Keratinous poultry wastes valorization through novel keratinases of Chryseobacterium cucumeris and Sphingobacterium multivorum isolated from poultry sludge
- Hendrick, Qaphela https://orcid.org/0000-0001-7529-8129
- Authors: Hendrick, Qaphela https://orcid.org/0000-0001-7529-8129
- Date: 2021-02
- Subjects: Agricultural wastes , Factory and trade waste
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/21400 , vital:48537
- Description: Annually, about 55percent of keratinous wastes are generated from various agro-industrial processing farms in South Africa. These wastes are difficult to handle due to structural integrity; hence, they constitute environmental issues due to the disposal means. Degradation of keratinous wastes using microbial-based technology has been deemed advantageous as it generates products with high-end values. Therefore, in this study, chicken feather and soil samples were collected from a local poultry farm, and bacteria were isolated using basal salt media supplemented with chicken feathers. The isolates were evaluated for proteolytic and keratinolytic potentials. The potent isolates were identified through 16S rDNA sequence and phylogenetic analysis. Fermentation media were optimized for enhanced keratinase production, and the amino acids liberated in the media during feather biodegradation were quantified. The biochemical properties of the keratinases produced were likewise determined. Ten (10) proteolytic bacteria were obtained from 20 isolates recovered from the samples with a diameter of halo on skimmed milk agar plate ranging from 15.5 ± 0.71 (mm) for isolate coded as PSW-15 to 28 ± 1.41 (mm) for isolate coded as PSW-08. The proteolytic bacteria showed variable keratinolytic potentials with percentage feather degradation that ranged from 29percent for PSW-11 to 84percent for PSW-14, and keratinase activity ranging from 99.99 U/mL for PSW-15 to 761.82 U/mL for PSW-14. The most potent isolates coded as PSW-14 and PSW-16 were identified as Chryseobacterium cucumeris FHN1 and Sphingobacterium multivorum HNFx. Their nucleotide sequences were submitted to the GenBank as MW16587 and MK82939, respectively. The optimization of fermentation conditions; C. cucumeris FHN1 showed improved activity at pH 5 - 6, inoculum size (4percent, v/v), chicken feather concentration (1percent, w/v), fermentation temperature (25o C). Similarly, S. multivorum HNFx showed optimal activity at pH 4.0, inoculum size (5percent, v/v), chicken feather concentration (2.5percent, w/v), and fermentation temperature (25-30 oC). C. cucumeris FHN1 and S. multivorum HNFx showed maximum keratinase production of 485.54 U/mL and 526.36 U/mL at 96 h and 72 h of incubation period respectively. Serine, aspartic acid, glutamic acid, and proline were the most abundant amino acids in the degraded chicken feathers, and upon quantitation, the following concentration was respectively obtained; 3.71, 3.4, 4.19 and 4.35 (g/100g sample) against C. cucumeris FHN1. While S. multivorum HNFx yielded aspartic acid (2.04 g/100g sample) and glutamic acid (2.0 g/100g sample) in high concentration. The keratinases showed optimal catalytic efficiency at pH and temperature of 8.0 and 90 oC, respectively. C. cucumeris FHN1 keratinase was inhibited by metal ion chelating agents; EDTA and 1,10-phenanthroline, suggesting a metallo-type of protease. The enzyme showed remarkable stability after pre-treatment with DTT, Fe2+, Fe3+ and Cu2+, with respective residual activity of 108percent, 102percent, 114percent, and 104percent. The S. multivorum HNFx keratinase; activity was not inhibited by serine- and metallo-protease inhibitors. It maintained the following residual activity against the following chemical agents; DTT (124percent), hydrogen peroxide (152percent), DMSO (161percent), triton X-100 (152percent), tween-80 (101percent), and metal ions; Fe2+ (128percent), Fe3+ (104percent), K+ (117percent), Ca2+ (104percent), Na+ (103percent), Ba2+ (115percent), Al3+ (126percent). The enzyme showed a substantial loss of catalytic efficiency after pre-incubation with various laundry detergents. The keratinases' remarkable stability in the presence of various chemical agents and metal ions tested suggests biotechnological and industrial application potentials. Consequently, the isolates portend industrial relevance for keratinous waste valorization and an excellent source of keratinases of industrial relevance. , Thesis (MSc) -- Faculty of Science and Agriculture, 2021
