Thesis (Ph. D.)--Institute of Paper Chemistry, 1941. / Bibliography: leaves 60-61.

Pre-treatment of wheat straw(lignocellulosic) biomass is a crucial step as it has direct impact on the subsequent yield of enzymatic saccharification and alcohol fermentation processes in the production of biofuel. Twin screw extrusion is a highly feasible pretreatment method and has been received great interest in the recent year pre-treatment studies. Twin screw extrusion is a continuous process, where the biomass feedstock can be subjected to a combination of simultaneous physical, thermal and chemical treatments. Steam explosion is a batch process and is the most commonly used method for lignocellulosic pre-treatment. In the initial stage of this study, the yield of glucose obtained from enzymatic saccharification for both methods (extrusion and steam explosion) were compared to identify the most effective pre-treatment approach. Effectiveness of the conventional steam explosion pre-treatment was used as benchmark for the directions of development of effective extrusion fractionation for wheat straw. In subsequent study, the impact of physical operating parameters (moisture, barrel temperature, compaction, screw speed and size reduction before extrusion) over twin screw extrusion with and without NaOH were studied. Low temperature (50°C) and increased moisture extrusion were preferred extrusion conditions. Yield of glucose can be improved by addition of NaOH (0.04g / g straw) and barrel temperature profile optimisation. Post extrusion washing was recommended. Findings from FTIR and TGA help to understand the chemical and structural changes took place in the pre-treatment and can be correlated with the glucose yield at the end of enzymatic hydrolysis. Characterisation analysis was extended to FT-NIR, morphology, crystallinity and specific surface area analysis to analyse the structural changes of lignocellulose biomass in extrusion pre-treatment and correlation with glucose yield. Chemometric analysis was used to statistically process large amounts of spectral data. The PCA scores plots showed good cluster segregation of the samples and were thus able to distinguish the effects of different pre-treatment conditions. The PLS regression models for both FTIR and FT-NIR showed good statistical regression and predictive ability correlated to the glucose yield. For the lignin ultilisation study, crude lignin was recovered from black liquor and fractionated with solvents. Lignin and the fractions were characterised with solvent solubility, SEC, UV, FTIR, 1H and 13C NMR and evaluated for the antioxidant activity with AAI ranged from 0.3 to 2.4. Reason for the low performance was proposed and experiment was extended to the intended application performance screening. Lignin application study was further extended to assess the feasibility of using lignin as an antioxidant in carboxylated acrilonitrile-butadiene rubber, XNBR glove. Evaluation involved physical observation, mechanical properties and thermal analysis – DSC-OIT after incorporation of lignin into XNBR glove. Lignin antioxidant performance was compared with current chemical antioxidant in used in industry. A part from antioxidant behaviour, lignin was also found can enhance the softness of XNBR film after accelerated heat aging.

Lignin is a major chemical component of plants and the second most abundant natural polymer after cellulose. The concerns and interests of agriculture and industry have stimulated the study of genes governing lignin content in plants in an effort to adapt plants to human purposes. Arabidopsis thaliana provides a convenient model for the study of the genes governing lignin content because of its short growth cycle, small plant size, and small completely sequenced genome. In order to identify the genes controlling lignin content in Arabidopsis accessions using Quantitative Trait Locus (QTL) analysis, a rapid micro-scale method of lignin determination is required. The acetyl bromide method has been modified to enable the rapid micro-scale determination of lignin content in Arabidopsis. Modifications included the use of a micro-ball mill, adoption of a modified rapid method of extraction, use of an ice-bath to stabilize solutions and reduction in solution volumes. The modified method was shown to be accurate and precise with values in agreement with those determined by the conventional method. The extinction coefficient for Arabidopsis lignin, dissolved using acetyl bromide, was determined to be 23.35 g-iLcm-1. This value is independent of the Arabidopsis accession, environmental growth conditions and is insensitive to syringyl/guaiacyl ratio. The modified acetyl bromide method was shown to be well correlated with the 72% sulfuric acid method once the latter had been corrected for protein contamination and acid-soluble lignin content (R² = 0.988, P < 0.0001). As determined by the newly developed acetyl bromide method and confirmed by the sulfuric acid method, lignin content in Arabidopsis was found to be a divergent property. Lignin content in Arabidopsis was found to be weekly correlated with growth rate among Arabidopsis accessions (R² = 0.48, P = 0.011). Lignin content was also found to be correlated with plant height among Arabidopsis accessions (R² = 0.491, P < 0.0001).

