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Aydin Berenjian - One of the best experts on this subject based on the ideXlab platform.
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effects of medium components in a glycerol based medium on vitamin k menaquinone 7 production by bacillus subtilis natto in biofilm reactors
Bioprocess and Biosystems Engineering, 2019Co-Authors: Ehsan Mahdinia, Ali Demirci, Aydin BerenjianAbstract:Menaquinone-7 (MK-7) as the most important form of Vitamin K has been reported to have miraculous benefits such as preventing cardiovascular diseases and osteoporosis along with antitumor effects. Therefore, there have been numerous studies in the past decades to improve MK-7 production via microbial fermentation. Unfortunately, both solid and liquid state fermentation strategies that are utilized for MK-7 production, face fundamental operational and scale-up issues as well as intense heat and mass transfer problems during fermentation. In this regard, biofilm reactors seem to be a practical solution to overcome these issues and enhance the production in agitated liquid fermentation. Therefore, this study was undertaken to utilize biofilm reactors in investigating and optimizing different media components in a glycerol-based medium. Using response surface methodology, the effects of glycerol, yeast extract, and soytone were studied in the fermentation medium on MK-7 production in biofilm reactor. With a composition of 48.2 g/L of glycerol, 8.1 g/L of yeast extracts, 13.6 g/L of soytone and 0.06 g/L of K2HPO4, MK-7 concentrations could reach 14.7 ± 1.4 mg/L in biofilm reactors, which was 57% higher compared to the MK-7 concentration achieved in Suspended-Cell reactors under similar conditions, while glycerol was depleted by the end of the fifth day in biofilm reactors, but glycerol was never depleted in Suspended-Cell reactors. Evidently, biofilm reactors present a reliable strategy to address the operational issues that occur during MK-7 biosynthesis on an industrial scale production.
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implementation of fed batch strategies for vitamin k menaquinone 7 production by bacillus subtilis natto in biofilm reactors
Applied Microbiology and Biotechnology, 2018Co-Authors: Ehsan Mahdinia, Ali Demirci, Aydin BerenjianAbstract:Recent studies show the essential health benefits associated with vitamin K, especially menaquinone-7 (MK-7). These benefits include reducing risks of cardiovascular diseases, osteoporosis, and even cancer. However, MK-7 production on an industrial level is only possible through bacterial fermentation and also current static fermentation strategies are not potent enough with difficulties to scale up. Biofilm reactors, however, may be a practical alternative. Biofilm reactors provide a controlled environment for the microorganisms to form mature and robust biofilms that enable them to produce value-added products with enhanced efficiencies. In this study, fed-batch addition of glucose and glycerol were investigated to the base media in biofilm reactors, as carbon source addition seemed crucial in batch fermentations. Results indicated that fed-batch strategies can be significantly effective in glucose-based medium, increasing the end-product concentrations to 28.7 ± 0.3 mg/L of MK-7 which was 2.3 fold higher than the level produced in Suspended-Cell bioreactors and renders the biofilm reactors as a potential replacement for static fermentation strategies. Moreover, morphological changes of B. subtilis were tracked during the 12-day long fermentation runs and finally, SEM investigations confirmed significant biofilm and extraCellular matrices formed on the plastic composite support (PCS) in the biofilm reactors. In conclusion, biofilm reactors especially with fed-batch fermentation regimes seem to be an effective tool for MK-7 production at industrial scales.
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optimization of bacillus subtilis natto growth parameters in glycerol based medium for vitamin k menaquinone 7 production in biofilm reactors
Bioprocess and Biosystems Engineering, 2018Co-Authors: Ehsan Mahdinia, Ali Demirci, Aydin BerenjianAbstract:Menaquinone-7 (MK-7) is the key form of vitamin K used as a dietary supplement and its production revolves around Bacillus subtilis natto. Current fermentation strategies, which suggest static fermentations without aeration and agitation, can be problematic for large scale MK-7 production due to biofilm formation. The use of biofilm reactors, therefore, is proposed in the present study, which could utilize both agitation and aeration without interrupting MK-7 secretion. In this study, biofilm reactors were constructed using the selected plastic composite support (PCS) and B. subtilis natto strain for MK-7 production. Using response surface methodology (RSM), optimum growth parameters including temperature, pH, and agitation were determined in a glycerol-based medium. Results were presented in a statistical model (R 2 = 0.90), leading to optimum growth conditions of temperature (35 °C), agitation (200 rpm) and pH (6.58). Model-predicted MK-7 concentration was validated and MK-7 concentration of 12.09 mg/L was produced in the biofilm reactor. The obtained concentration was 58% higher as compared to the Suspended-Cell culture (7.67 mg/L). The results of this study will provide a critical step towards improved industrial scale production of MK-7.
