Dissertationen zum Thema „Fungal fiber“

The aim of this project was to address the food waste problem, particularly bread waste, to some extent by producing monofilaments obtained from wet spinning of fungal hydrogel through the cultivation of Rhizopus delemar on bread waste. The project had two phases. Firstly, the possibility of production of chitosan fiber with wet spinning (using different acids) was evaluated, the process was optimized, and then applied to the production of fungal fiber. Regarding first stage of the project, adipic acid, a non-toxic solvent with two carboxyl groups, was used as acting physical crosslinker between the chitosan chains, resulting in improving properties of the monofilaments. Adipic acid performance was compared with conventional solvents, such as citric, lactic, and acetic acids. By injecting chitosan solutions into a coagulation bath (EtOH or NaOH 1M or EtOH-NaOH or H2SO4-EtOH), monofilaments were formed. Scanning electron microscopy showed that uniform chitosan monofilaments with smooth surface were formed using adipic and lactic acids. In general, fibers obtained from adipic acid displayed higher mechanical strength (Young’s modulus of 4.45 GPa and tensile strength of 147.9 MPa) than that of monofilaments produced using conventional solvents. Fiber dewatering with EtOH before drying led to greater fiber diameter and lower mechanical strength. As the second stage of this study, Rhizopus delemar was cultivated on bread waste in shake flasks and 1.3 M3 bioreactor. While different combinations of ground bread and K2HPO4 was used as the substrate for shake flask cultivations, white bread waste without K2HPO4 was utilized for scaling up the process, mostly due to the Glucosamine (GlcN) and N-acetyl-glucosamine (GlcNAc) content in the fungal cell wall. GlcN and GlcNA content obtained from ground bread was remarkably higher than that of obtained from combinations of ground bread and K2HPO4 as the substrate. Cultivation in 1.3 M3 bioreactor resulted in about 36 kg wet biomass with a mean of 14.88% dry weight, indicating 5.95 g biomass/L. The biomass yield of 0.15 g dry biomass/g dry bread was achieved. Alkali insoluble material (AIM) was obtained by alkali treatment of biomass. Fungal hydrogel was prepared by adding adipic and lactic acid to AIM, followed by grinding treatment. While hydrogels treated with lactic acid showed better spinnability and gelling ability, the one from adipic acid was not uniform to be wet spun. Considering hydrogels treated with lactic acid, the optimum grinding cycle for more spinnable hydrogel was 6 negative cycles, contributing to the fibers with the tensile strength of around 82 MPa. Such fibers showed antibacterial property against Escherichia coli, making them as a good option for suture applications. However, further in vitro and in vivo trials are essential to test the fungal fiber for such applications.

Establishing a commercial, lignocellulose-based, second-generation ethanol process has received several decades of attention by both researchers and industry. However, a fully economically viable process still remains a long-term goal. The main bottleneck to this achievement is the recalcitrance of lignocellulosic feedstocks, although there are several other factors, such as the huge investment required for second-generation ethanol facilities. An intelligent alternative solution discussed in this thesis is an integrated approach using firstgeneration ethanol plants for second-generation processes. Wheat is the major feedstock for first-generation ethanol in Europe; therefore, wheat-based lignocellulose waste, such as wheat straw, bran, and whole stillage fiber (a waste stream from first-generation wheat-based ethanol plants) was the primary focus of the integration model in this thesis. Since the major share of first-generation ethanol plant economics focuses on the animal feed DDGS (Distillers’ dried gains with solubles), the integration of lignocellulose should be designed in order to maintain DDGS quality. An ethanol-producing edible filamentous fungus, Neurospora intermedia, a potential protein source in DDGS, was considered for use as the fermenting microbe. The morphological and physiological aspects of this fungus were studied in the thesis, leading to the first report of fungal pellet development. An alternative approach of using dilute phosphoric acid to pretreat lignocellulose, as it does not negatively affect fungal growth or DDGS quality, was demonstrated in both the laboratory and on a 1m3 pilot scale. Furthermore, the process of hydrolysis of pretreated lignocelluloses and subsequent N. intermedia fermentation on lignocellulose hydrolysate was also optimized in the laboratory and scaled up to 1 m3 using an in-house pilot-scale airlift bioreactor. Fungal fermentation on acid-pretreated and enzyme-hydrolyzed wheat bran, straw and whole stillage fiber resulted in a final ethanol yield of 95%, 94% and 91% of the theoretical maximum based on the glucan content of the substrate, respectively. Integrating the first- and second-generation processes using thin stillage (a waste stream from first-generation wheat-based ethanol plants) enhanced the fungal growth on straw hydrolysate, avoiding the need for supplementing with extra nutrients. Based on the results obtained from this thesis work, a new model for integrated first- and second-generation ethanol using edible filamentous fungi processes that also adds value to animal feed (DDGS) was developed.