Currently, creosote-treated wood waste is mainly landfilled but the Province of Nova Scotia would prefer to see the waste converted to a valuable product. Bioremediation provides a treatment option for creosote-treated wood waste and the production of a value-added product that would have economic and environmental benefits. Composting technique can be used to degrade the contaminants and convert wood waste into humus and nutrients rich product and reduces the waste volume. The aim of this study was, therefore, to test the efficiency of the composting process in degrading the creosote in the contaminated wood waste. A laboratory scale bioremediation process was carried out in three specially designed in-vessel bioreactors equipped with thermo-insulating outer layer, mixing units, controlled aeration units, thermocouples a data logger and a computer. The three bioreactors were used as replicates. The ability of three thermophilic and cellulolytic microorganisms (T. curvata, T. aurantiacus and T. fusca) to degrade lignocellulose substrate was tested.

Textile dyes are molecules designed to impart a permanent colour to textile fabrics. They pose an environmental problem because they are toxic and they decrease the aesthetic value of rivers and lakes. Current technologies for dye removal cannot remove all classes of dyes and two or more technologies are usually combined to achieve statisfactory decolourization efficiencies. Lignin-degrading enzymes like laccases are potential technologies for dye decolourization and decolourization with immobilized laccase has been intensively investigated. The majority of those studies however have focused on dye disappearance and several reported that significant dye adsorption had occured during the dye removal, making the role of the enzyme unclear. Moreover, textile wastewaters contain auxiliary chemicals that can impact enzymatic dye decolourization and very few studies have evaluated the impact of those substances on laccase. This research evaluated the feasibility of treating dye-contaminated textile wastewaters with an immobilized laccase system. The first sub-objective was to examined the decolourization of Reactive blue 19 (an anthraquinone dye) by Trametes versicolor laccase immobilized on controlled porosity carrier (CPC) silica beads and the second was to analyze the kinetic effects of a non-ionic surfactant Merpol, sodium sulfate, and sodium chloride on laccase decolourization of Reactive blue 19. Decolourization of Reactive blue 19 by immobilized laccase was mainly enzymatic although dye some adsorption occurred. Decolourization led to less toxic by-products from azo and indigoid dyes whereas increased toxicity was observed for anthraquinone dyes. The feasibility of immobilizing laccase on poly(methyl methacrylate) (PMMA) through its sugar residues with a simple procedure was demonstrated and the mass of enzyme immobilized compared well with other commercial acrylic supports. The decolorization of Reactive blue 19 by laccase was inhibited by the non-ionic surfactant, Merpol by substrate depletion. A model describing this inhibition was developed and was validated by a saturated equilibrium binding experiment. While sodium sulfate (ionic strength) had no effect on either ABTS oxidation or dye decolourization, sodium chloride inhibited laccase during dye decolourization and the type and nature of the inhibition depended on the substrate. With ABTS, the inhibition was hyperbolic non-competitive whereas it was parabolic mixed with Reactive blue 19. / Thesis (Ph.D, Chemical Engineering) -- Queen's University, 2009-06-16 16:58:47.753

