Improving the bioremediation of phenolic wastewaters by Trametes versicolor
- Ryan, D, Leukes, Winston D, Burton, Stephanie G
- Authors: Ryan, D , Leukes, Winston D , Burton, Stephanie G
- Date: 2006
- Language: English
- Type: Article
- Identifier: vital:6467 , http://hdl.handle.net/10962/d1005796 , http://dx.doi.org/10.1016/j.biortech.2006.02.001
- Description: The successful bioremediation of a phenolic wastewater by Trametes versicolor was found to be dependent on a range of factors including: fungal growth, culture age and activity and enzyme (laccase) production. These aspects were enhanced by the optimisation of the growth medium used and time of addition of the pollutant to the fungal cultures. Different media containing ‘high’ (20 g/L), ‘low’ (2 g/L) and ‘sufficient’ (10 g/L) concentrations of carbon and nitrogen sources were investigated. The medium containing both glucose and peptone at 10 g/L resulted in the highest Growth Related Productivity (the product of specific yield and μ) of laccase (1.46 Units of laccase activity)/gram biomass/day and was used in all further experiments. The use of the guaiacol as an inducer further increased laccase activity 780% without inhibiting growth; similarly the phenolic effluent studied boosted activity almost 5 times. The timing of the addition of the phenolic effluent was found to have important consequences in its removal and at least 8 days of prior growth was required. Under these conditions, 0.125 g phenol/g biomass and 0.231 g o-cresol/g biomass were removed from solution per day.
- Full Text:
- Date Issued: 2006
- Authors: Ryan, D , Leukes, Winston D , Burton, Stephanie G
- Date: 2006
- Language: English
- Type: Article
- Identifier: vital:6467 , http://hdl.handle.net/10962/d1005796 , http://dx.doi.org/10.1016/j.biortech.2006.02.001
- Description: The successful bioremediation of a phenolic wastewater by Trametes versicolor was found to be dependent on a range of factors including: fungal growth, culture age and activity and enzyme (laccase) production. These aspects were enhanced by the optimisation of the growth medium used and time of addition of the pollutant to the fungal cultures. Different media containing ‘high’ (20 g/L), ‘low’ (2 g/L) and ‘sufficient’ (10 g/L) concentrations of carbon and nitrogen sources were investigated. The medium containing both glucose and peptone at 10 g/L resulted in the highest Growth Related Productivity (the product of specific yield and μ) of laccase (1.46 Units of laccase activity)/gram biomass/day and was used in all further experiments. The use of the guaiacol as an inducer further increased laccase activity 780% without inhibiting growth; similarly the phenolic effluent studied boosted activity almost 5 times. The timing of the addition of the phenolic effluent was found to have important consequences in its removal and at least 8 days of prior growth was required. Under these conditions, 0.125 g phenol/g biomass and 0.231 g o-cresol/g biomass were removed from solution per day.
- Full Text:
- Date Issued: 2006
Competition for attachment of aquaculture candidate probiotic and pathogenic bacteria on fish intestinal mucus:
- Vine, Niall G, Leukes, Winston D, Kaiser, Horst, Daya, Santylal, Baxter, Jeremy, Hecht, Thomas
- Authors: Vine, Niall G , Leukes, Winston D , Kaiser, Horst , Daya, Santylal , Baxter, Jeremy , Hecht, Thomas
- Date: 2004
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/142819 , vital:38120 , DOI: 10.1111/j.1365-2761.2004.00542.x
- Description: Probiotics for aquaculture are generally only selected by their ability to produce antimicrobial metabolites; however, attachment to intestinal mucus is important in order to remain within the gut of its host. Five candidate probiotics (AP1–AP5), isolated from the clownfish, Amphiprion percula (Lacepéde), were examined for their ability to attach to fish intestinal mucus and compete with two pathogens, Aeromonas hydrophila and Vibrio alginolyticus. Two different radioactive isotopes were used to quantify competition between pathogens and probionts. Attachment of the pathogens was enhanced by the presence of the candidate probiotics. However, the addition of the candidate probiotics after the pathogens resulted in reduced pathogen attachment.
