A polarimetric method for collagenase activity measurement
- Brüning, Adrian Rudolf Nicolaus Ernst
- Authors: Brüning, Adrian Rudolf Nicolaus Ernst
- Date: 1992
- Subjects: Collagenases -- Research , Hides and skins -- Preservation -- Research
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4052 , http://hdl.handle.net/10962/d1004113 , Collagenases -- Research , Hides and skins -- Preservation -- Research
- Description: A polarimetric method for monitoring the rate of soluble collagen breakdown by collagenase enzyme action has been developed. The method represents an extension of previous physicochemical techniques based on viscometry, but is simpler and easier to carry out, particularly in the case of reaction rate studies. The method was developed arising from reports of collagenase activity measurement on inappropriate substrates such as gelatin, modified collagens and synthetic polypeptides. The optical method depends on measurement of the loss in optical rotation in solutions of soluble calfskin collagen resulting from initial enzymic cleavage of the collagen trip1e-helix, followed by spontaneous unwinding of the resultant unstable helical fragments. Specific assay conditions were chosen to ensure that the loss in optical rotation following enzymic cleavage was rapid and complete. The method is specific since in the absence of collagenase, non-specific proteinases produce only a limited decrease in solution optical activity. The method has also been compared with established physicochemical assay techniques and compares favourably with both viscometric and titrimetric collagenase assays. The availability of a rapid, sensitive and quantitative procedure for measurement of collagenase activity provides a convenient means for detecting the presence of collagenase in solution and examination of hide bacterial cultures for collagenase production. In addition, a study of biocidal compounds of potential interest in hide preservation for possible inhibitory effects on collagenase is conveniently carried out with the method. Fundamental research into synergistic action in enzymic hydrolysis of collagen is now possible, providing valuable insight into the mechanism of raw hide biodeterioration.
- Full Text:
- Date Issued: 1992
- Authors: Brüning, Adrian Rudolf Nicolaus Ernst
- Date: 1992
- Subjects: Collagenases -- Research , Hides and skins -- Preservation -- Research
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4052 , http://hdl.handle.net/10962/d1004113 , Collagenases -- Research , Hides and skins -- Preservation -- Research
- Description: A polarimetric method for monitoring the rate of soluble collagen breakdown by collagenase enzyme action has been developed. The method represents an extension of previous physicochemical techniques based on viscometry, but is simpler and easier to carry out, particularly in the case of reaction rate studies. The method was developed arising from reports of collagenase activity measurement on inappropriate substrates such as gelatin, modified collagens and synthetic polypeptides. The optical method depends on measurement of the loss in optical rotation in solutions of soluble calfskin collagen resulting from initial enzymic cleavage of the collagen trip1e-helix, followed by spontaneous unwinding of the resultant unstable helical fragments. Specific assay conditions were chosen to ensure that the loss in optical rotation following enzymic cleavage was rapid and complete. The method is specific since in the absence of collagenase, non-specific proteinases produce only a limited decrease in solution optical activity. The method has also been compared with established physicochemical assay techniques and compares favourably with both viscometric and titrimetric collagenase assays. The availability of a rapid, sensitive and quantitative procedure for measurement of collagenase activity provides a convenient means for detecting the presence of collagenase in solution and examination of hide bacterial cultures for collagenase production. In addition, a study of biocidal compounds of potential interest in hide preservation for possible inhibitory effects on collagenase is conveniently carried out with the method. Fundamental research into synergistic action in enzymic hydrolysis of collagen is now possible, providing valuable insight into the mechanism of raw hide biodeterioration.
- Full Text:
- Date Issued: 1992
Regulation of the indoleamines by sex steroids
- Authors: Awah, Edmund Kpabi
- Date: 1992
- Subjects: Steroids -- Research , Steroid drugs -- Research
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4053 , http://hdl.handle.net/10962/d1004114 , Steroids -- Research , Steroid drugs -- Research
- Description: Alteration of serum tryptophan leads to parallel alterations in brain tryptophan levels. Such changes in brain tryptophan levels has been shown to lead to mood disturbances. The primary enzyme responsible for altering serum tryptophan levels is the liver cytosolic enzyme, tryptophan pyrrolase. Activation of this enzyme is responsible for the enhanced catabolism of circulating tryptophan. The purpose of the present study was firstly to establish whether there is a link between sex steroids and tryptophan pyrrolase activity especially since sex steroids are also known to cause mood disturbances and secondly to determine the effects of sex steroids on brain indolamine metabolism. The results show that all three sex steroids induce the activity of tryptophan pyrrolase implying that they decrease serum tryptophan levels by the activation of tryptophan pyrrolase, thus making less tryptophan available for uptake by the brain. It was also shown that the sex steroids enhance the uptake of ¹⁴C-tryptophan by brain synatopsomes. In addition, the sex steroids influenced the pattern of metabolism of serotonin by organ cultures of rat pineal glands. It is possible that the sex steroids regulate the availability and uptake of indoleamines in the brain.
