Investigation of brewery waste grains and microbial fuel cells as value-additive technologies improving solvent production yields in Clostridium acetobutylicum (ATCC 824) fermentation
- Authors: Du Toit, Ryan Guillaume
- Date: 2023-10-13
- Subjects: Biomass energy , Butanol , Fermentation , Microbial fuel cells , Brewery waste , Clostridium acetobutylicum
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424643 , vital:72171
- Description: The production of the solvent compounds acetone, ethanol and butanol through fermentation of organic feedstocks using Clostridia species could be a promising route for biofuel production. However, the cost of raw materials, low yields and the complexity of anaerobic fermentation continue to hinder this means of generating these compounds. The research presented in this Thesis investigated low-cost interventions that could decrease the costs of production and to direct the synthesis of fuel compounds using microbial fuel cells. Low-cost anaerobic chambers were designed and constructed for the propagation and manipulation of Clostridium acetobutylicum, selected as a low-risk microbial catalyst. Fermentation was monitored using in situ pH measurements and a combination of turbidity measurements, nutrient assays (especially total carbohydrates) and HPLC-RI detection as a means of monitoring the consumption of nutrients (glucose), production of precursor compounds (butyric acid) and the formation of solvent molecules (acetone/ethanol and butanol) during fermentation by this organism. Brewer’s spent grains were tested as a sustainable and low-cost feedstock for solvent production, comparing the effects of sterilising before fermentation, or allowing resident microflora to remain during Clostridium-catalysed solvent production. Sterilised spent grains significantly improved the production of solvent molecules (e.g. 12.97 ± 0.38 g/L of butanol yielded, compared to 0.40 ± 0.33 g/L for defined media sampled during the solventogenic phase); compared to these, the use of non-sterilised brewer’s grain decreased both the reproducibility and yields of fermentation (8.66 ± 1.6 g/L of butanol). Microbial fuel cells were studied as a possible means of altering electron transfer to/from electrode-attached Clostridia to control the metabolic shift in bacteria from acidogenesis to solventogenesis. The base line MFC (11.00 ± 4.69 g/L) fermentation experiment did produce higher acetone/ethanol than the baseline batch experiment MB (5.47 ± 4.48 g/L), indicating an improvement to solvent production in C. acetobutylicum (ATCC 824) in a MFC fermentation. In this study, MFC-1 demonstrated remarkable superiority over MB in terms of butyric acid production, yielding significantly higher concentrations while also improving acetone and ethanol production. However, the enhanced butyric acid production did not correspond to significantly increased butanol yields when compared to batch fermentation of chemically defined media. These findings highlight the potential of MFC-1 as an efficient approach for enhancing the fermentative production of valuable compounds, with a particular focus on butyric acid and acetone/ethanol. , Thesis (MSc) -- Faculty of Science, Biotechnology Innovation Centre, 2023
- Full Text:
- Date Issued: 2023-10-13
- Authors: Du Toit, Ryan Guillaume
- Date: 2023-10-13
- Subjects: Biomass energy , Butanol , Fermentation , Microbial fuel cells , Brewery waste , Clostridium acetobutylicum
- Language: English
- Type: Academic theses , Master's theses , text
- Identifier: http://hdl.handle.net/10962/424643 , vital:72171
- Description: The production of the solvent compounds acetone, ethanol and butanol through fermentation of organic feedstocks using Clostridia species could be a promising route for biofuel production. However, the cost of raw materials, low yields and the complexity of anaerobic fermentation continue to hinder this means of generating these compounds. The research presented in this Thesis investigated low-cost interventions that could decrease the costs of production and to direct the synthesis of fuel compounds using microbial fuel cells. Low-cost anaerobic chambers were designed and constructed for the propagation and manipulation of Clostridium acetobutylicum, selected as a low-risk microbial catalyst. Fermentation was monitored using in situ pH measurements and a combination of turbidity measurements, nutrient assays (especially total carbohydrates) and HPLC-RI detection as a means of monitoring the consumption of nutrients (glucose), production of precursor compounds (butyric acid) and the formation of solvent molecules (acetone/ethanol and butanol) during fermentation by this organism. Brewer’s spent grains were tested as a sustainable and low-cost feedstock for solvent production, comparing the effects of sterilising before fermentation, or allowing resident microflora to remain during Clostridium-catalysed solvent production. Sterilised spent grains significantly improved the production of solvent molecules (e.g. 12.97 ± 0.38 g/L of butanol yielded, compared to 0.40 ± 0.33 g/L for defined media sampled during the solventogenic phase); compared to these, the use of non-sterilised brewer’s grain