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utilization of glucose based medium and optimization of bacillus subtilis natto growth parameters for vitamin k menaquinone 7 production in biofilm reactors
Biocatalysis and agricultural biotechnology, 2018Co-Authors: Ehsan Mahdinia, Ali Demirci, Aydin BerenjianAbstract:Abstract Menaquinone-7 (MK-7) is the most potent form of vitamin K prescribed as a dietary supplement. MK-7 is commonly produced via static fermentation of Bacillus subtilis natto. The fermentation of this bacterium is associated with formation of large amounts of pellicles and biofilm, which are effective in MK-7 production, but also result in significant heat and mass transfer challenges during the scale-up process. Thus, the objectives of this study were to develop and evaluate the possibility of using a biofilm reactor for MK-7 production. In this research, biofilm reactors were constructed using the Plastic Composite Supports (PCS). In order to optimize the fermentation parameters for MK-7 production, Central Composite Design (CCD) was carried out along with supplementary runs to determine the optimum temperature, pH, and agitation. The biofilm reactors were able to produce a maximum concentration of 18.45 ± 0.76 mg/L of MK-7, which was 237% higher than the Suspended-Cell fermentation. Therefore, the present work suggests the possibility of using biofilm reactors as a new and effective fermentation strategy to address the issues associated with MK-7 fermentation.
Ali Demirci - One of the best experts on this subject based on the ideXlab platform.
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effects of medium components in a glycerol based medium on vitamin k menaquinone 7 production by bacillus subtilis natto in biofilm reactors
Bioprocess and Biosystems Engineering, 2019Co-Authors: Ehsan Mahdinia, Ali Demirci, Aydin BerenjianAbstract:Menaquinone-7 (MK-7) as the most important form of Vitamin K has been reported to have miraculous benefits such as preventing cardiovascular diseases and osteoporosis along with antitumor effects. Therefore, there have been numerous studies in the past decades to improve MK-7 production via microbial fermentation. Unfortunately, both solid and liquid state fermentation strategies that are utilized for MK-7 production, face fundamental operational and scale-up issues as well as intense heat and mass transfer problems during fermentation. In this regard, biofilm reactors seem to be a practical solution to overcome these issues and enhance the production in agitated liquid fermentation. Therefore, this study was undertaken to utilize biofilm reactors in investigating and optimizing different media components in a glycerol-based medium. Using response surface methodology, the effects of glycerol, yeast extract, and soytone were studied in the fermentation medium on MK-7 production in biofilm reactor. With a composition of 48.2 g/L of glycerol, 8.1 g/L of yeast extracts, 13.6 g/L of soytone and 0.06 g/L of K2HPO4, MK-7 concentrations could reach 14.7 ± 1.4 mg/L in biofilm reactors, which was 57% higher compared to the MK-7 concentration achieved in Suspended-Cell reactors under similar conditions, while glycerol was depleted by the end of the fifth day in biofilm reactors, but glycerol was never depleted in Suspended-Cell reactors. Evidently, biofilm reactors present a reliable strategy to address the operational issues that occur during MK-7 biosynthesis on an industrial scale production.
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implementation of fed batch strategies for vitamin k menaquinone 7 production by bacillus subtilis natto in biofilm reactors
Applied Microbiology and Biotechnology, 2018Co-Authors: Ehsan Mahdinia, Ali Demirci, Aydin BerenjianAbstract:Recent studies show the essential health benefits associated with vitamin K, especially menaquinone-7 (MK-7). These benefits include reducing risks of cardiovascular diseases, osteoporosis, and even cancer. However, MK-7 production on an industrial level is only possible through bacterial fermentation and also current static fermentation strategies are not potent enough with difficulties to scale up. Biofilm reactors, however, may be a practical alternative. Biofilm reactors provide a controlled environment for the microorganisms to form mature and robust biofilms that enable them to produce value-added products with enhanced efficiencies. In this study, fed-batch addition of glucose and glycerol were investigated to the base media in biofilm reactors, as carbon source addition seemed crucial in batch fermentations. Results indicated that fed-batch strategies can be significantly effective in glucose-based medium, increasing the end-product concentrations to 28.7 ± 0.3 mg/L of MK-7 which was 2.3 fold higher than the level produced in Suspended-Cell bioreactors and renders the biofilm reactors as a potential replacement for static fermentation strategies. Moreover, morphological changes of B. subtilis were tracked during the 12-day long fermentation runs and finally, SEM investigations confirmed significant biofilm and extraCellular matrices formed on the plastic composite support (PCS) in the biofilm reactors. In conclusion, biofilm reactors especially with fed-batch fermentation regimes seem to be an effective tool for MK-7 production at industrial scales.