- Full Text:
- Date Issued: 1992
- Authors: Awah, Edmund Kpabi
- Date: 1992
- Subjects: Steroids -- Research , Steroid drugs -- Research
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4053 , http://hdl.handle.net/10962/d1004114 , Steroids -- Research , Steroid drugs -- Research
- Description: Alteration of serum tryptophan leads to parallel alterations in brain tryptophan levels. Such changes in brain tryptophan levels has been shown to lead to mood disturbances. The primary enzyme responsible for altering serum tryptophan levels is the liver cytosolic enzyme, tryptophan pyrrolase. Activation of this enzyme is responsible for the enhanced catabolism of circulating tryptophan. The purpose of the present study was firstly to establish whether there is a link between sex steroids and tryptophan pyrrolase activity especially since sex steroids are also known to cause mood disturbances and secondly to determine the effects of sex steroids on brain indolamine metabolism. The results show that all three sex steroids induce the activity of tryptophan pyrrolase implying that they decrease serum tryptophan levels by the activation of tryptophan pyrrolase, thus making less tryptophan available for uptake by the brain. It was also shown that the sex steroids enhance the uptake of ¹⁴C-tryptophan by brain synatopsomes. In addition, the sex steroids influenced the pattern of metabolism of serotonin by organ cultures of rat pineal glands. It is possible that the sex steroids regulate the availability and uptake of indoleamines in the brain.
- Full Text:
- Date Issued: 1992
The culture of Dunaliella salina and the production of β-carotene in tannery effluents
- Authors: Laubscher, Richard Keith
- Date: 1992
- Subjects: Dunaliella , Carotenes , Tanneries -- Waste disposal , Recycling (Waste, etc.)
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4055 , http://hdl.handle.net/10962/d1004116 , Dunaliella , Carotenes , Tanneries -- Waste disposal , Recycling (Waste, etc.)
- Description: The problems of waste disposal in the tanning industry are unique in that the effluents are highly saline, have a high organic loading and contain heavy metals. Methods are available for the safe treatment and disposal of the latter two components, but the saline component requires the expensive outlay of evaporation ponds. This study has identified a possible use for the saline effluents, turning a problematic waste product into a potentially valuable by-product. A range of tannery effluents were identified and tested for their suitability for the mass cultivation of Dunaliella salina (bardawil strain). The bardawil strain was preferred over a local isolate because of its higher production of β-carotene. Ponded tannery effluents and combined processes effluent proved unsuitable for realistic propagation of the alga. Anaerobic digestion of combined processes effluent did not improve its suitability significantly. Anaerobic digestion of hide-soak effluent may remove persistent antimicrobial agents which influence algal growth, but its contribution to enhancing algal growth is equivocal. Undigested hide-soak effluent lacking in persistent antimicrobial agents was found to be an ideal culture medium, as no additional nutrients needed to be added. Significantly higher biomass was obtained in this effluent compared to chemically defined media. Induction of β-carotene was achieved in nitrogen-deficient defined media after culture in tannery effluent. This suggests that a two-stage system using hide-soak effluent for cell propagation and nitrogen deficient media for β-carotene induction, could be possible for the mass cultivation of D. salina for β-carotene production.
- Full Text:
- Date Issued: 1992
- Authors: Laubscher, Richard Keith
- Date: 1992
- Subjects: Dunaliella , Carotenes , Tanneries -- Waste disposal , Recycling (Waste, etc.)
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4055 , http://hdl.handle.net/10962/d1004116 , Dunaliella , Carotenes , Tanneries -- Waste disposal , Recycling (Waste, etc.)
- Description: The problems of waste disposal in the tanning industry are unique in that the effluents are highly saline, have a high organic loading and contain heavy metals. Methods are available for the safe treatment and disposal of the latter two components, but the saline component requires the expensive outlay of evaporation ponds. This study has identified a possible use for the saline effluents, turning a problematic waste product into a potentially valuable by-product. A range of tannery effluents were identified and tested for their suitability for the mass cultivation of Dunaliella salina (bardawil strain). The bardawil strain was preferred over a local isolate because of its higher production of β-carotene. Ponded tannery effluents and combined processes effluent proved unsuitable for realistic propagation of the alga. Anaerobic digestion of combined processes effluent did not improve its suitability significantly. Anaerobic digestion of hide-soak effluent may remove persistent antimicrobial agents which influence algal growth, but its contribution to enhancing algal growth is equivocal. Undigested hide-soak effluent lacking in persistent antimicrobial agents was found to be an ideal culture medium, as no additional nutrients needed to be added. Significantly higher biomass was obtained in this effluent compared to chemically defined media. Induction of β-carotene was achieved in nitrogen-deficient defined media after culture in tannery effluent. This suggests that a two-stage system using hide-soak effluent for cell propagation and nitrogen deficient media for β-carotene induction, could be possible for the mass cultivation of D. salina for β-carotene production.