- Full Text:
- Date Issued: 2021-02
- Authors: Hendrick, Qaphela https://orcid.org/0000-0001-7529-8129
- Date: 2021-02
- Subjects: Agricultural wastes , Factory and trade waste
- Language: English
- Type: Master's theses , text
- Identifier: http://hdl.handle.net/10353/21400 , vital:48537
- Description: Annually, about 55percent of keratinous wastes are generated from various agro-industrial processing farms in South Africa. These wastes are difficult to handle due to structural integrity; hence, they constitute environmental issues due to the disposal means. Degradation of keratinous wastes using microbial-based technology has been deemed advantageous as it generates products with high-end values. Therefore, in this study, chicken feather and soil samples were collected from a local poultry farm, and bacteria were isolated using basal salt media supplemented with chicken feathers. The isolates were evaluated for proteolytic and keratinolytic potentials. The potent isolates were identified through 16S rDNA sequence and phylogenetic analysis. Fermentation media were optimized for enhanced keratinase production, and the amino acids liberated in the media during feather biodegradation were quantified. The biochemical properties of the keratinases produced were likewise determined. Ten (10) proteolytic bacteria were obtained from 20 isolates recovered from the samples with a diameter of halo on skimmed milk agar plate ranging from 15.5 ± 0.71 (mm) for isolate coded as PSW-15 to 28 ± 1.41 (mm) for isolate coded as PSW-08. The proteolytic bacteria showed variable keratinolytic potentials with percentage feather degradation that ranged from 29percent for PSW-11 to 84percent for PSW-14, and keratinase activity ranging from 99.99 U/mL for PSW-15 to 761.82 U/mL for PSW-14. The most potent isolates coded as PSW-14 and PSW-16 were identified as Chryseobacterium cucumeris FHN1 and Sphingobacterium multivorum HNFx. Their nucleotide sequences were submitted to the GenBank as MW16587 and MK82939, respectively. The optimization of fermentation conditions; C. cucumeris FHN1 showed improved activity at pH 5 - 6, inoculum size (4percent, v/v), chicken feather concentration (1percent, w/v), fermentation temperature (25o C). Similarly, S. multivorum HNFx showed optimal activity at pH 4.0, inoculum size (5percent, v/v), chicken feather concentration (2.5percent, w/v), and fermentation temperature (25-30 oC). C. cucumeris FHN1 and S. multivorum HNFx showed maximum keratinase production of 485.54 U/mL and 526.36 U/mL at 96 h and 72 h of incubation period respectively. Serine, aspartic acid, glutamic acid, and proline were the most abundant amino acids in the degraded chicken feathers, and upon quantitation, the following concentration was respectively obtained; 3.71, 3.4, 4.19 and 4.35 (g/100g sample) against C. cucumeris FHN1. While S. multivorum HNFx yielded aspartic acid (2.04 g/100g sample) and glutamic acid (2.0 g/100g sample) in high concentration. The keratinases showed optimal catalytic efficiency at pH and temperature of 8.0 and 90 oC, respectively. C. cucumeris FHN1 keratinase was inhibited by metal ion chelating agents; EDTA and 1,10-phenanthroline, suggesting a metallo-type of protease. The enzyme showed remarkable stability after pre-treatment with DTT, Fe2+, Fe3+ and Cu2+, with respective residual activity of 108percent, 102percent, 114percent, and 104percent. The S. multivorum HNFx keratinase; activity was not inhibited by serine- and metallo-protease inhibitors. It maintained the following residual activity against the following chemical agents; DTT (124percent), hydrogen peroxide (152percent), DMSO (161percent), triton X-100 (152percent), tween-80 (101percent), and metal ions; Fe2+ (128percent), Fe3+ (104percent), K+ (117percent), Ca2+ (104percent), Na+ (103percent), Ba2+ (115percent), Al3+ (126percent). The enzyme showed a substantial loss of catalytic efficiency after pre-incubation with various laundry detergents. The keratinases' remarkable stability in the presence of various chemical agents and metal ions tested suggests biotechnological and industrial application potentials. Consequently, the isolates portend industrial relevance for keratinous waste valorization and an excellent source of keratinases of industrial relevance. , Thesis (MSc) -- Faculty of Science and Agriculture, 2021
