Environmental drivers of the composition and distribution of larval fish assemblages off the south coast of South Africa
- Authors: Trassierra, Jaqueline Anne
- Date: 2019
- Subjects: Fishes -- Larvae -- South Africa -- Eastern Cape , Fishes -- Larvae -- Migration -- South Africa -- Eastern Cape , Fishes -- Larvae -- Dispersal -- South Africa -- Eastern Cape
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/68154 , vital:29207
- Description: The species composition, distribution and patterns of vertical migration of larval fish assemblages were investigated in March and in September 2013 within two adjacent log spiral bays, Algoa Bay and St Francis Bay, on the south coast of South Africa. Fish larvae were collected by means of a boat towed bongo net (57 cm diameter; mesh aperture 500 μm). An onshore (2 km) and an offshore (3 km) station were each sampled twice during the daytime (06:00 – 18:00) with two horizontal tows: near the surface (0.5 m) and close to the bottom (12 m). Tows were repeated at night (18:00 – 23:00) for onshore sites. Larval catches included 16 fish families and 40 species. A multivariate analysis indicated that the species composition was significantly different between Algoa Bay and St Francis Bay, with Engraulidae, Blenniidae, Sparidae, Soleidae and Cynoglossidae making important contributions to the larval fish catch in Algoa Bay, while Blenniidae, Engraulidae, Tripterygiidae, Sparidae and Gobiesocidae contributed significantly in St Francis Bay. Differences in assemblage composition were noted between the Spring (September–October) and Autumn (March-April) months. The species composition of larval fish assemblages was related to wind speed, wave height, cloud cover, sea water temperature, depth, average current speed and direction. Wind speed, wave height, temperature and depth significantly contributed to the variation in larval fish densities. Abundances of larval fishes were greater offshore than onshore, larvae from pelagic eggs dominated catches offshore, while larvae from demersal eggs dominated onshore catches. Habitat structure strongly influenced the composition of larval fishes between the bays and abundances were significantly greater at night than during the day. Most larval fishes displayed a reverse diel vertical migration pattern and were most influenced by predators, wind speed and cloud cover. This study shows that larval fish assemblages are highly complex and patchy. Spawning mode, individual species behaviour, diel vertical migration, current structure, depth, temperature, wind speed, cloud cover and type of habitat substratum all influence larval fish composition and distribution in the nearshore waters of South Africa.
- Full Text:
- Date Issued: 2019
- Authors: Trassierra, Jaqueline Anne
- Date: 2019
- Subjects: Fishes -- Larvae -- South Africa -- Eastern Cape , Fishes -- Larvae -- Migration -- South Africa -- Eastern Cape , Fishes -- Larvae -- Dispersal -- South Africa -- Eastern Cape
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/68154 , vital:29207
- Description: The species composition, distribution and patterns of vertical migration of larval fish assemblages were investigated in March and in September 2013 within two adjacent log spiral bays, Algoa Bay and St Francis Bay, on the south coast of South Africa. Fish larvae were collected by means of a boat towed bongo net (57 cm diameter; mesh aperture 500 μm). An onshore (2 km) and an offshore (3 km) station were each sampled twice during the daytime (06:00 – 18:00) with two horizontal tows: near the surface (0.5 m) and close to the bottom (12 m). Tows were repeated at night (18:00 – 23:00) for onshore sites. Larval catches included 16 fish families and 40 species. A multivariate analysis indicated that the species composition was significantly different between Algoa Bay and St Francis Bay, with Engraulidae, Blenniidae, Sparidae, Soleidae and Cynoglossidae making important contributions to the larval fish catch in Algoa Bay, while Blenniidae, Engraulidae, Tripterygiidae, Sparidae and Gobiesocidae contributed significantly in St Francis Bay. Differences in assemblage composition were noted between the Spring (September–October) and Autumn (March-April) months. The species composition of larval fish assemblages was related to wind speed, wave height, cloud cover, sea water temperature, depth, average current speed and direction. Wind speed, wave height, temperature and depth significantly contributed to the variation in larval fish densities. Abundances of larval fishes were greater offshore than onshore, larvae from pelagic eggs dominated catches offshore, while larvae from demersal eggs dominated onshore catches. Habitat structure strongly influenced the composition of larval fishes between the bays and abundances were significantly greater at night than during the day. Most larval fishes displayed a reverse diel vertical migration pattern and were most influenced by predators, wind speed and cloud cover. This study shows that larval fish assemblages are highly complex and patchy. Spawning mode, individual species behaviour, diel vertical migration, current structure, depth, temperature, wind speed, cloud cover and type of habitat substratum all influence larval fish composition and distribution in the nearshore waters of South Africa.