decreased both the reproducibility and yields of fermentation (8.66 ± 1.6 g/L of butanol). Microbial fuel cells were studied as a possible means of altering electron transfer to/from electrode-attached Clostridia to control the metabolic shift in bacteria from acidogenesis to solventogenesis. The base line MFC (11.00 ± 4.69 g/L) fermentation experiment did produce higher acetone/ethanol than the baseline batch experiment MB (5.47 ± 4.48 g/L), indicating an improvement to solvent production in C. acetobutylicum (ATCC 824) in a MFC fermentation. In this study, MFC-1 demonstrated remarkable superiority over MB in terms of butyric acid production, yielding significantly higher concentrations while also improving acetone and ethanol production. However, the enhanced butyric acid production did not correspond to significantly increased butanol yields when compared to batch fermentation of chemically defined media. These findings highlight the potential of MFC-1 as an efficient approach for enhancing the fermentative production of valuable compounds, with a particular focus on butyric acid and acetone/ethanol. , Thesis (MSc) -- Faculty of Science, Biotechnology Innovation Centre, 2023
- Full Text:
- Date Issued: 2023-10-13
Development of a process for the preparation of linalool from CIS-2-pinanol
- Authors: Buddoo, Subash Ramnarain
- Date: 2009
- Subjects: Odors , Perfumes -- History , Perfumes -- Formulae , Fermentation
- Language: English
- Type: Thesis , Doctoral , DTech
- Identifier: vital:10425 , http://hdl.handle.net/10948/d1016219
- Description: Linalool is a key intermediate for the production of important fragrance chemicals such as geraniol, nerol, geranial, and neral. Linalool can be produced via a two-step process from α-pinene which is a major component of crude sulphated turpentine (CST) a foul-smelling, volatile waste product of the pulp and paper industry. The key step in this process is the pyrolysis step which involves the isomerisation of cis-2-pinanol to linalool and requires high temperatures (600-650°C) and is not very selective due to the decomposition of the product itself under these conditions. A client of the CSIR, Teubes Pty. Ltd., is a manufacturer of flavour and fragrance compounds for the local and international fragrance market and expressed an interest in producing linalool since the company would then gain access to other valuable fragrance chemicals via relatively simple processes. Earlier work conducted by AECI, R & D did not meet with much success since the selectivity to linalool was very poor and the process could hardly be deemed as scalable. The main objective of this project was therefore to develop a process for the selective isomerisation of cis-2-pinanol to linalool with minimum by-product formation and using process equipment that could be scaled to full-scale production. Since cis-2- pinanol could not be purchased in sufficient quantities for process development, a process had to be developed for the bench-scale preparation of kilogram quantities of cis-2-pinanol from α-pinene obtained from the client. Although this synthesis formed a minor part of this investigation, several process improvements and innovations were introduced to produce high quality cis-2-pinanol, in very good yields at kilogram scale. A major part of this investigation was the design and set up of a pyrolyis rig capable of operating at elevated temperatures (400 - 750°C) for the evaluation of various process parameters. Various vaporizer, reactor, and condensation systems were evaluated for their ability to cope with the demanding conditions on a consistent basis. The initial part of the investigation was a screening exercise to evaluate various process parameters as well as solvents, materials of construction, catalysts, etc. A comprehensive statistical design was also conducted to determine the critical process parameters and the model obtained was used to predict the optimum conditions required for the preparation of in-specification product on a consistent basis. These conditions were used in the preparation of a 1kg sample which was required by theclient for market evaluation purposes. The use of a novel microreactor system was also evaluated for the pinanol pyrolysis reaction. To our knowledge, this is the first time that a microreactor has been successfully used for this type of reaction in the Fragrance industry and a patent application is being filed by the CSIR. The kinetics of the reaction in both the tubular reactor system and the microreactor system was investigated. Computer modelling studies on both the systems were also conducted. The raw material cost to produce a kilogram of linalool is $1.40. There is a significant margin of 60.8 percent between the raw material cost of linalool and the current selling price ($3.57/kg). This clearly indicates that the project is potentially feasible from an economic point of view and we can now proceed with confidence to the next stage which is the engineering design, building and commissioning of the large scale pyrolysis rig. The rest of the process steps will be conducted on existing equipment currently present at the CSIR’s large scale facility (Imbiza in Isando, Gauteng).