- Full Text:
- Date Issued: 1992
The microbial production of polyphenol oxidase enzyme systems and their application in the treatment of phenolic wastewaters
- Scherman, Patricia Ann (neé Goetch)
- Authors: Scherman, Patricia Ann (neé Goetch)
- Date: 1992
- Subjects: Phenol oxidase Water -- Purification -- Biological treatment Enzymes -- Regulation
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4047 , http://hdl.handle.net/10962/d1004108
- Description: Phenolic compounds are a group of organic chemicals present in the wastewaters of many synthetic industrial processes. Due to their extreme toxicity to man and animals, and deleterious impact on the environment, a range of techniques exist for the effective treatment and disposal of these pollutants. Biological degradation using microbial enzymes presents a valuable alternative to conventional wastewater treatment systems. This research was therefore initiated to investigate the polyphenol oxidase enzyme system and the feasibility of its application for effluent treatment and studies in organic solvents. The enzyme system is widely distributed in nature, with Agaricus bisporus (the common mushroom) being the best known producer. Biochemical investigations of the enzyme system were therefore carried out using this extract. A screening programme was initiated to identify microbial polyphenol oxidase producers which could be cultured in liquid media, thereby enabling the production of large quantities of enzyme in fermentation systems. Extensive growth optimization and enzyme induction and optimization studies were carried out on selected cultures. A number of good producers were isolated, namely a bacterial culture designated AECI culture no. 26, Streptomyces antibioticus, Streptomyces glaucescens and a manipulated strain, Streptomyces lividans (pIJ702). Enzyme production by Agaricus bisporus mycelia was optimized in deep-liquid culture; enzyme extracts showed high phenol removal efficiencies. Streptomyces antibioticus, Streptomyces glaucescens, Streptomyces lividans (pIJ702) and AECI culture no. 26 whole cells were also investigated for phenol-removing ability in simulated phenolic effluents. The use of whole cells reduces enzyme inactivation and instability due to the protection of the enzyme system within the cell. All cultures showed improved removal efficiencies in phenolic growth media. These results strongly suggest their use for phenol removal in continuous systems.
- Full Text:
- Date Issued: 1992
- Authors: Scherman, Patricia Ann (neé Goetch)
- Date: 1992
- Subjects: Phenol oxidase Water -- Purification -- Biological treatment Enzymes -- Regulation
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4047 , http://hdl.handle.net/10962/d1004108
- Description: Phenolic compounds are a group of organic chemicals present in the wastewaters of many synthetic industrial processes. Due to their extreme toxicity to man and animals, and deleterious impact on the environment, a range of techniques exist for the effective treatment and disposal of these pollutants. Biological degradation using microbial enzymes presents a valuable alternative to conventional wastewater treatment systems. This research was therefore initiated to investigate the polyphenol oxidase enzyme system and the feasibility of its application for effluent treatment and studies in organic solvents. The enzyme system is widely distributed in nature, with Agaricus bisporus (the common mushroom) being the best known producer. Biochemical investigations of the enzyme system were therefore carried out using this extract. A screening programme was initiated to identify microbial polyphenol oxidase producers which could be cultured in liquid media, thereby enabling the production of large quantities of enzyme in fermentation systems. Extensive growth optimization and enzyme induction and optimization studies were carried out on selected cultures. A number of good producers were isolated, namely a bacterial culture designated AECI culture no. 26, Streptomyces antibioticus, Streptomyces glaucescens and a manipulated strain, Streptomyces lividans (pIJ702). Enzyme production by Agaricus bisporus mycelia was optimized in deep-liquid culture; enzyme extracts showed high phenol removal efficiencies. Streptomyces antibioticus, Streptomyces glaucescens, Streptomyces lividans (pIJ702) and AECI culture no. 26 whole cells were also investigated for phenol-removing ability in simulated phenolic effluents. The use of whole cells reduces enzyme inactivation and instability due to the protection of the enzyme system within the cell. All cultures showed improved removal efficiencies in phenolic growth media. These results strongly suggest their use for phenol removal in continuous systems.
- Full Text:
- Date Issued: 1992
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