- Full Text:
- Date Issued: 2019
Analysis of bacterial Mur amide ligase enzymes for the identification of inhibitory compounds by in silico methods
- Chamboko, Chiratidzo Respina
- Authors: Chamboko, Chiratidzo Respina
- Date: 2020
- Subjects: Mur amide ligases , Ligases , Ligand binding (Biochemistry) , Antibacterial agents
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/163430 , vital:41036
- Description: An increased emergence of resistant pathogenic bacterial strains over the years has resulted in many people dying of untreatable infections. This has become one of the most critical global public health problems, as resistant strains are complicating treatment of infectious diseases, increasing human morbidity, mortality, and health care costs. A very limited amount of effective antibiotics is currently available, but the development of novel classes of antibacterial agents is becoming a priority. Mur amide ligases are enzymes that have been identified as potentially good targets for antibiotics, as they are uniquely found in bacteria. They are responsible for the formation of peptide bonds in a growing peptidoglycan structure for bacterial cell walls. The current work presented here focused on characterizing these Mur amide ligase enzymes and obtaining inhibitory compounds that could potentially be of use in drug discovery of antibacterial agents. To do this, multiple sequence alignment, motif analysis and phylogenetic tree constructions were carried out, followed by docking studies and molecular dynamic simulations. Prior to docking, homology modelling of missing residues in the MurF structure (PDB 1GG4) was performed. Characterization results revealed the Mur amide ligase enzymes contained defined conservation in limited regions, that ultimately mapped towards the central domain responsible for ATP binding (presence of a conserved GKT motif). Further analysis of results further unraveled the unique patterns observed within each group of the family of enzymes. As a result of these findings, docking studies were carried out on each Mur amide ligase structure. At most, two ligands were identified to be sufficiently inhibiting each Mur amide ligase. The ligands obtained were SANC00574 and SANC00575 for MurC, SANC00290 and SANC00438 for MurD, SANC00290 and SANC00525 for MurE and SANC00290 and SANC00434 for MurF. The two best ligands identified for each enzyme had docked in the active site of their respective proteins, passed Lipinski’s rule of five and had substantially low binding energies. Molecular dynamic simulations were then performed to analyze the behavior of the proteins and protein-ligand complexes, to confirm the lead compounds as good inhibitors of the Mur amide ligases. In the case of MurC, MurD and MurE complexes, the identified ligands clearly impacted the behavior of the protein, as the ligand bound proteins became more compact and stable, while flexibility decreased. There was however an opposite effect on MurF complexes, that resulted in identified inhibitors being discarded. As a potential next step, in vivo and in vitro experiments can be performed with identified ligands from this research, to further support the information presented.
