Evaluation of pre-treatment methods on production of bioethanol from bagasse and sugarcane trash
- Authors: Dodo, Charlie Marembo
- Date: 2019
- Subjects: Lignocellulose
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10353/15387 , vital:40403
- Description: A variety of methods have been researched on for bioethanol preparation from different feedstocks. Amongst the available feedstock, one such feedstock is the sugarcane plant. In most of the research on bioethanol preparation with sugarcane the sugary juice has been widely used, with the bagasse and trash having been discarded as waste. The “waste” bagasse and trash are usually removed and thrown away or burnt during harvesting or in sugar mills to supplement energy requirements. This research on lignocellulosic bagasse and trash was done so as not to discard them but to rather find ways in which to use this biomass constructively. Alternatives to burning that can potentially add value to this biomass need to be researched on by evaluating their hydrolysis content. The different lignocellulose pretreatment methods of concentrated and dilute acid pretreatment, with subsequent enzyme hydrolysis as well as alkali and oxidative alkali pretreatment with enzyme hydrolysis were experimented on the bagasse and trash for hydrolysis efficiency and effectiveness. There are two types of acid hydrolysis which were investigated on which are concentrated and dilute sulphuric acid pretreatments. Use of concentrated sulphuric acid yielded the highest amounts of reducing sugars but also resulted in the highest amounts of downstream process inhibitors formation. This resulted in the need for neutralisation steps which in turn increase the overall costs of using this method to obtain reducing sugars. It has however the advantage of occurring at a faster rate, within minutes or hours, than using biological enzymes which took days, up to 72 hours to obtain the highest reducing sugar amounts. Dilute sulphuric acid pretreatment offered the advantage of using fewer chemicals which are therefore less severe on equipment and result in fewer fermentation inhibitors being formed. Dilute sulphuric acid hydrolysis also takes a relatively shorter period than biological methods of pretreatment. A challenge of fermentation inhibitors formed during acid hydrolysis was countered by using the methods of overliming (calcium hydroxide) and comparing it to neutralization with sodium hydroxide. Alkali pretreatment with sodium hydroxide was researched on by applying different pretreatment concentrations during experiments on the lignocellulosic biomass. There was an increase in the available quantities of cellulose with a significant reduction in lignin with pretreatment. Alkali pretreatment proved effective in exposing the cellulose which made v more cellulose surface area available to cellulase enzymes for enzyme hydrolysis. The highest yield of reducing sugars was obtained from hydrolysates pretreated with 0.25 M sodium hydroxide for 60 min and a period of 72 h of enzyme hydrolysis. In general the longer the pretreatment time the more reducing sugars were produced from the enzyme hydrolysis. Alkali peroxide pretreatment also resulted in significant reductions in lignin quantities of lignocellulose material. In this method sodium hydroxide in combination with hydrogen peroxide were used in pretreating the biomass. Hydrolysates with even fewer fermentation inhibitors were produced as a result. The highest percentage concentration of cellulose of 63% (g/g) was achieved after pretreatment of bagasse with 5% alkali hydrogen peroxide and trash with 0,25M sodium hydroxide pretreatment. Pretreatment of biomass using alkali with subsequent enzymatic hydrolysis gave the highest yields of fermentable sugars of 38% (g/g) using 7% (v/v) alkali peroxide pre-treated trash than 36% (g/g) for 5% (v/v) with the least inhibitors. Reducing sugar yields of 25% (g/g) and 22% (g/g) were obtained after pretreatment with concentrated and dilute acid respectively. Neutralization of the acid hydrolysates was necessary to reduce inhibitors formed with neutralisation by sodium hydroxide resulting in low dilutions and loss of fermentable sugars as unlike in the case of overliming. Subsequent steps of fermenting the reducing sugars resulting from pretreatment into bioethanol were based on using the yeast Saccharomyces cerevisae. Pretreatment hydrolysates from alkali peroxide experiments produced higher bioethanol yields of 13.7 (g/l) after enzyme hydrolysates versus 6.9 (g/l) bioethanol from dilute acid hydrolyzates. A comparison of the effects of time showed there was more bioethanol yield of 13.7 (g/l) after 72 h of fermentation with the yeast versus 7.0 (g/l) bioethanol after pretreatment for 24 h. The only drawback is the longer fermentation period which thus reduces the process and so reduces the value of the increase in yield
- Full Text:
- Date Issued: 2019
- Authors: Dodo, Charlie Marembo
- Date: 2019
- Subjects: Lignocellulose
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10353/15387 , vital:40403
- Description: A variety of methods have been researched on for bioethanol preparation from different feedstocks. Amongst the available feedstock, one such feedstock is the sugarcane plant. In most of the research on bioethanol preparation with sugarcane the sugary juice has been widely used, with the bagasse and trash having been discarded as waste. The “waste” bagasse and trash are usually removed and thrown away or burnt during harvesting or in sugar mills to supplement energy requirements. This research on lignocellulosic bagasse and trash was done so as not to discard them but to rather find ways in which to use this biomass constructively. Alternatives to burning that can potentially add value to this biomass need to be researched on by evaluating their hydrolysis content. The different lignocellulose pretreatment methods of concentrated and dilute acid pretreatment, with subsequent enzyme hydrolysis as well as alkali and oxidative alkali pretreatment with enzyme hydrolysis were experimented on the bagasse and trash for hydrolysis efficiency and