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optimization of bacillus subtilis natto growth parameters in glycerol based medium for vitamin k menaquinone 7 production in biofilm reactors
Bioprocess and Biosystems Engineering, 2018Co-Authors: Ehsan Mahdinia, Ali Demirci, Aydin BerenjianAbstract:Menaquinone-7 (MK-7) is the key form of vitamin K used as a dietary supplement and its production revolves around Bacillus subtilis natto. Current fermentation strategies, which suggest static fermentations without aeration and agitation, can be problematic for large scale MK-7 production due to biofilm formation. The use of biofilm reactors, therefore, is proposed in the present study, which could utilize both agitation and aeration without interrupting MK-7 secretion. In this study, biofilm reactors were constructed using the selected plastic composite support (PCS) and B. subtilis natto strain for MK-7 production. Using response surface methodology (RSM), optimum growth parameters including temperature, pH, and agitation were determined in a glycerol-based medium. Results were presented in a statistical model (R 2 = 0.90), leading to optimum growth conditions of temperature (35 °C), agitation (200 rpm) and pH (6.58). Model-predicted MK-7 concentration was validated and MK-7 concentration of 12.09 mg/L was produced in the biofilm reactor. The obtained concentration was 58% higher as compared to the Suspended-Cell culture (7.67 mg/L). The results of this study will provide a critical step towards improved industrial scale production of MK-7.
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utilization of glucose based medium and optimization of bacillus subtilis natto growth parameters for vitamin k menaquinone 7 production in biofilm reactors
Biocatalysis and agricultural biotechnology, 2018Co-Authors: Ehsan Mahdinia, Ali Demirci, Aydin BerenjianAbstract:Abstract Menaquinone-7 (MK-7) is the most potent form of vitamin K prescribed as a dietary supplement. MK-7 is commonly produced via static fermentation of Bacillus subtilis natto. The fermentation of this bacterium is associated with formation of large amounts of pellicles and biofilm, which are effective in MK-7 production, but also result in significant heat and mass transfer challenges during the scale-up process. Thus, the objectives of this study were to develop and evaluate the possibility of using a biofilm reactor for MK-7 production. In this research, biofilm reactors were constructed using the Plastic Composite Supports (PCS). In order to optimize the fermentation parameters for MK-7 production, Central Composite Design (CCD) was carried out along with supplementary runs to determine the optimum temperature, pH, and agitation. The biofilm reactors were able to produce a maximum concentration of 18.45 ± 0.76 mg/L of MK-7, which was 237% higher than the Suspended-Cell fermentation. Therefore, the present work suggests the possibility of using biofilm reactors as a new and effective fermentation strategy to address the issues associated with MK-7 fermentation.
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Enhanced human lysozyme production in biofilm reactor by Kluyveromyces lactis K7
Biochemical Engineering Journal, 2014Co-Authors: Duygu Ercan, Ali DemirciAbstract:Abstract Lysozyme is an antimicrobial compound, which has been used in pharmaceutical and food industries. Chicken egg is the commercial source of lysozyme. However, human lysozyme is more effective and safer than egg-white lysozyme. Human milk is an important source for human lysozyme, but it is not feasible to provide the needed lysozyme commercially. Biofilm reactors provide passive immobilization of Cells onto the solid support, which may lead to higher productivity. The aim was to evaluate the fermentation medium composition for enhanced human lysozyme production by Kluyveromyces lactis K7 in biofilm reactor with plastic composite supports. Yeast nitrogen base was selected as the best nitrogen source when compared to the yeast extract and corn steep liquor. Moreover, inhibition effect of NaCl and NH4Cl at the concentrations of 25 and 50 mM was observed. Three factors Box–Behnken response surface design was conducted and the results suggested 16.3% lactose, 1.2% casamino acid, 0.8% yeast nitrogen base as optimum medium composition for maximum human lysozyme production. Overall, the human lysozyme production by K. lactis K7 was increased to 173 U/ml, which is about 23% improvement in biofilm reactor and 57% improvement compared to the Suspended-Cell fermentation.