- Full Text:
- Date Issued: 2020
- Authors: Chamboko, Chiratidzo Respina
- Date: 2020
- Subjects: Mur amide ligases , Ligases , Ligand binding (Biochemistry) , Antibacterial agents
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/163430 , vital:41036
- Description: An increased emergence of resistant pathogenic bacterial strains over the years has resulted in many people dying of untreatable infections. This has become one of the most critical global public health problems, as resistant strains are complicating treatment of infectious diseases, increasing human morbidity, mortality, and health care costs. A very limited amount of effective antibiotics is currently available, but the development of novel classes of antibacterial agents is becoming a priority. Mur amide ligases are enzymes that have been identified as potentially good targets for antibiotics, as they are uniquely found in bacteria. They are responsible for the formation of peptide bonds in a growing peptidoglycan structure for bacterial cell walls. The current work presented here focused on characterizing these Mur amide ligase enzymes and obtaining inhibitory compounds that could potentially be of use in drug discovery of antibacterial agents. To do this, multiple sequence alignment, motif analysis and phylogenetic tree constructions were carried out, followed by docking studies and molecular dynamic simulations. Prior to docking, homology modelling of missing residues in the MurF structure (PDB 1GG4) was performed. Characterization results revealed the Mur amide ligase enzymes contained defined conservation in limited regions, that ultimately mapped towards the central domain responsible for ATP binding (presence of a conserved GKT motif). Further analysis of results further unraveled the unique patterns observed within each group of the family of enzymes. As a result of these findings, docking studies were carried out on each Mur amide ligase structure. At most, two ligands were identified to be sufficiently inhibiting each Mur amide ligase. The ligands obtained were SANC00574 and SANC00575 for MurC, SANC00290 and SANC00438 for MurD, SANC00290 and SANC00525 for MurE and SANC00290 and SANC00434 for MurF. The two best ligands identified for each enzyme had docked in the active site of their respective proteins, passed Lipinski’s rule of five and had substantially low binding energies. Molecular dynamic simulations were then performed to analyze the behavior of the proteins and protein-ligand complexes, to confirm the lead compounds as good inhibitors of the Mur amide ligases. In the case of MurC, MurD and MurE complexes, the identified ligands clearly impacted the behavior of the protein, as the ligand bound proteins became more compact and stable, while flexibility decreased. There was however an opposite effect on MurF complexes, that resulted in identified inhibitors being discarded. As a potential next step, in vivo and in vitro experiments can be performed with identified ligands from this research, to further support the information presented.
- Full Text:
- Date Issued: 2020
Identification of possible natural compounds as potential inhibitors against Plasmodium M1 alanyl aminopeptidase
- Soliman, Omar Samir Abdel Ghaffar
- Authors: Soliman, Omar Samir Abdel Ghaffar
- Date: 2019
- Subjects: Plasmodium , Malaria -- Chemotherapy , Plasmodium -- Inhibitors , Drug resistance in microorganisms , Aminopeptidases
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/72284 , vital:30026
- Description: Malaria is a major tropical health problem with a 29% mortality rate among people of all ages; it also affects 35% of the children. Despite the decrease in mortality rate in recent years, malaria still results in around 2000 deaths per day. Malaria is caused by Plasmodium parasites and is transmitted to humans via the bites from infected female Anopheles mosquitoes during blood meals. There are five different Plasmodium species that can cause human malaria, which include Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale and Plasmodium knowlesi. Among these five species, the most pathogenic ones are Plasmodium falciparum and Plasmodium vivax. Malaria is usually hard to diagnose because the symptoms are not exclusive to malaria and very similar to flu, e.g., fever, muscle pain, and chills, which lead to the misdiagnosis of malaria cases. Malaria is lethal if not treated because it can cause severe complications in the respiratory tract, liver, metabolic acidosis, and hypoglycemia. The malaria parasite life cycle includes two types of hosts, i.e., a human host and female Anopheles mosquito host. Malaria continuously develops resistance to the available drugs, which is one of the major challenges in disease control. This situation confirms the need to develop new drugs that target virulence factors of malaria. The malarial parasite has three main life cycle stages, which include the host liver stage, host blood stage and vector stage. In the blood stage, parasites degrade hemoglobin to amino acids, which is important as these parasites cannot produce their own amino acids. Different proteases are involved in this hemoglobin degradation process. M1 alanyl aminopeptidase is one of these proteases