Two trials were conducted in order to assess the efficacy of microbial phytase supplementation in improving the bioavailability of certain nutrients in the diet of early weaned piglets. In trial I, a low-phosphorus, corn-soybean meal diet supplemented with phytase significantly improved the apparent digestibilities of phosphorus (P, p = 0.0004) and calcium (Ca, p = 0.04). The average daily gain (ADG) and feed conversion ratio (FCR) were also improved by supplemental phytase. In trial II, addition of acetic acid to the low-P plus phytase diet further improved the apparent digestibilities of P and Ca (gp = 0.01, p = 0.02, respectively). Furthermore, the apparent digestibility of copper was also significantly improved by acetic acid addition. As a consequence, the piglets fed this treatment showed the best growth performance. Acidification of a diet which provided the NRC recommended levels of available P significantly improved the apparent digestibilities of protein and most of the minerals (P, Fe, Zn, Cu and Mn) but it was not reflected in improved growth performance. These results indicate that microbial phytase not only released P from the phytic acid molecule but it also cleaved other minerals bound to this complex.

The potential of lignin and mannanoligosaccharides (MOS), as alternatives to antibiotic growth promoters was evaluated in broilers. Dietary treatments included: (1) negative control (CTL-, antibiotic free); (2) positive control (CTL+, 11 mg/kg virginiamycin); (3) MOS (diet 1 + Bio-Mos: 0.2% to 21 d and 0.1% thereafter); (4) LL (diet 1 + 1.25% Alcell lignin); (5) HL (diet 1 + 2.5% Alcell lignin). Bodyweight and feed conversion were not different when broilers were fed the CTL+, MOS, LL or HL diet. Birds fed MOS or LL had increased jejunum villi height (P &lt; 0.05) and greater goblet cell number per villus (P &lt; 0.05) when compared to those fed the CTL+ diet. MOS and LL increased (P &lt; 0.05) the cecal populations of Lactobacilli and Bifidobacteria when compared to CTL+ fed birds. However, Lactobacilli and Bifidobacteria loads were lowest (P &lt; 0.05) in birds fed the CTL+ or HL diet. Litter E. coli load was reduced (P &lt; 0.05) when birds were fed MOS than when fed the CTL+ diet, but comparable to LL or HL fed birds. In birds challenged with pathogenic strains of E. coli (O2 and O88 serotypes) and fed the MOS or HL diet, the cecal population of total E. coli was lower (P &lt; 0.05) than those fed the CTL+ diet; LL fed birds tended to have lower E. coli load than CTL+ fed birds. In summary, birds fed the MOS or LL diet had comparative advantage over CTL+ fed birds as evidenced by increased cecal populations of Lactobacilli and Bifdobacteria, increased villi height and greater goblet cell number in the jejunum, lower E. coli load in the litter, and lower cecal population of E. coli after an in vivo challenge with pathogenic strains of E. coli. Therefore, MOS and lignin could be regarded as natural alternatives to antibiotic growth promoters in poultry production. / Key words. Antibiotics, mannanoligosaccharides, lignin, gut health, broilers.