- Full Text:
- Date Issued: 2004
- Authors: Vine, Niall G , Leukes, Winston D , Kaiser, Horst , Daya, Santylal , Baxter, Jeremy , Hecht, Thomas
- Date: 2004
- Language: English
- Type: text , article
- Identifier: http://hdl.handle.net/10962/142819 , vital:38120 , DOI: 10.1111/j.1365-2761.2004.00542.x
- Description: Probiotics for aquaculture are generally only selected by their ability to produce antimicrobial metabolites; however, attachment to intestinal mucus is important in order to remain within the gut of its host. Five candidate probiotics (AP1–AP5), isolated from the clownfish, Amphiprion percula (Lacepéde), were examined for their ability to attach to fish intestinal mucus and compete with two pathogens, Aeromonas hydrophila and Vibrio alginolyticus. Two different radioactive isotopes were used to quantify competition between pathogens and probionts. Attachment of the pathogens was enhanced by the presence of the candidate probiotics. However, the addition of the candidate probiotics after the pathogens resulted in reduced pathogen attachment.
- Full Text:
- Date Issued: 2004
Cleaning fouled membranes using sludge enzymes
- Melamane, Xolisa L, Pletschke, Brett I, Leukes, Winston D, Whiteley, Chris G
- Authors: Melamane, Xolisa L , Pletschke, Brett I , Leukes, Winston D , Whiteley, Chris G
- Date: 2003
- Language: English
- Type: text
- Identifier: vital:6480 , http://hdl.handle.net/10962/d1006242
- Description: Maintenance of membrane performance requires inevitable cleaning or "defouling" of fouled membranes. Membrane cleaning using sludge enzymes, was investigated by first characterising ostrich abattoir effluent for potential foulants, such as lipids, proteins and polysaccharides. Static fouling of polysulphone membranes using abattoir effluent was also performed. Biochemical analysis was performed using quantitative and qualitative methods for detection of proteins on fouled and defouled membranes. The ability of sulphidogenic proteases to remove proteins adsorbed on polysulphone membranes and capillary ultrafiltration membranes after static fouling, and ability to restore permeate fluxes and transmembrane pressure after dynamic fouling was also investigated. Permeate volumes were analysed for protein and amino acids concentrations. The abattoir effluent contained 553 μg/ml of lipid, 301 μg/ml of protein, 141 μg/ml of total carbohydrate, and 0.63 μg/ml of total reducing sugars. Static fouled membranes removed 23.4percent of proteins. Defouling of dynamically fouled capillary ultrafiltration membranes using sulphidogenic proteases was successful at pH 10, 37°C, within 1 h. Sulphidogenic protease activity was 2.1 U/ml and Flux Recovery (FR percent) was 64 percent.
- Full Text:
- Date Issued: 2003
- Authors: Melamane, Xolisa L , Pletschke, Brett I , Leukes, Winston D , Whiteley, Chris G
- Date: 2003
- Language: English
- Type: text
- Identifier: vital:6480 , http://hdl.handle.net/10962/d1006242
- Description: Maintenance of membrane performance requires inevitable cleaning or "defouling" of fouled membranes. Membrane cleaning using sludge enzymes, was investigated by first characterising ostrich abattoir effluent for potential foulants, such as lipids, proteins and polysaccharides. Static fouling of polysulphone membranes using abattoir effluent was also performed. Biochemical analysis was performed using quantitative and qualitative methods for detection of proteins on fouled and defouled membranes. The ability of sulphidogenic proteases to remove proteins adsorbed on polysulphone membranes and capillary ultrafiltration membranes after static fouling, and ability to restore permeate fluxes and transmembrane pressure after dynamic fouling was also investigated. Permeate volumes were analysed for protein and amino acids concentrations. The abattoir effluent contained 553 μg/ml of lipid, 301 μg/ml of protein, 141 μg/ml of total carbohydrate, and 0.63 μg/ml of total reducing sugars. Static fouled membranes removed 23.4percent of proteins. Defouling of dynamically fouled capillary ultrafiltration membranes using sulphidogenic proteases was successful at pH 10, 37°C, within 1 h. Sulphidogenic protease activity was 2.1 U/ml and Flux Recovery (FR percent) was 64 percent.
