Development of bio-based xylan composites for food packaging applications
- Authors: Naidu, Darrel Sarvesh
- Date: 2020
- Subjects: Xylanases
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/48544 , vital:40886
- Description: Currently a large number of chemicals and plastics are produced from petroleum-based resources. However, due to the concerns surrounding the depletion of petroleum resources and growing carbon emissions, there is a desire to produce chemicals and plastics from renewable and carbon natural sources. Lignocellulosic biomass (biomass consisting of cellulose, lignin and hemicellulose) is the most common biomass on earth and is renewable and carbon neutral. Of the three main constituents of lignocellulosic biomass, hemicellulose is composed of a mixture of sugars which can be converted into chemicals and plastics. The most common form of hemicellulose found in nature is xylan. This study is aimed at extracting xylan from maize stalk waste residues and the development of xylan films with properties that are suitable for food packaging applications. Xylan was extracted from maize stalk waste residues using an alkaline pre-treatment method. The effects of bleaching conditions (time, temperature and bleach concentration) prior to alkaline treatment on the yield and purity of xylan extracted was studied using a Box-Behnken experimental design. It was observed that the experimental conditions tested had no effect on the yield of the xylan extracted but the bleach concentration had a significant effect on the purity of the xylan extracted. The samples with the lowest lignin content were found to be the samples treated with 3wt% bleach prior to alkaline pre-treatment. One of the main disadvantages of xylan is that it has poor film forming properties, a method of overcoming this is to combine it with another abundantly available biopolymer that has good film forming properties, such as alginate. The effect of xylan, alginate and glycerol content on the mechanical, thermal, moisture uptake and water barrier properties of the films were investigated. It was observed that with an increase in the alginate content there was an increase in the tensile strength and Young’s modulus of the films, whereas the water vapour permeability (WVP) of the films decreased. This was attributed to the greater cohesion between alginate polymer chains compared to the cohesion between xylan polymer chains. The xylanalginate films exhibited better optical and water sorption properties at higher xylan content. In order to improve the inherently poor mechanical and barrier properties of the xylan-alginate films, bentonite and halloysite were incorporated into the films. It was found that 5wt% incorporation of either bentonite or halloysite resulted in a 49% decrease of the WVP, which was attributed to water vapour impermeable nature of the silicate layers that make up both clays. The incorporation of the clays into the xylan-alginate matrix resulted in a significant
- Full Text:
- Date Issued: 2020
- Authors: Naidu, Darrel Sarvesh
- Date: 2020
- Subjects: Xylanases
- Language: English
- Type: Thesis , Doctoral , PhD
- Identifier: http://hdl.handle.net/10948/48544 , vital:40886
- Description: Currently a large number of chemicals and plastics are produced from petroleum-based resources. However, due to the concerns surrounding the depletion of petroleum resources and growing carbon emissions, there is a desire to produce chemicals and plastics from renewable and carbon natural sources. Lignocellulosic biomass (biomass consisting of cellulose, lignin and hemicellulose) is the most common biomass on earth and is renewable and carbon neutral. Of the three main constituents of lignocellulosic biomass, hemicellulose is composed of a mixture of sugars which can be converted into chemicals and plastics. The most common form of hemicellulose found in nature is xylan. This study is aimed at extracting xylan from maize stalk waste residues and the development of xylan films with properties that are suitable for food packaging applications. Xylan was extracted from maize stalk waste residues using an alkaline pre-treatment method. The effects of bleaching conditions (time, temperature and bleach concentration) prior to alkaline treatment on the yield and purity of xylan extracted was studied using a Box-Behnken experimental design. It was observed that the experimental conditions tested had no effect on the yield of the xylan extracted but the bleach concentration had a significant effect on the purity of the xylan extracted. The samples with the lowest lignin content were found to be the samples treated with 3wt% bleach prior to alkaline pre-treatment. One of the main disadvantages of xylan is that it has poor film forming properties, a method of overcoming this is to combine it with another abundantly available biopolymer that has good film forming properties, such as alginate. The effect of xylan, alginate and glycerol content on the mechanical, thermal, moisture uptake and water barrier properties of the films were investigated. It was observed that with an increase in the alginate content there was an increase in the tensile strength and Young’s modulus of the films, whereas the water vapour permeability (WVP) of the films decreased. This was attributed to the greater cohesion between alginate polymer chains compared to the cohesion between xylan polymer chains. The xylanalginate films exhibited better optical and water sorption properties at higher xylan content. In order to improve the inherently poor mechanical and barrier properties of the xylan-alginate films, bentonite and halloysite were incorporated into the films. It was found that 5wt% incorporation of either bentonite or halloysite resulted in a 49% decrease of the WVP, which was attributed to water vapour impermeable nature of the silicate layers that make up both clays. The incorporation of the clays into the xylan-alginate matrix resulted in a significant