A inoculação de forrageiras com fungos lignocelulolíticos é uma alternativa para melhorar a qualidade destas sem adição de substâncias químicas. Este projeto visa melhorar a degradabilidade de seis forrageiras: Brachiaria decumbens cv. Basilisk, Pennisetum purpureum Schum. cv. Napier, Panicum maximum cv. Aruana, Cenchrus ciliares cv. Buffel, bagaço de cana-de-açúcar e cana-de-açúcar picada por meio do cultivo de quatro fungos do gênero Pleurotus. Outrossim, extrato enzimático, nas concentrações de 2, 4 e 6 mL, produzido pelo fungo P. sajor-caju CCB 020 crescido em vinhaça, durante 6º, 12º e 18º dias, foram aplicados às mesmas forrageiras. As avaliações das dietas foram pela técnica de produção de gases, degradabilidade in vitro e análise bromatológica do substrato. As atividades das enzimas Lacase, Peroxidase, Manganês Peroxidase, Endoglucanase, Exoglucanase e Xilanase foram determinadas, por um período de 24 dias. Os resultados foram estatisticamente avaliados, utilizando SAS® (Statistical Analysis System Inst., Cary, North Carolina). Os fungos crescidos nas forrageiras, produziram enzimas hidrolíticas e oxidativas durante o processo de fermentação, que atuaram na transformação das forrageiras. A quantidade de proteínas aumentou significativamente na cana-de-açúcar picada e capim Buffel, inoculados com os fungos P. ostreatus e P. albidus CCB 068. A forrageira Aruana foi a que melhor respondeu ao tratamento com fungo, sendo que com P. sajor-caju, ao 18º dia de incubação, teve a concentração de acetato 1,24 vezes maior, em relação a amostra controle, e as demais concentrações de AGCC, como propionato e butirato, diminuir e a razão C2:C3 aumentou. A cana-de-açúcar incubada com P. albidus CCB 068, ao 18º dia, diminuiu as concentrações de acetato, propionato e butirato e aumentou a razão C2:C3 em 0,86 - 0,58 - 0,71 - 1,48 vezes, respectivamente. O bagaço inoculado ao 24º dia com P. albidus provocou aumento nas concentrações de acetato, propionato e butirato e diminuição na razão C2:C3, em 1,14 - 1,75 - 1,32 - 0,64 vezes, respectivamente, e com P. ostreatus mostrou o mesmo comportamento. Com os demais tratamentos nenhum efeito significativo foi observado, pelo teste de Tukey a 5%, entretanto ocorre tendência de aumentar a razão C2:C3 com o tratamento com os fungos. Na produção de gases e degradabilidade in vitro das forrageiras, utilizando os extratos enzimáticos, ocorreu aumento com a concentração aplicada. Efeitos significativos foram observados para as forrageiras Brachiaria, Napier e Aruana, já as forrageiras Buffel, bagaço de cana-de-açúcar e cana-de-açúcar picada não mostraram diferenças significativas, pelo teste de Tukey a 5%. Diferenças nas análises de AGCC não foram significativas, pelo teste de Tukey a 5%, dentro das concentrações ou período de incubação do fungo, nas seis forrageiras, entretanto há uma interação com as atividades das enzimas e pela razão C2:C3 a maior concentração testada de 6 mL, do extrato com o fungo, incubado durante 12º dias, possam ser recomendadas para obtenção de ganho nutricional das dietas. Conclui-se que tanto os fungos inoculados nas forrageiras, como seus extratos enzimáticos foram capazes de modificar a composição bromatológica original das forrageiras. A degradabilidade, ganho nutricional e sustentabilidade ambiental, podem variar substancialmente para os diferentes tipos de forrageiras, espécie de fungo e tempo de incubação. Os resultados obtidos sugerem que os Pleurotus, é um fungo apropriado para melhorar o valor nutritivo das forragerias como alimento para ruminates, melhorando a composição bromatoligica, mas também aumentando a degradabilidade