- Full Text:
- Date Issued: 2003
Physical and biological variability in the Antarctic Polar Frontal Zone: report on research cruise 103 of the MV SA Agulhas
- Froneman, P William, Ansorge, Isabelle J, Vumazonke, Lukhanyiso U, Gulekana, A, Bernard, Kim S, Webb, Arthur C M, Leukes, Winston D, Risien, C M, Thomalla, S, Hermes, Juliet, Knott, M, Anderson, D, Hargey, N, Jennings, Michael E, Veitch, J, Lutjeharms, Johan R E, McQuaid, Christopher D
- Authors: Froneman, P William , Ansorge, Isabelle J , Vumazonke, Lukhanyiso U , Gulekana, A , Bernard, Kim S , Webb, Arthur C M , Leukes, Winston D , Risien, C M , Thomalla, S , Hermes, Juliet , Knott, M , Anderson, D , Hargey, N , Jennings, Michael E , Veitch, J , Lutjeharms, Johan R E , McQuaid, Christopher D
- Date: 2002
- Language: English
- Type: Article
- Identifier: vital:6910 , http://hdl.handle.net/10962/d1011863
- Description: A detailed hydrographic and biological survey was carried out in the region of the South-west Indian Ridge during April 2002. Hydrographic data revealed that the Andrew Bain Fracture Zone, centred at 30oE, 50oS, functions as an important choke point to the flow of the Antarctic Circumpolar Current, resulting in the convergence of the Antarctic Polar Front (APF) and the southern branch of the Sub-Antarctic Front (SSAF). Total chlorophyll-a concentration and zooplankton biomass were highest at stations occupied in the vicinity of two frontal features represented by the APF and SSAF. These data suggest that the region of the South-west Indian Ridge is an area of elevated biological activity and probably acts as an important offshore feeding area for the top predators on the Prince Edward Islands.
- Full Text:
- Date Issued: 2002
- Authors: Froneman, P William , Ansorge, Isabelle J , Vumazonke, Lukhanyiso U , Gulekana, A , Bernard, Kim S , Webb, Arthur C M , Leukes, Winston D , Risien, C M , Thomalla, S , Hermes, Juliet , Knott, M , Anderson, D , Hargey, N , Jennings, Michael E , Veitch, J , Lutjeharms, Johan R E , McQuaid, Christopher D
- Date: 2002
- Language: English
- Type: Article
- Identifier: vital:6910 , http://hdl.handle.net/10962/d1011863
- Description: A detailed hydrographic and biological survey was carried out in the region of the South-west Indian Ridge during April 2002. Hydrographic data revealed that the Andrew Bain Fracture Zone, centred at 30oE, 50oS, functions as an important choke point to the flow of the Antarctic Circumpolar Current, resulting in the convergence of the Antarctic Polar Front (APF) and the southern branch of the Sub-Antarctic Front (SSAF). Total chlorophyll-a concentration and zooplankton biomass were highest at stations occupied in the vicinity of two frontal features represented by the APF and SSAF. These data suggest that the region of the South-west Indian Ridge is an area of elevated biological activity and probably acts as an important offshore feeding area for the top predators on the Prince Edward Islands.