- Full Text:
- Date Issued: 2020
Lignocellulosic waste degradation using enzyme synergy with commercially available enzymes and Clostridium cellulovorans XylanaseA and MannanaseA
- Authors: Morrison, David Graham
- Date: 2014
- Subjects: Lignocellulose -- Biodegradation , Enzymes -- Biotechnology , Agricultural wastes as fuel , Polysaccharides -- Biotechnology , Sugar -- Inversion , Clostridium , Xylanases , Monomers
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4119 , http://hdl.handle.net/10962/d1013292
- Description: The launch of national and international initiatives to reduce pollution, reliance on fossil fuels and increase the beneficiation of agricultural wastes has prompted research into sugar monomer production from lignocellulosic wastes. These sugars can subsequently be used in the production of biofuels and environmentally degradable plastics. This study investigated the use of synergistic combinations of commercial and pure enzymes to lower enzyme costs and loadings, while increasing enzyme activity in the hydrolysis of agricultural waste. Pineapple pomace was selected due to its current underutilisation and the substantial quantities of it produced annually, as a by-product of pineapple canning. One of the primary costs in beneficiating agricultural wastes, such as pineapple pomace, is the high cost of enzyme solutions used to generate reducing sugars. This can be lowered through the use of synergistic combinations of enzymes. Studies related to the inclusion of hemicellulose degrading enzymes with commercial enzyme solutions have been limited and investigation of these solutions in select combinations, together with pineapple pomace substrate, allows for novel research. The use of synergistic combinations of purified cellulosomal enzymes has previously been shown to be effective at releasing reducing sugars from agricultural wastes. For the present study, MannanaseA and XylanaseA from Clostridium cellulovorans were heterologously expressed in Escherichia coli BL21 (DE3) cells and purified with immobilised metal affinity chromatography. These enzymes, in addition to two commercially available enzyme solutions (Celluclast 1.5L® and Pectinex® 3XL), were assayed on defined polysaccharides that are present in pineapple pomace to determine their substrate specificities. The degree(s) of synergy and specific activities of selected combinations of these enzymes were tested under both simultaneous and sequential conditions. It was observed that several synergistic combinations of enzyme solutions in select ratios, such as C20P60X20 (20% cellulose, 60% pectinase and 20% xylanse), C20P40X40 (20% cellulose, 40% pectinase and 40% xylanase) and C20P80 (20% cellulose, 80% pectinase) with pineapple pomace could both decrease the protein loading, while raising the level of activity compared to individual enzyme solutions. The highest quantity of reducing sugars to protein weight used on pineapple pomace was recorded at 3, 9 and 18 hours with combinations of Pectinex® 3XL and Celluclast 1.5L®, but for 27 h it was combinations of both these commercial solutions with XynA. The contribution of XynA was significant as C20P60X20 displayed the second highest reducing sugar production of 1.521 mg/mL, at 36 h from 12.875 μg/mL of protein, which was the second lowest protein loading. It was also shown that certain enzyme combinations, such as Pectinex® 3XL, Celluclast 1.5L® and XynA, did not generate synergy when combined in solution at the initial stages of hydrolysis, and instead generated a form of competition called anti-synergy. This was due to Pectinex® 3XL which had anti-synergy relationships in select combinations with the other enzyme solutions assayed. It was also observed that the degree of synergy and specific activity for a combination changed over time. Some solutions displayed the highest levels of synergy at the commencement of hydrolysis, namely Celluclast 1.5L®, ManA and XynA. Other combinations exhibited the highest levels of synergy at the end of the assay period, such as Pectinex® 3XL and Celluclast 1.5L®. Whether greater synergy was generated at the start or end of hydrolysis was a function of the stability of the enzymes in solution and whether enzyme activity increased substrate accessibility or generated competition between enzymes in solution. Sequential synergy studies demonstrated an anti-synergy relationship between Pectinex® 3XL and XynA or ManA, as well as Pectinex® 3 XL and Celluclast 1.5L®. It was found that under sequential synergy conditions with Pectinex® 3 XL, XynA and ManA, that anti-synergy could be negated and high degrees of synergy attained when the enzymes were added in specific loading orders and not inhibited by the presence of other active enzymes. The importance of loading order was demonstrated under sequential synergy conditions when XynA was added before ManA followed by Pectinex® 3 XL, which increased the activity and synergy of the solution by 50%. This equates to a 60% increase in reducing sugar release from the same concentrations of enzymes and emphasises the importance of removing anti-synergy relationships from combinations of enzymes. It can be concluded that a C20P60X20 combination (based on activity) can both synergistically increase the reducing sugar production and lower the protein loading required for pineapple pomace hydrolysis. This study also highlights the importance of reducing anti-synergy in customised enzyme cocktails and how sequential synergy can demonstrate the order in which a lignocellulosic waste is degraded.