- Full Text:
- Date Issued: 2009
- Authors: Buddoo, Subash Ramnarain
- Date: 2009
- Subjects: Odors , Perfumes -- History , Perfumes -- Formulae , Fermentation
- Language: English
- Type: Thesis , Doctoral , DTech
- Identifier: vital:10425 , http://hdl.handle.net/10948/d1016219
- Description: Linalool is a key intermediate for the production of important fragrance chemicals such as geraniol, nerol, geranial, and neral. Linalool can be produced via a two-step process from α-pinene which is a major component of crude sulphated turpentine (CST) a foul-smelling, volatile waste product of the pulp and paper industry. The key step in this process is the pyrolysis step which involves the isomerisation of cis-2-pinanol to linalool and requires high temperatures (600-650°C) and is not very selective due to the decomposition of the product itself under these conditions. A client of the CSIR, Teubes Pty. Ltd., is a manufacturer of flavour and fragrance compounds for the local and international fragrance market and expressed an interest in producing linalool since the company would then gain access to other valuable fragrance chemicals via relatively simple processes. Earlier work conducted by AECI, R & D did not meet with much success since the selectivity to linalool was very poor and the process could hardly be deemed as scalable. The main objective of this project was therefore to develop a process for the selective isomerisation of cis-2-pinanol to linalool with minimum by-product formation and using process equipment that could be scaled to full-scale production. Since cis-2- pinanol could not be purchased in sufficient quantities for process development, a process had to be developed for the bench-scale preparation of kilogram quantities of cis-2-pinanol from α-pinene obtained from the client. Although this synthesis formed a minor part of this investigation, several process improvements and innovations were introduced to produce high quality cis-2-pinanol, in very good yields at kilogram scale. A major part of this investigation was the design and set up of a pyrolyis rig capable of operating at elevated temperatures (400 - 750°C) for the evaluation of various process parameters. Various vaporizer, reactor, and condensation systems were evaluated for their ability to cope with the demanding conditions on a consistent basis. The initial part of the investigation was a screening exercise to evaluate various process parameters as well as solvents, materials of construction, catalysts, etc. A comprehensive statistical design was also conducted to determine the critical process parameters and the model obtained was used to predict the optimum conditions required for the preparation of in-specification product on a consistent basis. These conditions were used in the preparation of a 1kg sample which was required by theclient for market evaluation purposes. The use of a novel microreactor system was also evaluated for the pinanol pyrolysis reaction. To our knowledge, this is the first time that a microreactor has been successfully used for this type of reaction in the Fragrance industry and a patent application is being filed by the CSIR. The kinetics of the reaction in both the tubular reactor system and the microreactor system was investigated. Computer modelling studies on both the systems were also conducted. The raw material cost to produce a kilogram of linalool is $1.40. There is a significant margin of 60.8 percent between the raw material cost of linalool and the current selling price ($3.57/kg). This clearly indicates that the project is potentially feasible from an economic point of view and we can now proceed with confidence to the next stage which is the engineering design, building and commissioning of the large scale pyrolysis rig. The rest of the process steps will be conducted on existing equipment currently present at the CSIR’s large scale facility (Imbiza in Isando, Gauteng).