Ehsan Mahdinia - One of the best experts on this subject based on the ideXlab platform.
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effects of medium components in a glycerol based medium on vitamin k menaquinone 7 production by bacillus subtilis natto in biofilm reactors
Bioprocess and Biosystems Engineering, 2019Co-Authors: Ehsan Mahdinia, Ali Demirci, Aydin BerenjianAbstract:Menaquinone-7 (MK-7) as the most important form of Vitamin K has been reported to have miraculous benefits such as preventing cardiovascular diseases and osteoporosis along with antitumor effects. Therefore, there have been numerous studies in the past decades to improve MK-7 production via microbial fermentation. Unfortunately, both solid and liquid state fermentation strategies that are utilized for MK-7 production, face fundamental operational and scale-up issues as well as intense heat and mass transfer problems during fermentation. In this regard, biofilm reactors seem to be a practical solution to overcome these issues and enhance the production in agitated liquid fermentation. Therefore, this study was undertaken to utilize biofilm reactors in investigating and optimizing different media components in a glycerol-based medium. Using response surface methodology, the effects of glycerol, yeast extract, and soytone were studied in the fermentation medium on MK-7 production in biofilm reactor. With a composition of 48.2 g/L of glycerol, 8.1 g/L of yeast extracts, 13.6 g/L of soytone and 0.06 g/L of K2HPO4, MK-7 concentrations could reach 14.7 ± 1.4 mg/L in biofilm reactors, which was 57% higher compared to the MK-7 concentration achieved in Suspended-Cell reactors under similar conditions, while glycerol was depleted by the end of the fifth day in biofilm reactors, but glycerol was never depleted in Suspended-Cell reactors. Evidently, biofilm reactors present a reliable strategy to address the operational issues that occur during MK-7 biosynthesis on an industrial scale production.
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implementation of fed batch strategies for vitamin k menaquinone 7 production by bacillus subtilis natto in biofilm reactors
Applied Microbiology and Biotechnology, 2018Co-Authors: Ehsan Mahdinia, Ali Demirci, Aydin BerenjianAbstract:Recent studies show the essential health benefits associated with vitamin K, especially menaquinone-7 (MK-7). These benefits include reducing risks of cardiovascular diseases, osteoporosis, and even cancer. However, MK-7 production on an industrial level is only possible through bacterial fermentation and also current static fermentation strategies are not potent enough with difficulties to scale up. Biofilm reactors, however, may be a practical alternative. Biofilm reactors provide a controlled environment for the microorganisms to form mature and robust biofilms that enable them to produce value-added products with enhanced efficiencies. In this study, fed-batch addition of glucose and glycerol were investigated to the base media in biofilm reactors, as carbon source addition seemed crucial in batch fermentations. Results indicated that fed-batch strategies can be significantly effective in glucose-based medium, increasing the end-product concentrations to 28.7 ± 0.3 mg/L of MK-7 which was 2.3 fold higher than the level produced in Suspended-Cell bioreactors and renders the biofilm reactors as a potential replacement for static fermentation strategies. Moreover, morphological changes of B. subtilis were tracked during the 12-day long fermentation runs and finally, SEM investigations confirmed significant biofilm and extraCellular matrices formed on the plastic composite support (PCS) in the biofilm reactors. In conclusion, biofilm reactors especially with fed-batch fermentation regimes seem to be an effective tool for MK-7 production at industrial scales.