effectiveness. There are two types of acid hydrolysis which were investigated on which are concentrated and dilute sulphuric acid pretreatments. Use of concentrated sulphuric acid yielded the highest amounts of reducing sugars but also resulted in the highest amounts of downstream process inhibitors formation. This resulted in the need for neutralisation steps which in turn increase the overall costs of using this method to obtain reducing sugars. It has however the advantage of occurring at a faster rate, within minutes or hours, than using biological enzymes which took days, up to 72 hours to obtain the highest reducing sugar amounts. Dilute sulphuric acid pretreatment offered the advantage of using fewer chemicals which are therefore less severe on equipment and result in fewer fermentation inhibitors being formed. Dilute sulphuric acid hydrolysis also takes a relatively shorter period than biological methods of pretreatment. A challenge of fermentation inhibitors formed during acid hydrolysis was countered by using the methods of overliming (calcium hydroxide) and comparing it to neutralization with sodium hydroxide. Alkali pretreatment with sodium hydroxide was researched on by applying different pretreatment concentrations during experiments on the lignocellulosic biomass. There was an increase in the available quantities of cellulose with a significant reduction in lignin with pretreatment. Alkali pretreatment proved effective in exposing the cellulose which made v more cellulose surface area available to cellulase enzymes for enzyme hydrolysis. The highest yield of reducing sugars was obtained from hydrolysates pretreated with 0.25 M sodium hydroxide for 60 min and a period of 72 h of enzyme hydrolysis. In general the longer the pretreatment time the more reducing sugars were produced from the enzyme hydrolysis. Alkali peroxide pretreatment also resulted in significant reductions in lignin quantities of lignocellulose material. In this method sodium hydroxide in combination with hydrogen peroxide were used in pretreating the biomass. Hydrolysates with even fewer fermentation inhibitors were produced as a result. The highest percentage concentration of cellulose of 63% (g/g) was achieved after pretreatment of bagasse with 5% alkali hydrogen peroxide and trash with 0,25M sodium hydroxide pretreatment. Pretreatment of biomass using alkali with subsequent enzymatic hydrolysis gave the highest yields of fermentable sugars of 38% (g/g) using 7% (v/v) alkali peroxide pre-treated trash than 36% (g/g) for 5% (v/v) with the least inhibitors. Reducing sugar yields of 25% (g/g) and 22% (g/g) were obtained after pretreatment with concentrated and dilute acid respectively. Neutralization of the acid hydrolysates was necessary to reduce inhibitors formed with neutralisation by sodium hydroxide resulting in low dilutions and loss of fermentable sugars as unlike in the case of overliming. Subsequent steps of fermenting the reducing sugars resulting from pretreatment into bioethanol were based on using the yeast Saccharomyces cerevisae. Pretreatment hydrolysates from alkali peroxide experiments produced higher bioethanol yields of 13.7 (g/l) after enzyme hydrolysates versus 6.9 (g/l) bioethanol from dilute acid hydrolyzates. A comparison of the effects of time showed there was more bioethanol yield of 13.7 (g/l) after 72 h of fermentation with the yeast versus 7.0 (g/l) bioethanol after pretreatment for 24 h. The only drawback is the longer fermentation period which thus reduces the process and so reduces the value of the increase in yield
- Full Text:
- Date Issued: 2019
The microbial ecology of sulphidogenic lignocellulose degradation
- Authors: Clarke, Anna Maria
- Date: 2007
- Subjects: Microbial ecology , Lignocellulose , Sulfides , Lignin , Lignocellulose -- Biodegradation , Mines and mineral resources -- Waste disposal , Acid mine drainage
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4094 , http://hdl.handle.net/10962/d1008181
- Description: Acid mine drainage is a well known environmental pollutant, not only in South Africa, but throughout the world, and the use of microbial processes in the treatment of these wastes has been the subject of investigation over past decades. Lignocellulose packed-bed reactors have been used in passive treatment systems, and, although effective initially, they show early decline in performance while the packing material remains largely un-utilized. Little is known about this phenomenon which remains a severe constraint in the development of efficient passive mine water treatment systems. It has been proposed that the degradation pathways of the complex lignocellulose substrate may be limited in some way in these systems during the manifestation of this effect. This study has addressed the problem using a molecular microbial ecology methodology in an attempt to relate trophic functions of the microbial population to the physico-chemical data of the system. A field-scale lignocellulose packed-bed reactor located at Vryheid Coronation Colliery (Northern Kwa-Zulu Natal province, South Africa) was monitored for six years and the results showed the classic profile of performance decline related to a slowdown in sulphate reduction and alkalinity production. The reactor was decommissioned , comprehensive samples were collected along the depth profile and the microbial populations investigated by means of 16S rRNA gene methodology. The population was found to include cellulolytic Clostridia spp., CytophagaIFlavobacterlBacteroidetes, Sphingomonadaceae and as yet uncultured microorganisms related to microbiota identified in the rumen and termite gut. These are all known to be involved as primary fermenters of cellulose. Oesulphosporosinus was present as sulphate reducer. A comparison of substrata sampling and population distribution suggested that spatial and temporal gradients within the system may become established over the course of its operation. Based on these findings, a laboratory-scale reactor was constructed to simulate the performance of the packed-bed reactor under controlled experimental conditions. The laboratory-scale reactor