involved at the end of hemoglobin degradation. This study focused on M1 alanyl aminopeptidase as a potential drug target. M1 alanyl aminopeptidase consists of four domains: N-terminal domain, catalytic domain, middle domain and C-terminal domain. The catalytic domain remains conserved among different Plasmodium species. Inhibition of this enzyme might prevent Plasmodium growth as it can’t produce its own amino acids. In this study, sequence analysis was carried out in both human and Plasmodium M1 alanyl aminopeptidase to identify conserved and divergent regions between them. 3D protein models of the M1 alanyl aminopeptidase from Plasmodium species were built and validated. Then the generated models were used for virtual screening against 623 compounds retrieved from the South African Natural Compounds Database (SANCDB, https://sancdb.rubi.ru.ac.za/). Virtual screening was done using blind and targeted docking methods. Docking was used to identify compounds with selective high binding affinity to the active site of the parasite protein. In this study, one SANCDB compound was selected for each protein: SANC00531 was selected against P. falciparum M1 alanyl aminopeptidase, SANC00469 against P. knowlesi, SANC00660 against P. vivax, SANC00144 against P. ovale and SANC00109 against P. malariae. It was found that Plamsodium M1 alanyl aminopeptidase can be used as a potential drug target as it showed selective binding against different inhibitor compounds. This result will be investigated in future work though molecular dynamic analysis to investigate the stability of protein-ligand complexes.
- Full Text:
- Date Issued: 2019
- Authors: Soliman, Omar Samir Abdel Ghaffar
- Date: 2019
- Subjects: Plasmodium , Malaria -- Chemotherapy , Plasmodium -- Inhibitors , Drug resistance in microorganisms , Aminopeptidases
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/72284 , vital:30026
- Description: Malaria is a major tropical health problem with a 29% mortality rate among people of all ages; it also affects 35% of the children. Despite the decrease in mortality rate in recent years, malaria still results in around 2000 deaths per day. Malaria is caused by Plasmodium parasites and is transmitted to humans via the bites from infected female Anopheles mosquitoes during blood meals. There are five different Plasmodium species that can cause human malaria, which include Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale and Plasmodium knowlesi. Among these five species, the most pathogenic ones are Plasmodium falciparum and Plasmodium vivax. Malaria is usually hard to diagnose because the symptoms are not exclusive to malaria and very similar to flu, e.g., fever, muscle pain, and chills, which lead to the misdiagnosis of malaria cases. Malaria is lethal if not treated because it can cause severe complications in the respiratory tract, liver, metabolic acidosis, and hypoglycemia. The malaria parasite life cycle includes two types of hosts, i.e., a human host and female Anopheles mosquito host. Malaria continuously develops resistance to the available drugs, which is one of the major challenges in disease control. This situation confirms the need to develop new drugs that target virulence factors of malaria. The malarial parasite has three main life cycle stages, which include the host liver stage, host blood stage and vector stage. In the blood stage, parasites degrade hemoglobin to amino acids, which is important as these parasites cannot produce their own amino acids. Different proteases are involved in this hemoglobin degradation process. M1 alanyl aminopeptidase is one of these proteases involved at the end of hemoglobin degradation. This study focused on M1 alanyl aminopeptidase as a potential drug target. M1 alanyl aminopeptidase consists of four domains: N-terminal domain, catalytic domain, middle domain and C-terminal domain. The catalytic domain remains conserved among different Plasmodium species. Inhibition of this enzyme might prevent Plasmodium growth as it can’t produce its own amino acids. In this study, sequence analysis was carried out in both human and Plasmodium M1 alanyl aminopeptidase to identify conserved and divergent regions between them. 3D protein models of the M1 alanyl aminopeptidase from Plasmodium species were built and validated. Then the generated models were used for virtual screening against 623 compounds retrieved from the South African Natural Compounds Database (SANCDB, https://sancdb.rubi.ru.ac.za/). Virtual screening was done using blind and targeted docking methods. Docking was used to identify compounds with selective high binding affinity to the active site of the parasite protein. In this study, one SANCDB compound was selected for each protein: SANC00531 was selected against P. falciparum M1 alanyl aminopeptidase, SANC00469 against P. knowlesi, SANC00660 against P. vivax, SANC00144 against P. ovale and SANC00109 against P. malariae. It was found that Plamsodium M1 alanyl aminopeptidase can be used as a potential drug target as it showed selective binding against different inhibitor compounds. This result will be investigated in future work though molecular dynamic analysis to investigate the stability of protein-ligand complexes.
