fluorobenzene (FB) as sole carbon and energy source, was tested for its capability to degrade CB. Strain F11 was able to partially degrade CB only when F11 cells were previously grown in FB. Biodegradation of 0.5 mM of CB was achieved at a rate of 7.95 0.39 mmol l 1 day 1 with concomitant stoichiometric release of 50% of the chloride, while degradation of 1 mM of this compound resulted in 85% degradation at a rate of 16.9 0.81 mmol l 1 day 1 and with a 15% chloride release on the basis of the amount of compound biodegraded. Total CB biodegradation and dechlorination was only achieved when FB was also supplied to F11 cultures, suggesting cometabolic transformation. Total degradation of 0.5 mM of CB and 0.5 mM of FB occurred simultaneously at degradation rates of 105 6.07 mmol l 1 day 1 and 126 16.2 mmol l 1 day 1 respectively, with stoichiometric halogen release. Growth yield was lower when both substrates were present, suggesting synergistic inhibition. To our knowledge, this is the first time that co-metabolic biodegradation of CB in the presence of the fluorinated analogue is reported.
- Effect of the metals iron, copper and silver on fluorobenzene biodegradation by Labrys portucalensisPublication . Moreira, Irina S.; Amorim, Catarina L.; Carvalho, Maria F.; Ferreira, António C.; Afonso, Carlos M.; Castro, Paula M. L.Organic and metallic pollutants are ubiquitous in the environment. Many metals are reported to be toxic to microorganisms and to inhibit biodegradation. The effect of the metals iron, copper and silver on the metabolism of Labrys portucalensis F11 and on fluorobenzene (FB) biodegradation was examined. The results indicate that the addition of 1 mM of Fe2+ to the culture medium has a positive effect on bacterial growth and has no impact in the biodegradation of 1 and 2 mM of FB. The presence of 1 mM of Cu2+ was found to strongly inhibit the growth of F11 cultures and to reduce the biodegradation of 1 and 2 mM of FB to ca. 50 %, with 80 % of stoichiometrically expected fluoride released. In the experiments with resting cells, the FB degraded (from 2 mM supplied) was reduced ca. 20 % whereas the fluoride released was reduced to 45 % of that stoichiometrically expected. Ag+ was the most potent inhibitor of FB degradation. In experiments with growing cells, the addition of 1 mM of Ag+ to the culture medium containing 1 and 2 mM of FB resulted in no fluoride release, whereas FB degradation was only one third of that observed in control cultures. In the experiments with resting cells, the addition of Ag+ resulted in 25 % reduction in substrate degradation and fluoride release was only 20 % of that stoichiometrically expected. The accumulation of catechol and 4-fluorocatechol in cultures supplemented with Cu2+ or Ag+ suggest inhibition of the key enzyme of FB metabolism-catechol 1,2-dioxygenase.
- Degradation of difluorobenzenes by the wild strain Labrys portucalensisPublication . Moreira, I. S.; Carvalho, M. F.; Amorim, Catarina L.; Janssen, D. B.; Castro, Paula M. L.
- Biodegradation of fluoroanilines by the wild strain Labrys portucalensisPublication . Amorim, Catarina L.; Carvalho, Maria F.; Afonso, Carlos M. M.; Castro, Paula M. L.Aniline and halogenated anilines are known as widespread environmental toxic pollutants released into soil and water. In contrast to aniline, which is rapidly metabolized via catechol, halosubstituted anilines are more resistant to microbial attack. A fluorobenzene-degrading bacterium, Labrys portucalensis strain F11, was tested under different culture conditions for the degradation potential towards 2-, 3- and 4-fluoroaniline (2-, 3- and 4-FA). Strain F11 was able to use FAs as a source of carbon and nitrogen however, supplementation with a nitrogen source improved substrate consumption and its dehalogenation extent. When F11 cells were previously grown on fluorobenzene (FB), higher biodegradation rates were achieved for all isomers. Complete 2-FA biodegradation with stoichiometric fluoride release was achieved when FB-induced cells were used. On the other hand, the degradation of 3- and 4-FA was characterized by incomplete defluorination of the target compounds suggesting accumulation of fluorinated intermediates. F11 cultures simultaneously supplied with FB and the fluorinated anilines showed a concomitant degradation of both substrates, suggesting co-metabolic biodegradation. To our knowledge, this is the first time that biodegradation of 2- and 3-FA as a sole carbon and nitrogen source and co-metabolic degradation of FA isomers in the presence of a structural analogous compound is reported.
- Degradation of difluorobenzenes by the wild strain Labrys portucalensisPublication . Moreira, I.S.; Carvalho, M. F.; Amorim, Catarina L.; Janssen, D. B.; Castro, Paula M. L.
- Studies on the biodegradation of fluoroanilines by a single microorganismPublication . Amorim, Catarina L.; Carvalho, Maria F.; Afonso, Carlos M. M.; Castro, Paula M. L.Aromatic amines are considered important environmental pollutants due to their toxic and recalcitrant properties. The widespread use of these compounds in the industry and agriculture has led to their release to soils and waters. Microbial degradation has been found to be the major mechanism to avoid the accumulation of these pollutants in the environment. Literature studies on the biodegradation of halogenated anilines have mostly focused on chlorinated anilines and very little is known about utilization of fluorinated derivatives. This study focuses on the biodegradation of 2-, 3- and 4-fluoroaniline by a microbial strain (strain F11) identified as Labrys portucalensis. This microorganism has the capacity to degrade aerobically fluorobenzene as sole carbon and energy source. Non-induced cells and cells induced in fluorobenzene were used to examine the capacity of the degrading strain to metabolize the target compounds. Co-metabolic studies with fluorobenzene were also conducted. The experiments were run in batch mode at 25ºC with constant shaking. Growth, depletion of the target compounds and fluoride release were monitored. Strain F11 was capable to completely degrade 2-fluoroaniline. 3- and 4-fluoroaniline were degraded nearly to completion by F11 cultures induced with fluorobenzene and by cultures fed simultaneously with fluorobenzene. The fluoride liberation for those two fluorinated anilines occurred at different extents. C.L. Amorim and M.F. Carvalho wish to acknowledge a research grant from Fundação para a Ciência e Tecnologia (FCT), Portugal (Ref. SFRH/BD/47109/2008 and SFRH/BPD/44670/2008, respectively) and Fundo Social Europeu (III Quadro Comunitário de Apoio). This work was supported by the FCT Project - PTDC/BIO/67306/2006