The inoculation of forages with lignocellulolytic fungi is an option for improving quality without adding chemical products. This project aims to improve the degradability of six forages: Brachiaria decumbens cv. Basilisk, Pennisetum purpureum Schum. Cv. Napier, Panicum maximum cv. Aruana, Cenchrus ciliares cv. Buffel, sugarcane bagasse and chopped sugarcane by cultivating four fungi of the genus Pleurotus. In addition, enzymatic extract, at concentrations of 2, 4 and 6 mL, produced by P. sajor-caju CCB 020, developed in vinasse, during 6, 12 and 18 days were applied to the same forages. The evaluations of the diets were by in vitro technique of gas production and bromatological analysis of the substrate. The activities of the enzymes Lacase, Peroxidase, Manganese Peroxidase, Endoglucanase, Exoglucanase and Xylanase were determined, for a period of 24 days. The results were statistically evaluated by SAS® analysis (Statistical Analysis System Inst., Cary, North Carolina). The fungi grown in the forages, produced hydrolytic and oxidative enzymes during the fermentation process that worked in forage transformation. The amount of proteins increased significantly in the chopped sugarcane and Buffel, inoculated with P. ostreatus and P. albidus CCB 068 fungi. The Aruana grass was the one that responded better to fungi treatments. With P. sajor-caju at the 18th day of incubation, the concentration of acetate was 1.24 times higher than the control sample, and the concentrations of other short chain fatty acids (SCFA), such as propionate and butyrate, tended to decrease and the C2:C3 ratio increased. Sugarcane incubated with P. albidus CCB068 at day 18 decreased acetate, propionate and butyrate concentrations and increased the C2:C3 ratio in 0.86, 0.58, and 0.71 - 1.48 fold, respectively. The bagasse inoculated at the 24th day with P. albidus caused an increase in acetate, propionate and butyrate concentrations and a decrease in the C2:C3 ratio in 1.14, 1.75, and 1.32 - 0.64 fold, respectively, and with P. ostreatus showed the same behavior. With the other treatments, no significant effect was observed by the Tukey test. However, there is a tendency to increase the C2:C3 ratio with the fungus treatment. In the production of gases and in vitro degradability of forages, using the enzymatic extracts, an increase with the applied concentrations occurred. Significant effects were observed for the Brachiaria forage at 5%, while forage Buffel, sugarcane bagasse and chopped sugarcane did not show significant differences by Tukey\'s test at 5%. Differences in the SCFA analyzes were not significant, by the Tukey\'s test at 5%, within the concentrations or incubation period of the fungus, in the six forages studied. However there is an interaction with the activities of the enzymes and the C2:C3, the highest concentration tested (6 mL) of the fungi extracts, incubated for 12 days, can be recommended to obtain increase in nutritional value of the diets. It is concluded that both the fungi inoculated on the forage and their enzymatic extracts were able to modify the original bromatological composition of the forage. The degradability, increase of nutritional value and environmental sustainability, can vary substantially for different types of forages, species of fungus and incubation time. The results suggest that Pleurotus is an appropriate fungus for improving the nutritive value of forage crops as feed for ruminates, improving the bromatoligic composition, but also enhancing the degradability