- Full Text:
- Date Issued: 2014
- Authors: Morrison, David Graham
- Date: 2014
- Subjects: Lignocellulose -- Biodegradation , Enzymes -- Biotechnology , Agricultural wastes as fuel , Polysaccharides -- Biotechnology , Sugar -- Inversion , Clostridium , Xylanases , Monomers
- Language: English
- Type: Thesis , Masters , MSc
- Identifier: vital:4119 , http://hdl.handle.net/10962/d1013292
- Description: The launch of national and international initiatives to reduce pollution, reliance on fossil fuels and increase the beneficiation of agricultural wastes has prompted research into sugar monomer production from lignocellulosic wastes. These sugars can subsequently be used in the production of biofuels and environmentally degradable plastics. This study investigated the use of synergistic combinations of commercial and pure enzymes to lower enzyme costs and loadings, while increasing enzyme activity in the hydrolysis of agricultural waste. Pineapple pomace was selected due to its current underutilisation and the substantial quantities of it produced annually, as a by-product of pineapple canning. One of the primary costs in beneficiating agricultural wastes, such as pineapple pomace, is the high cost of enzyme solutions used to generate reducing sugars. This can be lowered through the use of synergistic combinations of enzymes. Studies related to the inclusion of hemicellulose degrading enzymes with commercial enzyme solutions have been limited and investigation of these solutions in select combinations, together with pineapple pomace substrate, allows for novel research. The use of synergistic combinations of purified cellulosomal enzymes has previously been shown to be effective at releasing reducing sugars from agricultural wastes. For the present study, MannanaseA and XylanaseA from Clostridium cellulovorans were heterologously expressed in Escherichia coli BL21 (DE3) cells and purified with immobilised metal affinity chromatography. These enzymes, in addition to two commercially available enzyme solutions (Celluclast 1.5L® and Pectinex® 3XL), were assayed on defined polysaccharides that are present in pineapple pomace to determine their substrate specificities. The degree(s) of synergy and specific activities of selected combinations of these enzymes were tested under both simultaneous and sequential conditions. It was observed that several synergistic combinations of enzyme solutions in select ratios, such as C20P60X20 (20% cellulose, 60% pectinase and 20% xylanse), C20P40X40 (20% cellulose, 40% pectinase and 40% xylanase) and C20P80 (20% cellulose, 80% pectinase) with pineapple pomace could both decrease the protein loading, while raising the level of activity compared to individual enzyme solutions. The highest quantity of reducing sugars to protein weight used on pineapple pomace was recorded at 3, 9 and 18 hours with combinations of Pectinex® 3XL and Celluclast 1.5L®, but for 27 h it was combinations of both these commercial solutions with XynA. The contribution of XynA was significant as C20P60X20 displayed the second highest reducing sugar production of 1.521 mg/mL, at 36 h from 12.875 μg/mL of protein, which was the second lowest protein loading. It was also shown that certain enzyme combinations, such as Pectinex® 3XL, Celluclast 1.5L® and XynA, did not generate synergy when combined in solution at the initial stages of hydrolysis, and instead generated a form of competition called anti-synergy. This was due to Pectinex® 3XL which had anti-synergy relationships in select combinations with the other enzyme solutions assayed. It was also observed that the degree of synergy and specific activity for a combination changed over time. Some solutions displayed the highest levels of synergy at the commencement of hydrolysis, namely Celluclast 1.5L®, ManA and XynA. Other combinations exhibited the highest levels of synergy at the end of the assay period, such as Pectinex® 3XL and Celluclast 1.5L®. Whether greater synergy was generated at the start or end of hydrolysis was a function of the stability of the enzymes in solution and whether enzyme activity increased substrate accessibility or generated competition between enzymes in solution. Sequential synergy studies demonstrated an anti-synergy relationship between Pectinex® 3XL and XynA or ManA, as well as Pectinex® 3 XL and Celluclast 1.5L®. It was found that under sequential synergy conditions with Pectinex® 3 XL, XynA and ManA, that anti-synergy could be negated and high degrees of synergy attained when the enzymes were added in specific loading orders and not inhibited by the presence of other active enzymes. The importance of loading order was demonstrated under sequential synergy conditions when XynA was added before ManA followed by Pectinex® 3 XL, which increased the activity and synergy of the solution by 50%. This equates to a 60% increase in reducing sugar release from the same concentrations of enzymes and emphasises the importance of removing anti-synergy relationships from combinations of enzymes. It can be concluded that a C20P60X20 combination (based on activity) can both synergistically increase the reducing sugar production and lower the protein loading required for pineapple pomace hydrolysis. This study also highlights the importance of reducing anti-synergy in customised enzyme cocktails and how sequential synergy can demonstrate the order in which a lignocellulosic waste is degraded.
- Full Text:
- Date Issued: 2014
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