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optimization of bacillus subtilis natto growth parameters in glycerol based medium for vitamin k menaquinone 7 production in biofilm reactors
Bioprocess and Biosystems Engineering, 2018Co-Authors: Ehsan Mahdinia, Ali Demirci, Aydin BerenjianAbstract:Menaquinone-7 (MK-7) is the key form of vitamin K used as a dietary supplement and its production revolves around Bacillus subtilis natto. Current fermentation strategies, which suggest static fermentations without aeration and agitation, can be problematic for large scale MK-7 production due to biofilm formation. The use of biofilm reactors, therefore, is proposed in the present study, which could utilize both agitation and aeration without interrupting MK-7 secretion. In this study, biofilm reactors were constructed using the selected plastic composite support (PCS) and B. subtilis natto strain for MK-7 production. Using response surface methodology (RSM), optimum growth parameters including temperature, pH, and agitation were determined in a glycerol-based medium. Results were presented in a statistical model (R 2 = 0.90), leading to optimum growth conditions of temperature (35 °C), agitation (200 rpm) and pH (6.58). Model-predicted MK-7 concentration was validated and MK-7 concentration of 12.09 mg/L was produced in the biofilm reactor. The obtained concentration was 58% higher as compared to the Suspended-Cell culture (7.67 mg/L). The results of this study will provide a critical step towards improved industrial scale production of MK-7.
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utilization of glucose based medium and optimization of bacillus subtilis natto growth parameters for vitamin k menaquinone 7 production in biofilm reactors
Biocatalysis and agricultural biotechnology, 2018Co-Authors: Ehsan Mahdinia, Ali Demirci, Aydin BerenjianAbstract:Abstract Menaquinone-7 (MK-7) is the most potent form of vitamin K prescribed as a dietary supplement. MK-7 is commonly produced via static fermentation of Bacillus subtilis natto. The fermentation of this bacterium is associated with formation of large amounts of pellicles and biofilm, which are effective in MK-7 production, but also result in significant heat and mass transfer challenges during the scale-up process. Thus, the objectives of this study were to develop and evaluate the possibility of using a biofilm reactor for MK-7 production. In this research, biofilm reactors were constructed using the Plastic Composite Supports (PCS). In order to optimize the fermentation parameters for MK-7 production, Central Composite Design (CCD) was carried out along with supplementary runs to determine the optimum temperature, pH, and agitation. The biofilm reactors were able to produce a maximum concentration of 18.45 ± 0.76 mg/L of MK-7, which was 237% higher than the Suspended-Cell fermentation. Therefore, the present work suggests the possibility of using biofilm reactors as a new and effective fermentation strategy to address the issues associated with MK-7 fermentation.
Dario Frascari - One of the best experts on this subject based on the ideXlab platform.
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Aerobic cometabolism of 1,1,2,2-tetrachloroethane by Rhodococcus aetherivorans TPA grown on propane: kinetic study and bioreactor configuration analysis
Journal of Chemical Technology & Biotechnology, 2017Co-Authors: Martina Cappelletti, Davide Pinelli, Stefano Fedi, Davide Zannoni, Dario FrascariAbstract:BACKGROUND 1,1,2,2-Tetrachloroethane (TeCA) has been generally considered as non-biodegradable under aerobic conditions, while its complete biodegradation was reported with microbial consortia growing anaerobically. This study describes TeCA aerobic cometabolic degradation by the propanotroph Rhodococcus aetherivorans strain TPA isolated from a TeCA-degrading consortium. RESULTS R. aetherivorans TPA was able to grow on aliphatic hydrocarbons from propane to pentane and on gaseous n-alkanes metabolic intermediates. The Michaelis–Menten model allowed a satisfactory fit of the TPA propane utilization rates under resting Cell conditions, while the TeCA degradation rates were successfully interpolated with Andrew's inhibition model. A significant propane - TeCA mutual inhibition was observed, although the results did not allow to distinguish between competitive and non-competitive inhibition. Among different bioreactor options for the on-site bioremediation of TeCA-contaminated groundwater, a single Suspended-Cell continuous stirred-tank reactor (CSTR) appeared to be the optimal one. CONCLUSIONS This study provides for the first time the kinetic and microbiological characterization of a bacterial strain capable to degrade TeCA under aerobic conditions.