- Full Text:
- Date Issued: 2021-02
Community-driven solid waste management initiatives in Missionvale
- Authors: Kwava, Eunice
- Date: 2020
- Subjects: Factory and trade waste
- Language: English
- Type: Thesis , Masters , MDS
- Identifier: http://hdl.handle.net/10948/48709 , vital:41064
- Description: Solid waste management is particularly challenging in poor communities where socio-economic issues take precedence over keeping a clean environment. This study explores the effectiveness of community-driven waste management strategies that are implemented in Missionvale, a poverty-stricken township that forms part of the Nelson Mandela Metropole, located in South Africa’s Eastern Cape Province. The aim of this qualitative study was to explore the sustainability of the community-driven waste management practices in the township. This was premised on the understanding that having the community at the centre of waste management is important and community initiatives coupled with support from key stakeholders, yields the desired outcome: sustainable waste management. The objectives of the study were: (i) to understand what happens to the solid waste generated in Missionvale; (ii) to assess the effects of the waste and its disposal (or lack thereof) in Missionvale; (iii) to identify the challenges faced by the public in implementing waste management initiatives for sustainable development; and (iv) to establish the waste management practices in Missionvale and their effectiveness. The study found that whilst waste management is a secondary, less important issue to community respondents who are struggling with more pressing socio-economic hurdles, there are nevertheless efforts from informal waste pickers, local businesses, non-governmental entities and some individual community members to keep their environment clean. This occurs against the backdrop of the municipality’s failure in its task to provide consistent waste collection and disposal.
- Full Text:
- Date Issued: 2020
- Authors: Kwava, Eunice
- Date: 2020
- Subjects: Factory and trade waste
- Language: English
- Type: Thesis , Masters , MDS
- Identifier: http://hdl.handle.net/10948/48709 , vital:41064
- Description: Solid waste management is particularly challenging in poor communities where socio-economic issues take precedence over keeping a clean environment. This study explores the effectiveness of community-driven waste management strategies that are implemented in Missionvale, a poverty-stricken township that forms part of the Nelson Mandela Metropole, located in South Africa’s Eastern Cape Province. The aim of this qualitative study was to explore the sustainability of the community-driven waste management practices in the township. This was premised on the understanding that having the community at the centre of waste management is important and community initiatives coupled with support from key stakeholders, yields the desired outcome: sustainable waste management. The objectives of the study were: (i) to understand what happens to the solid waste generated in Missionvale; (ii) to assess the effects of the waste and its disposal (or lack thereof) in Missionvale; (iii) to identify the challenges faced by the public in implementing waste management initiatives for sustainable development; and (iv) to establish the waste management practices in Missionvale and their effectiveness. The study found that whilst waste management is a secondary, less important issue to community respondents who are struggling with more pressing socio-economic hurdles, there are nevertheless efforts from informal waste pickers, local businesses, non-governmental entities and some individual community members to keep their environment clean. This occurs against the backdrop of the municipality’s failure in its task to provide consistent waste collection and disposal.
- Full Text:
- Date Issued: 2020
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