- Full Text:
- Date Issued: 2019
An investigation into the fire regimes of the upper Tsitsa River catchment
- Authors: Snyman, Gareth
- Date: 2020
- Subjects: Prescribed burning -- Environmental aspects -- South Africa , Fire ecology -- South Africa , Tsitsa River catchment (South Africa) , Soil erosion -- South Africa , Grasslands -- Management -- South Africa
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/145256 , vital:38422
- Description: South African grasslands are rich in flora and fauna and are a dominant vegetation cover in many of the water catchment areas, while providing a multitude of services to local communities and the environment. Fires have been the primary tool used to manage grasslands for livestock production for many years. However, there is debate about how they impact and alter landscapes and there are two schools of thought throughout literature. One argument is that fires are detrimental to landscapes; altering hydrological processes by causing excess soil erosion and changing soil properties. The other opinion is that fires are beneficial to ecosystems; maintaining vegetation structure, preventing bush encroachment, cycling nutrients and allowing for new plant growth. This study focused on the effects of fire regimes on soil properties in order to better understand the role that fire plays in geomorphic processes in the upper Tsitsa River catchment. The catchment falls under two land management types- commercial and communal, with the latter being severely degraded. Fire regimes were determined using a combination of Landsat and MODIS remotely sensed data and testing was carried out on soils exposed to different Fire Return Intervals (FRI). It was found that soils that were exposed to a high fire frequency (1-2-year FRI) exhibited a significantly higher degree of water repellency (p< 0.001) and surface hardness (p< 0.001) than soils that were exposed to a low fire frequency (3-4 year FRI), which indicates increased erosion potential. However, whilst a higher fire frequency resulted in altered soil characteristics, it contributed to landscape degradation as suggested in the literature, and soils are able to return to their previous state in over a short period of time. Further investigation into factors affecting fire regimes found that geology influenced vegetation type, resulting in differences in biomass in the two land management areas. The commercially managed land supported a high biomass, which resulted in a more natural fire regime with frequent fires, whilst the communally managed land supported less biomass. Smaller fuel loads and increased landscape fragmentation through overgrazing and road networks resulted in an altered fire regime with less frequent fires. Although the communally managed land has an altered fire regime, this research suggests that fires are not influencing the severe degradation present in these areas, and this is rather a function of geology and mismanagement of land. Whilst fire and grazing management plans can be implemented to alter the modified fire regime in the communal areas back to its natural state, this would take a long time and an increase in fires would pose a threat to surrounding communities. Overall the fire regimes in the upper Tsitsa River catchment are being altered by human influence and land management type, and whilst fire frequency is negatively altering soil properties, these soils can return to their natural state. The severe land degradation present in the communally managed areas are not a function of fire frequency and rather of geology and land management.