- Full Text:
- Date Issued: 2009
The effect of hydrostatic carbon dioxide pressure and extracellular ethanol on the performance of the yeast strain Saccharomyces cerevisiae during fermentation
- Authors: Longden, Nicholas Guy
- Date: 1993
- Subjects: Brewing -- Microbiology , Yeast , Fermentation , Saccharomyces cerevisiae
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4044 , http://hdl.handle.net/10962/d1004105 , Brewing -- Microbiology , Yeast , Fermentation , Saccharomyces cerevisiae
- Description: The brewing industry constantly experiences problems in trying to maintain the quality of beer produced. Unfavourable conditions during fermentation may alter the performance of the yeast strain Saccharomyces cerevisiae, resulting in a "poor" end-product. It has been established that high concentrations of extracellular ethanol, when added to the fermentation medium inhibit yeast activity. It has been recently suggested that increased carbon dioxide pressure could inactivate the yeast activity adding to further brewing problems. The aim of this study was to investigate the effect of extracellular carbon dioxide pressure and ethanol addition, on yeast performance when added to a fermentation medium, and to establish whether an inhibitory relationship existed between ethanol and carbon dioxide pressure, when combined and added to the fermentation medium. Dissolved C0₂ in the medium, medium pH and substrate utilisation were analysed daily during a fermentation, as were membrane fatty acid composition. These parameters were used to assess the effect of ethanol and carbon dioxide on the yeast performance and consequently the final end-product. Supplementing the medium with extracellular ethanol, even as low as 5%, was shown to inhibit yeast performance during fermentation. This effect was even more marked as the ethanol concentration was increased, with almost total inhibition of yeast activity occuring after the addition of 15% ethanol (v/v). A similar effect was observed when elevated C0₂ pressures were applied to the medium, and although low C0₂ pressures initially induced the synthesis of saturated yeast membrane fatty acids, elevated C0₂ pressures (greater than 1,0 atm.) was shown to follow a similar inhibitory trend, if not as dramatic, as ethanol. A combination of both ethanol and C0₂ pressure showed a further increase in the level of yeast inhibition, although the low C0₂ pressure appeared to initially inhibit the toxicity of ethanol on the yeast. Increasing the levels of the C0₂/ethanol treatment (1,0 atm.), showed a synergistic effect on yeast performance. The results of this study indicate that both extracellular ethanol and carbon dioxide do appear to inhibit yeast performance and affect membrane fatty acid composition of the cells by inhibiting the synthesis of the respective fatty acid. This affect has a significant bearing on the general metabolism of the yeast cell.
- Full Text:
- Date Issued: 1993
- Authors: Longden, Nicholas Guy
- Date: 1993
- Subjects: Brewing -- Microbiology , Yeast , Fermentation , Saccharomyces cerevisiae
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4044 , http://hdl.handle.net/10962/d1004105 , Brewing -- Microbiology , Yeast , Fermentation , Saccharomyces cerevisiae
- Description: The brewing industry constantly experiences problems in trying to maintain the quality of beer produced. Unfavourable conditions during fermentation may alter the performance of the yeast strain Saccharomyces cerevisiae, resulting in a "poor" end-product. It has been established that high concentrations of extracellular ethanol, when added to the fermentation medium inhibit yeast activity. It has been recently suggested that increased carbon dioxide pressure could inactivate the yeast activity adding to further brewing problems. The aim of this study was to investigate the effect of extracellular carbon dioxide pressure and ethanol addition, on yeast performance when added to a fermentation medium, and to establish whether an inhibitory relationship existed between ethanol and carbon dioxide pressure, when combined and added to the fermentation medium. Dissolved C0₂ in the medium, medium pH and substrate utilisation were analysed daily during a fermentation, as were membrane fatty acid composition. These parameters were used to assess the effect of ethanol and carbon dioxide on the yeast performance and consequently the final end-product. Supplementing the medium with extracellular ethanol, even as low as 5%, was shown to inhibit yeast performance during fermentation. This effect was even more marked as the ethanol concentration was increased, with almost total inhibition of yeast activity occuring after the addition of 15% ethanol (v/v). A similar effect was observed when elevated C0₂ pressures were applied to the medium, and although low C0₂ pressures initially induced the synthesis of saturated yeast membrane fatty acids, elevated C0₂ pressures (greater than 1,0 atm.) was shown to follow a similar inhibitory trend, if not as dramatic, as ethanol. A combination of both ethanol and C0₂ pressure showed a further increase in the level of yeast inhibition, although the low C0₂ pressure appeared to initially inhibit the toxicity of ethanol on the yeast. Increasing the levels of the C0₂/ethanol treatment (1,0 atm.), showed a synergistic effect on yeast performance. The results of this study indicate that both extracellular ethanol and carbon dioxide do appear to inhibit yeast performance and affect membrane fatty acid composition of the cells by inhibiting the synthesis of the respective fatty acid. This affect has a significant bearing on the general metabolism of the yeast cell.