was operated for 273 days and showed comparable performance to that in the field in both biomolecular and physicochemical data. Clearly defined trophic niches were observed. These results suggested that a sequence of events does occur in lignocellulose degradation over time. Based on the spatial and temporal column studies, a descriptive model was proposed to account for these events. It was found that fermentative organisms predominate in the inlet zone of the system using easily extractable compounds from the wood, thus providing feedstock for sulphate reduction occurring in the succeeding compartments. Production of sulphide and alkalinity appears to be involved in the enhancement of lignin degradation and this, in turn, appears to enhance access to the cellulose fraction. However, once the readily extractables are exhausted, the decline in sulphide and alkalinity production leads inexorably to a decline in the overall performance of the system as a sulphate reducing unit operation. These observations led to the proposal that with the addition of a limited amount of a readily available carbon source, such as molasses, in the initial zone of the the reactor, the ongoing generation of sulphide would be sustained and this in turn would sustain the microbial attack on the lignocellulose complex. This proposal was tested in scale-up studies and positive results indicate that the descriptive model may, to some extent, provide an account of events occurring in these systems. The work on sustaining lignocellulose degradation through the maintenance of sulphate reduction in the initial stages of the reactor flow path has led to the development of the Degrading Packed-bed Reactor concept and that, has subsequently been successfully evaluated in the field.
- Full Text:
- Date Issued: 2007
- Authors: Clarke, Anna Maria
- Date: 2007
- Subjects: Microbial ecology , Lignocellulose , Sulfides , Lignin , Lignocellulose -- Biodegradation , Mines and mineral resources -- Waste disposal , Acid mine drainage
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: vital:4094 , http://hdl.handle.net/10962/d1008181
- Description: Acid mine drainage is a well known environmental pollutant, not only in South Africa, but throughout the world, and the use of microbial processes in the treatment of these wastes has been the subject of investigation over past decades. Lignocellulose packed-bed reactors have been used in passive treatment systems, and, although effective initially, they show early decline in performance while the packing material remains largely un-utilized. Little is known about this phenomenon which remains a severe constraint in the development of efficient passive mine water treatment systems. It has been proposed that the degradation pathways of the complex lignocellulose substrate may be limited in some way in these systems during the manifestation of this effect. This study has addressed the problem using a molecular microbial ecology methodology in an attempt to relate trophic functions of the microbial population to the physico-chemical data of the system. A field-scale lignocellulose packed-bed reactor located at Vryheid Coronation Colliery (Northern Kwa-Zulu Natal province, South Africa) was monitored for six years and the results showed the classic profile of performance decline related to a slowdown in sulphate reduction and alkalinity production. The reactor was decommissioned , comprehensive samples were collected along the depth profile and the microbial populations investigated by means of 16S rRNA gene methodology. The population was found to include cellulolytic Clostridia spp., CytophagaIFlavobacterlBacteroidetes, Sphingomonadaceae and as yet uncultured microorganisms related to microbiota identified in the rumen and termite gut. These are all known to be involved as primary fermenters of cellulose. Oesulphosporosinus was present as sulphate reducer. A comparison of substrata sampling and population distribution suggested that spatial and temporal gradients within the system may become established over the course of its operation. Based on these findings, a laboratory-scale reactor was constructed to simulate the performance of the packed-bed reactor under controlled experimental conditions. The laboratory-scale reactor was operated for 273 days and showed comparable performance to that in the field in both biomolecular and physicochemical data. Clearly defined trophic niches were observed. These results suggested that a sequence of events does occur in lignocellulose degradation over time. Based on the spatial and temporal column studies, a descriptive model was proposed to account for these events. It was found that fermentative organisms predominate in the inlet zone of the system using easily extractable compounds from the wood, thus providing feedstock for sulphate reduction occurring in the succeeding compartments. Production of sulphide and alkalinity appears to be involved in the enhancement of lignin degradation and this, in turn, appears to enhance access to the cellulose fraction. However, once the readily extractables are exhausted, the decline in sulphide and alkalinity production leads inexorably to a decline in the overall performance of the system as a sulphate reducing unit operation. These observations led to the proposal that with the addition of a limited amount of a readily available carbon source, such as molasses, in the initial zone of the the reactor, the ongoing generation of sulphide would be sustained and this in turn would sustain the microbial attack on the lignocellulose complex. This proposal was tested in scale-up studies and positive results indicate that the descriptive model may, to some extent, provide an account of events occurring in these systems. The work on sustaining lignocellulose degradation through the maintenance of sulphate reduction in the initial stages of the reactor flow path has led to the development of the Degrading Packed-bed Reactor concept and that, has subsequently been successfully evaluated in the field.
- Full Text:
- Date Issued: 2007
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