- Full Text:
- Date Issued: 2020
- Authors: Snyman, Gareth
- Date: 2020
- Subjects: Prescribed burning -- Environmental aspects -- South Africa , Fire ecology -- South Africa , Tsitsa River catchment (South Africa) , Soil erosion -- South Africa , Grasslands -- Management -- South Africa
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/145256 , vital:38422
- Description: South African grasslands are rich in flora and fauna and are a dominant vegetation cover in many of the water catchment areas, while providing a multitude of services to local communities and the environment. Fires have been the primary tool used to manage grasslands for livestock production for many years. However, there is debate about how they impact and alter landscapes and there are two schools of thought throughout literature. One argument is that fires are detrimental to landscapes; altering hydrological processes by causing excess soil erosion and changing soil properties. The other opinion is that fires are beneficial to ecosystems; maintaining vegetation structure, preventing bush encroachment, cycling nutrients and allowing for new plant growth. This study focused on the effects of fire regimes on soil properties in order to better understand the role that fire plays in geomorphic processes in the upper Tsitsa River catchment. The catchment falls under two land management types- commercial and communal, with the latter being severely degraded. Fire regimes were determined using a combination of Landsat and MODIS remotely sensed data and testing was carried out on soils exposed to different Fire Return Intervals (FRI). It was found that soils that were exposed to a high fire frequency (1-2-year FRI) exhibited a significantly higher degree of water repellency (p< 0.001) and surface hardness (p< 0.001) than soils that were exposed to a low fire frequency (3-4 year FRI), which indicates increased erosion potential. However, whilst a higher fire frequency resulted in altered soil characteristics, it contributed to landscape degradation as suggested in the literature, and soils are able to return to their previous state in over a short period of time. Further investigation into factors affecting fire regimes found that geology influenced vegetation type, resulting in differences in biomass in the two land management areas. The commercially managed land supported a high biomass, which resulted in a more natural fire regime with frequent fires, whilst the communally managed land supported less biomass. Smaller fuel loads and increased landscape fragmentation through overgrazing and road networks resulted in an altered fire regime with less frequent fires. Although the communally managed land has an altered fire regime, this research suggests that fires are not influencing the severe degradation present in these areas, and this is rather a function of geology and mismanagement of land. Whilst fire and grazing management plans can be implemented to alter the modified fire regime in the communal areas back to its natural state, this would take a long time and an increase in fires would pose a threat to surrounding communities. Overall the fire regimes in the upper Tsitsa River catchment are being altered by human influence and land management type, and whilst fire frequency is negatively altering soil properties, these soils can return to their natural state. The severe land degradation present in the communally managed areas are not a function of fire frequency and rather of geology and land management.
- Full Text:
- Date Issued: 2020
Analysis of bacterial Mur amide ligase enzymes for the identification of inhibitory compounds by in silico methods
- Chamboko, Chiratidzo Respina
- Authors: Chamboko, Chiratidzo Respina
- Date: 2020
- Subjects: Pathogenic microorganisms -- Analysis , Drug resistance in microorganisms , Microorganisms -- Effect of drugs on , Antibiotics -- Effectiveness , Pathogenic bacteria , Drug tolerance , Enzymes -- Analysis , Peptide antibiotics
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/161911 , vital:40690
- Description: An increased emergence of resistant pathogenic bacterial strains over the years has resulted in many people dying of untreatable infections. This has become one of the most critical global public health problems, as resistant strains are complicating treatment of infectious diseases, increasing human morbidity, mortality, and health care costs. A very limited amount of effective antibiotics is currently available, but the development of novel classes of antibacterial agents is becoming a priority. Mur amide ligases are enzymes that have been identified as potentially good targets for antibiotics, as they are uniquely found in bacteria. They are responsible for the formation of peptide bonds in a growing peptidoglycan structure for bacterial cell walls. The current work presented here focused on characterizing these Mur amide ligase enzymes and obtaining inhibitory compounds that could potentially be of use in drug discovery of antibacterial agents. To do this, multiple sequence alignment, motif analysis and phylogenetic tree constructions were carried out, followed by docking studies and molecular dynamic simulations. Prior to docking, homology modelling of missing residues in the MurF structure (PDB 1GG4) was performed. Characterization results revealed the Mur amide ligase enzymes contained defined conservation in limited regions, that ultimately mapped towards the central domain responsible for ATP binding (presence of a conserved GKT motif). Further analysis of results further unraveled the unique patterns observed within each group of the family of enzymes. As a result of these findings, docking studies were carried out on each Mur amide ligase structure. At most, two ligands were identified to be sufficiently inhibiting each Mur amide ligase. The ligands obtained were SANC00574 and SANC00575 for MurC, SANC00290 and SANC00438 for MurD, SANC00290 and SANC00525 for MurE and SANC00290 and SANC00434 for MurF. The two best ligands identified for each enzyme had docked in the active site of their respective proteins, passed Lipinski’s rule of five and had substantially low binding energies. Molecular dynamic simulations were then performed to analyze the behavior of the proteins and protein-ligand complexes, to confirm the lead compounds as good inhibitors of the Mur amide ligases. In the case of MurC, MurD and MurE complexes, the identified ligands clearly impacted the behavior of the protein, as the ligand bound proteins became more compact and stable, while flexibility decreased. There was however an opposite effect on MurF complexes, that resulted in identified inhibitors being discarded. As a potential next step, in vivo and in vitro experiments can be performed with identified ligands from this research, to further support the information presented.