- Full Text:
- Date Issued: 1993
Studies on the fermentation of molasses by Clostridium acetobutylicum
- Authors: Barber, Jennifer Mary
- Date: 1978
- Subjects: Molasses , Clostridium acetobutylicum , Fermentation
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4084 , http://hdl.handle.net/10962/d1007611 , Molasses , Clostridium acetobutylicum , Fermentation
- Description: The bacterium Clostridium acetobutylicum produces acetone and n [subscript] - butanol from molasses in an industrial fermentation system. Although the bacterium has been cultured in liquid media it does not grow well on agar plates and requires high concentrations of hydrogen. Pretreatment of agar plates with bovine catalase improves growth on agar media. The bacteria produce an area of clearing (halo) on Potato agar plates due to butyric acid (the precursor of n [subscript]-butanol) and ß -amylase production. This characteristic will be used as a plate screening assay for the selection of high solvent producing mutants. A laboratory scale fermentation system was developed and detailed studies including pH, turbidity and cell morphology changes, and the details of solvent production were undertaken. The fermentation was optimized for mutant selection. The production of normal solvent yields by isolated clones is required for the mutant selection programme. Studies revealed that sporulation of the clones increased their solvent yield although solvent yields were still lower than normal. Efficient sporulation is therefore a prerequisite for clone fermentation. The origin of the phage infection during the factory outbreak was determined and resistant clones obtained. The presence of a bacteriocin-like toxin causing decreases in turbidity was identified during the final fermentation stage. The strain sensitivity, optimum conditions for stability as well as the kinetics of inactivation and lethality have been investigated. Preliminary characterization and purification studies indicate the proteinaceous nature of the toxin. , KMBT_363
- Full Text:
- Date Issued: 1978
- Authors: Barber, Jennifer Mary
- Date: 1978
- Subjects: Molasses , Clostridium acetobutylicum , Fermentation
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4084 , http://hdl.handle.net/10962/d1007611 , Molasses , Clostridium acetobutylicum , Fermentation
- Description: The bacterium Clostridium acetobutylicum produces acetone and n [subscript] - butanol from molasses in an industrial fermentation system. Although the bacterium has been cultured in liquid media it does not grow well on agar plates and requires high concentrations of hydrogen. Pretreatment of agar plates with bovine catalase improves growth on agar media. The bacteria produce an area of clearing (halo) on Potato agar plates due to butyric acid (the precursor of n [subscript]-butanol) and ß -amylase production. This characteristic will be used as a plate screening assay for the selection of high solvent producing mutants. A laboratory scale fermentation system was developed and detailed studies including pH, turbidity and cell morphology changes, and the details of solvent production were undertaken. The fermentation was optimized for mutant selection. The production of normal solvent yields by isolated clones is required for the mutant selection programme. Studies revealed that sporulation of the clones increased their solvent yield although solvent yields were still lower than normal. Efficient sporulation is therefore a prerequisite for clone fermentation. The origin of the phage infection during the factory outbreak was determined and resistant clones obtained. The presence of a bacteriocin-like toxin causing decreases in turbidity was identified during the final fermentation stage. The strain sensitivity, optimum conditions for stability as well as the kinetics of inactivation and lethality have been investigated. Preliminary characterization and purification studies indicate the proteinaceous nature of the toxin. , KMBT_363
- Full Text:
- Date Issued: 1978
- «
- ‹
- 1
- ›
- »