- Full Text:
- Date Issued: 2002
Development and characterisation of a membrane gradostat bioreactor for the bioremediation of aromatic pollutants using white rot fungi
- Authors: Leukes, Winston D
- Date: 1999
- Subjects: Aromatic compounds Pollutants Fungi Bioremediation Industrial microbiology Biotechnology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4032 , http://hdl.handle.net/10962/d1004092
- Description: Bioremediation of aromatic pollutants using the ligninolytic enzymes of the white rot fungi has been thoroughly researched and has been shown to have considerable potential for industrial application. However, little success in scale-up and industrialisation of this technology has been attained due to problems associated with the continuous production of the pollutant-degrading enzymes using conventional bioreactor systems. The low productivities reported result from the incompatibility of conventional submerged culture reactor techniques with the physiological requirements of these fungi which have evolved on a solid-air interface, viz. wood. The enzymes are also produced only during the stationary phase of growth and can therefore be regarded as secondary metabolites. This study reports the conceptualisation, characterisation and evaluation of a novel bioreactor system as a solution to the continuous production of idiophasic pollutant degrading enzymes by the white rot fungus Phanerochaete chlysosporium. The reactor concept evolved from observation of these fungi in their native state, i. e. the metabolism of lignocellulosic material and involves the immobilisation of the organism onto a capillary ultrafiltration membrane. Nutrient gradients established across the biofilm, an inherent characteristic of fixed bed perfusion reactors, are exploited to provide both nutrient rich and nutrient poor zones across the biofilm. This allows growth or primary metabolism in the nutrient rich zone, pushing older biomass into the nutrient poor zone where secondary metabolism is induced by nutrient starvation. In effect, this represents a transformation of the events of a batch culture from a temporal to a spatial domain, allowing continuous production of secondary metabolites over time. Direct contact of the outer part of the biofilm with an air stream simulated the solid-air interface of the native state of the fungus. In order to facilitate the practical application of the membrane gradostat reactor (MGR) concept, conventional capillary membranes and membrane bioreactor modules were first evaluated. These were found to be unsuitable for application of the MGR concept. However, critical analysis of the shortcomings of the conventional systems resulted in the formulation of a set of design criteria for the development of a suitable membrane and module. These design criteria were satisfied by the development of a novel capillary membrane for membrane bioreactors, as well as a transverse flow membrane module, which is a novel approach in membrane bioreactor configuration. For the physiological characterisation of the MGR concept, a single fibre bioreactor unit was designed, which allowed destructive sampling of the biofilm for analysis. Using this system, it was shown that distinct morphological zones could be observed radially across the mature biofilm obtained through MGR operation. That these morphotypes do represent the temporal events of a typical batch culture in a spatial domain was confirmed by following the morphological changes occurring during batch culture of the immobilised fungus where the onset of primary and secondary metabolic conditions were manipulated through control of the nutrient supply. The different morphotypes were correlated to distinct growth phases by comparison of the morphology to the secretion of known enzymatic markers for secondary metabolism, viz. succinate dehydrogenase and cytochrome C oxidoreductase. Detailed structure-function analysis of the biofilm using transmission electron microscopy and adapted enzyme cytochemical staining techniques showed that the biofilm appeared to operate as a co-ordinated unit, with primary and secondary metabolism apparently linked in one thallus through nutrient translocation. This study provided new insights into the physiology of P. chrysosp,o rium and a detailed descriptive model was formulated which correlates well to existing models of wood degradation by the white rot fungi (WRF). Evaluation of the process on a laboratory scale using a novel transverse flow membrane bioreactor showed that a volumetric productivity of 1916 U.L.⁻¹day⁻¹ for manganese peroxidase, one of the pollutant degrading enzymes, could be attained, corresponding to a final concentration of 2 361 U.L.⁻¹ This may be compared to the best reported system (Moreira el at. 1997), where a volumetric productivity of 202 U.L.⁻¹day⁻¹was achieved with a final concentration of 250 U.L.⁻¹ However, MGR productivity is yet to be subjected to rigorous optimisation studies. The process could be operated continuously for 60 days. However, peak productivity could not be maintained for long periods. This was found to be due to physical phenomena relating to the fluid dynamics of the system which caused fluid flow maldistribution, which would have to be resolved through engineering analysis. In evaluation of the MGR concept for aromatic pollutant removal, in this case ρ- cresol, from growth medium, good performance was also achieved. The VmaxKm calculated by linear regression for the MGR was 0.8 (R² = 0.93), which compared favourably to that reported by Lewandowski et al. (1990), who obtained a Vmax/Km of 0.34 for a packed bed reactor treating chlorophenol. It was concluded that the MGR showed suitable potential to warrant further development, and that the descriptive characterisation of the biofilm physiology provided a sufficient basis for process analysis once engineering aspects ofthe system could be resolved.