- Full Text:
- Date Issued: 2020
- Authors: Chamboko, Chiratidzo Respina
- Date: 2020
- Subjects: Pathogenic microorganisms -- Analysis , Drug resistance in microorganisms , Microorganisms -- Effect of drugs on , Antibiotics -- Effectiveness , Pathogenic bacteria , Drug tolerance , Enzymes -- Analysis , Peptide antibiotics
- Language: English
- Type: text , Thesis , Masters , MSc
- Identifier: http://hdl.handle.net/10962/161911 , vital:40690
- Description: An increased emergence of resistant pathogenic bacterial strains over the years has resulted in many people dying of untreatable infections. This has become one of the most critical global public health problems, as resistant strains are complicating treatment of infectious diseases, increasing human morbidity, mortality, and health care costs. A very limited amount of effective antibiotics is currently available, but the development of novel classes of antibacterial agents is becoming a priority. Mur amide ligases are enzymes that have been identified as potentially good targets for antibiotics, as they are uniquely found in bacteria. They are responsible for the formation of peptide bonds in a growing peptidoglycan structure for bacterial cell walls. The current work presented here focused on characterizing these Mur amide ligase enzymes and obtaining inhibitory compounds that could potentially be of use in drug discovery of antibacterial agents. To do this, multiple sequence alignment, motif analysis and phylogenetic tree constructions were carried out, followed by docking studies and molecular dynamic simulations. Prior to docking, homology modelling of missing residues in the MurF structure (PDB 1GG4) was performed. Characterization results revealed the Mur amide ligase enzymes contained defined conservation in limited regions, that ultimately mapped towards the central domain responsible for ATP binding (presence of a conserved GKT motif). Further analysis of results further unraveled the unique patterns observed within each group of the family of enzymes. As a result of these findings, docking studies were carried out on each Mur amide ligase structure. At most, two ligands were identified to be sufficiently inhibiting each Mur amide ligase. The ligands obtained were SANC00574 and SANC00575 for MurC, SANC00290 and SANC00438 for MurD, SANC00290 and SANC00525 for MurE and SANC00290 and SANC00434 for MurF. The two best ligands identified for each enzyme had docked in the active site of their respective proteins, passed Lipinski’s rule of five and had substantially low binding energies. Molecular dynamic simulations were then performed to analyze the behavior of the proteins and protein-ligand complexes, to confirm the lead compounds as good inhibitors of the Mur amide ligases. In the case of MurC, MurD and MurE complexes, the identified ligands clearly impacted the behavior of the protein, as the ligand bound proteins became more compact and stable, while flexibility decreased. There was however an opposite effect on MurF complexes, that resulted in identified inhibitors being discarded. As a potential next step, in vivo and in vitro experiments can be performed with identified ligands from this research, to further support the information presented.
- Full Text:
- Date Issued: 2020
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