Under cellulolytic conditions, the white-rot fungus Trametes versicolor produces cellobiose dehydrogenase (CDH), an enzyme with a number of biochemical properties that are potentially relevant to the degradation of lignin and cellulose. To clarify its biochemical properties, CDH was purified from cultures of T. versicolor. Two isoforms of CDH were found: a 97 kDa isoform with both heme and flavin cofactors, and an 81 kDa isoform with a flavin cofactor. Both isoforms of CDH were found to be quite non-specific in their reductive half reactions. The flavin enzyme catalyzed many of the same reactions as the heme/flavin enzyme, but less efficiently. The flavin isoform reduced Fe(III) and Cu(II) only at concentrations well above those found physiologically. Thus the heme/flavin enzyme, but not the flavin enzyme, could be involved in promoting and sustaining the generation of hydroxyl radicals (&middot;OH) by Fenton's chemistry. / To characterize further the structural features of CDH, a genomic clone was isolated and sequenced. CDH was found to consist of 748 amino acids, without its predicted 19 amino acid signal peptide. Consistent with the domain structure of other CDHs, T. versicolor CDH appeared to be divided into an amino terminal heme domain and a carboxy terminal flavin domain, connected by a hydroxyamino acid-rich linker. Within the flavin domain, a putative cellulose-binding domain (CBD) was found by alignment to the hypothesized CBD of P. chrysosporium CDH. The CBD of CDH appeared to be structurally unrelated to other CBDs which have been reported. / A cDNA clone encoding T. versicolor CDH was isolated by RT-PCR. Using this clone, three vectors for the heterologous expression in Aspergillus oryzae of CDH were prepared. These vectors were built by performing in-frame fusions of the cDNA to control sequences from the highly expressed A. oryzae amylase gene. These vectors were transformed into A. oryzae and one strain was isolated which contained the expression construct DNA. / A rapid method for cloning cdh-like genes was developed. Using short stretches of amino acids completely conserved within T. versicolor and P. chrysosporium CDH, PCR primers were designed to amplify a homologous gene from other fungi. The primers were tested using genomic DNA of Pycnoporus cinnabarinus. A 1.8-kb fragment of P. cinnabarinus cdh was thereby amplified and cloned, and its sequence was determined. The three CDHs displayed very high homology at the amino acid level. / Finally, to probe the role of CDH in lignocellulose degradation by T. versicolor, a "knockout" vector was constructed consisting of a phleomycin-resistance cassette inserted into the protein coding sequence of cloned T. versicolor cdh. T. versicolor was transformed with the knockout vector and the transformants were analyzed for their CDH-producing phenotype. Three isolates were found that produced no detectable CDH. Biobleaching and delignification by the CDH(-) strains appeared to be unaffected, suggesting that CDH does not play an important role in these processes.

Lignocellulose can become the best opportunity for the society to reduce its dependency on the harmful petroleum based products as well as to produce clean energy. In each part of the production cycle, biomass based products have a better environmental profiles than their petroleum based counterparts. Woody biomass has a vast availability, but it suffers from recalcitrance that is mostly caused by lignin that is functioning as a matrix, surrounding and binding the carbohydrates that are currently the most valuable of the wood components. Lignin-carbohydrate (LC) bonds are believed to be a key element in this recalcitrance and research has shown that these types of bonds are common in wood. These bonds are important in an economical point of view as well, as e.g. residual lignin structures in pulp (lignins bonded to the cellulose and hemicelluloses) require expensive bleaching sequences for their removal. The LC-structures can also be exploited technically as we now have demonstrated. We developed a method that utilizes phenolic end groups that are bonded to different hemicelluloses for cross-linking. The enzyme laccase was used for the cross-linking to create a cost-efficient processing scheme to both isolate and increase the molecular weight of the hemicelluloses. Membrane filtration was used as the key separation technique, which enables the establishment of industrial scale production. The final product had improved mechanical and thermal properties and could be used e.g. as barrier film component in renewable packaging. Nanocomposite formation with nanofibrillated cellulose was also studied. This improved the film properties further. The complexes are also possible to use as model compounds for lignin-carbohydrate complexes in wood. This technique can also be seen to mimick the lignification and lignin-carbohydrate network formation phenomena in plants enabling the formation of entire networks of wood components. Our results suggests that the side chains of hemicellulose might play an important role in network formation and that hemicellulose molecules can carry more than one lignin phenolic end group to fulfill this capability. / <p>QC 20140825</p>

The conversion of switchgrass lignin, a renewable source for chemicals and fuels, is investigated using reactions such as depolymerization, hydrodeoxygenation and alkylation. First, the lignin is converted into oils containing phenol, substituted guaiacols and other smaller lignin fragments using formic acid and Pt/C through a batch process. A long reaction time was observed to crucial to yield oils with the highest fraction of lower molecular weight compounds with the lowest O/C and highest H/C molar ratio. Second, the zeolite catalyzed gas phase alkylation of phenol, a model compound for the lignin oil, with propylene was investigated. Zeolite pore topology and acid strength were shown to influence the selectivity for the target product, 2-isopropylphenol. This work shows that the conversion of lignin to useful products is possible and suggests some future work to consider before it can be implemented practically.