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aerobic cometabolism of 1 1 2 2 teca by a propane growing microbial consortium c2 diversity of alkane monooxygenase genes and design of an on site bioremediation process
International Biodeterioration & Biodegradation, 2017Co-Authors: Martina Cappelletti, Davide Pinelli, Stefano Fedi, Dario Frascari, Francesco Mezzetti, Davide ZannoniAbstract:Microbial degradation of 1,1,2,2-tetrachloroethane has been rarely analysed under aerobic conditions. In this work, the catabolic potential of a TeCA-degrading aerobic propanotroph consortium (C2) and the optimal bioreactor configuration for an on-site TeCA-bioremediation strategy with C2 were defined. More specifically, the diversity of alkane-oxidizing bacteria in C2 was assessed by means of clone libraries of genes coding for alkane monooxygenases (MOs) of different families (AlkB-like alkane hydroxylase, soluble di-iron MO and cytochromes P450). A large number of alkane MO sequences retrieved in this study showed the highest similarity with reference sequences belonging to Rhodococcus genus, suggesting a key role of this genus in TeCA/propane co-metabolism, while the remaining alkane MO sequences were mainly attributed to other Actinobacteria, to Bradhyrizobiaceae, and Cupriavidus genus. Further, the feasibility of an on-site TeCA bioremediation strategy with C2 was evaluated by simulating a continuous-flow aerobic co-metabolic process with different bioreactor configurations. Our results show that the configuration with a Suspended-Cell continuous stirred-tank reactor (CSTR) followed by a Suspended-Cell plug-flow reactor (PFR) was the one giving the best performance with consortium C2.
Martina Cappelletti - One of the best experts on this subject based on the ideXlab platform.
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Aerobic cometabolism of 1,1,2,2-tetrachloroethane by Rhodococcus aetherivorans TPA grown on propane: kinetic study and bioreactor configuration analysis
Journal of Chemical Technology & Biotechnology, 2017Co-Authors: Martina Cappelletti, Davide Pinelli, Stefano Fedi, Davide Zannoni, Dario FrascariAbstract:BACKGROUND 1,1,2,2-Tetrachloroethane (TeCA) has been generally considered as non-biodegradable under aerobic conditions, while its complete biodegradation was reported with microbial consortia growing anaerobically. This study describes TeCA aerobic cometabolic degradation by the propanotroph Rhodococcus aetherivorans strain TPA isolated from a TeCA-degrading consortium. RESULTS R. aetherivorans TPA was able to grow on aliphatic hydrocarbons from propane to pentane and on gaseous n-alkanes metabolic intermediates. The Michaelis–Menten model allowed a satisfactory fit of the TPA propane utilization rates under resting Cell conditions, while the TeCA degradation rates were successfully interpolated with Andrew's inhibition model. A significant propane - TeCA mutual inhibition was observed, although the results did not allow to distinguish between competitive and non-competitive inhibition. Among different bioreactor options for the on-site bioremediation of TeCA-contaminated groundwater, a single Suspended-Cell continuous stirred-tank reactor (CSTR) appeared to be the optimal one. CONCLUSIONS This study provides for the first time the kinetic and microbiological characterization of a bacterial strain capable to degrade TeCA under aerobic conditions.
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aerobic cometabolism of 1 1 2 2 teca by a propane growing microbial consortium c2 diversity of alkane monooxygenase genes and design of an on site bioremediation process
International Biodeterioration & Biodegradation, 2017Co-Authors: Martina Cappelletti, Davide Pinelli, Stefano Fedi, Dario Frascari, Francesco Mezzetti, Davide ZannoniAbstract:Microbial degradation of 1,1,2,2-tetrachloroethane has been rarely analysed under aerobic conditions. In this work, the catabolic potential of a TeCA-degrading aerobic propanotroph consortium (C2) and the optimal bioreactor configuration for an on-site TeCA-bioremediation strategy with C2 were defined. More specifically, the diversity of alkane-oxidizing bacteria in C2 was assessed by means of clone libraries of genes coding for alkane monooxygenases (MOs) of different families (AlkB-like alkane hydroxylase, soluble di-iron MO and cytochromes P450). A large number of alkane MO sequences retrieved in this study showed the highest similarity with reference sequences belonging to Rhodococcus genus, suggesting a key role of this genus in TeCA/propane co-metabolism, while the remaining alkane MO sequences were mainly attributed to other Actinobacteria, to Bradhyrizobiaceae, and Cupriavidus genus. Further, the feasibility of an on-site TeCA bioremediation strategy with C2 was evaluated by simulating a continuous-flow aerobic co-metabolic process with different bioreactor configurations. Our results show that the configuration with a Suspended-Cell continuous stirred-tank reactor (CSTR) followed by a Suspended-Cell plug-flow reactor (PFR) was the one giving the best performance with consortium C2.