- Full Text:
- Date Issued: 1999
- Authors: Leukes, Winston D
- Date: 1999
- Subjects: Aromatic compounds Pollutants Fungi Bioremediation Industrial microbiology Biotechnology
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4032 , http://hdl.handle.net/10962/d1004092
- Description: Bioremediation of aromatic pollutants using the ligninolytic enzymes of the white rot fungi has been thoroughly researched and has been shown to have considerable potential for industrial application. However, little success in scale-up and industrialisation of this technology has been attained due to problems associated with the continuous production of the pollutant-degrading enzymes using conventional bioreactor systems. The low productivities reported result from the incompatibility of conventional submerged culture reactor techniques with the physiological requirements of these fungi which have evolved on a solid-air interface, viz. wood. The enzymes are also produced only during the stationary phase of growth and can therefore be regarded as secondary metabolites. This study reports the conceptualisation, characterisation and evaluation of a novel bioreactor system as a solution to the continuous production of idiophasic pollutant degrading enzymes by the white rot fungus Phanerochaete chlysosporium. The reactor concept evolved from observation of these fungi in their native state, i. e. the metabolism of lignocellulosic material and involves the immobilisation of the organism onto a capillary ultrafiltration membrane. Nutrient gradients established across the biofilm, an inherent characteristic of fixed bed perfusion reactors, are exploited to provide both nutrient rich and nutrient poor zones across the biofilm. This allows growth or primary metabolism in the nutrient rich zone, pushing older biomass into the nutrient poor zone where secondary metabolism is induced by nutrient starvation. In effect, this represents a transformation of the events of a batch culture from a temporal to a spatial domain, allowing continuous production of secondary metabolites over time. Direct contact of the outer part of the biofilm with an air stream simulated the solid-air interface of the native state of the fungus. In order to facilitate the practical application of the membrane gradostat reactor (MGR) concept, conventional capillary membranes and membrane bioreactor modules were first evaluated. These were found to be unsuitable for application of the MGR concept. However, critical analysis of the shortcomings of the conventional systems resulted in the formulation of a set of design criteria for the development of a suitable membrane and module. These design criteria were satisfied by the development of a novel capillary membrane for membrane bioreactors, as well as a transverse flow membrane module, which is a novel approach in membrane bioreactor configuration. For the physiological characterisation of the MGR concept, a single fibre bioreactor unit was designed, which allowed destructive sampling of the biofilm for analysis. Using this system, it was shown that distinct morphological zones could be observed radially across the mature biofilm obtained through MGR operation. That these morphotypes do represent the temporal events of a typical batch culture in a spatial domain was confirmed by following the morphological changes occurring during batch culture of the immobilised fungus where the onset of primary and secondary metabolic conditions were manipulated through control of the nutrient supply. The different morphotypes were correlated to distinct growth phases by comparison of the morphology to the secretion of known enzymatic markers for secondary metabolism, viz. succinate dehydrogenase and cytochrome C oxidoreductase. Detailed structure-function analysis of the biofilm using transmission electron microscopy and adapted enzyme cytochemical staining techniques showed that the biofilm appeared to operate as a co-ordinated unit, with primary and secondary metabolism apparently linked in one thallus through nutrient translocation. This study provided new insights into the physiology of P. chrysosp,o rium and a detailed descriptive model was formulated which correlates well to existing models of wood degradation by the white rot fungi (WRF). Evaluation of the process on a laboratory scale using a novel transverse flow membrane bioreactor showed that a volumetric productivity of 1916 U.L.⁻¹day⁻¹ for manganese peroxidase, one of the pollutant degrading enzymes, could be attained, corresponding to a final concentration of 2 361 U.L.⁻¹ This may be compared to the best reported system (Moreira el at. 1997), where a volumetric productivity of 202 U.L.⁻¹day⁻¹was achieved with a final concentration of 250 U.L.⁻¹ However, MGR productivity is yet to be subjected to rigorous optimisation studies. The process could be operated continuously for 60 days. However, peak productivity could not be maintained for long periods. This was found to be due to physical phenomena relating to the fluid dynamics of the system which caused fluid flow maldistribution, which would have to be resolved through engineering analysis. In evaluation of the MGR concept for aromatic pollutant removal, in this case ρ- cresol, from growth medium, good performance was also achieved. The VmaxKm calculated by linear regression for the MGR was 0.8 (R² = 0.93), which compared favourably to that reported by Lewandowski et al. (1990), who obtained a Vmax/Km of 0.34 for a packed bed reactor treating chlorophenol. It was concluded that the MGR showed suitable potential to warrant further development, and that the descriptive characterisation of the biofilm physiology provided a sufficient basis for process analysis once engineering aspects ofthe system could be resolved.
- Full Text:
- Date Issued: 1999
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