The rumen microbiome constitutes a unique genetic resource of plant fiber degrading microbial enzymes that could be used for agricultural and industrial purposes. Anaeromyces mucronatus is a poorly characterized anaerobic lignocellulolytic fungus in the rumen. This thesis aimed at better understanding A. mucronatus YE505 and the particle associated rumen microbiota based on transcriptomic and metatranscriptomic approaches. High quality RNA was isolated from the fiber-associated rumen sample based on an improved RNA extraction method. A transcriptomic study was performed to investigate the expression of the fiber degrading system of A. mucronatus YE505, and the functional diversity of the fiber-associated eukaryotes from the rumen of muskoxen (Ovibos moschatus) was explored by a metatranscriptomic study. Much carbohydrate degradation related protein modules were detected. This study established effective approaches to characterizing the functional contents of rumen eukaryotic microbiome as well as rumen fungi, and identified several candidate genes that merit further investigation.
xiv leaves : ill. (some col.) ; 29 cm

Wood inhabiting fungi oposes a great problem for preservation of wooden surfaces everywhere, being the main problem of economic losses of wooden products.

A reference collection consisting of 9 different genus constituting of 21 different strains of wood-inhabiting fungi was used for identification of unknown species of mould and blue stain fungi on wood. The fungus DNA from the samples was isolated from malt extract agar. PCR (Polymerase Chain Reaction) was conducted on rDNA ITS1 and ITS2 regions for amplification of the DNA. The 21 samples were collected to a reference collection for identification of unknown species of fungi on wooden field samples using PCR and T-RFLP (Terminal Restriction Fragment Length Polymorphism).

PCR-based methods, sequencing and T-RFLP were proven to be simple and

accurate methods for detection and identification of fungi in their early stage.

L'étude de l'impact des champignons sur les processus géologiques qui altèrent les mineraux s'appelle la “géo-mycologie”. Les serpentinites sont des roches de la famille des ophiolites et peuvent contenir du chrysotile (amiante serpentine). La réactivité des fibres d'amiante est due à la composition chimique de surface et principalement à la présence d'ions métalliques qui peuvent catalyser des réactions chimiques dangereuses. L'amiante est un problème ambiantale à cause de la présence de roches contenant ce minéral mais aussi à cause des anciennes mines d'amiante. La revalorisation (remediation) de ces sites naturellement contaminés passe par la modification de la toxicité des fibres. Les champignons sont des bons candidats pour la bioremediation de l'amiante. L'objectif de cette thèse est l'isolement de souches fongiques à partir de sols serpentiniques afin de savoir quelles sont les espèces fongiques les plus abondantes dans ces sols et de sélectionner les souches les plus efficaces pour leur interaction avec les fibres d'amiantes. Les modifications des fibres et l'altération du métabolisme fongique ont été considérés. Verticillium leptobactrum semble être l'espèce fongique dominante dans tous les sols serpentiniques examinés : cette espèce n'avait jusque là été que rarement isolée, ce qui rend ces résultats intéressants. Trois espèces fongique peuvent extraire Fe et Mg des fibres de amiante (chrysotile et crocidolite), en modifiant la composition et la réactivité chimique de la surface de fibres et la génotoxicité mesuré dans un système acellulaire. Les champignons expriment, en présence des fibres, des enzymes liés à la réponse aux stress oxydants
The interaction of soil fungi with rocks and minerals is called geomycology. Serpentine rocks belong to the ophiolites family and can contain chrysotile (serpentine asbestos). Asbestos fibres reactivity is related to their surface chemical composition, i. G. The presence of iron, catalysing free radicals release, which is harmful for cells and tissues. Asbestos represents an environmental issue, related not only to serpentine rocks naturally exposed and weathered, but also (and mainly) to asbestos mines and wastes. Soil fungi are good candidates for the bioremediation of asbestos rich soils. The main goal of this thesis is the isolation of soil fungi from asbestos rich soils and the selection of the more suitable to interact and modify asbestos fibres in vitro. The metabolic responses of fungi are also investigated. Verticillium leptobactrum is the most abundant specie in all the serpentinic soils considered. This and interesting result, since this specie has bees previously seldom isolated. V. Leptobactrum and other fungal species are able to extract iron and magnesium from chrysotile and crocidolite fibres, modifying their surface chemical composition and reactivity, and their génotoxicity (in acellular experiments). The fungi express anti-oxydant enzymes

Phlebia radiata est un basidiomycete du groupe des champignons de la pourriture blanche, seuls microorganismes connus capables de degrader totalement la lignine du bois. A l'aide de la microscopie electronique a transmission associee a des techniques cytochimiques, nous avons pu mettre en evidence differents modes de degradation provoques par le champignon au cours de l'attaque d'echantillons de bouleau et de peuplier: il peut soit attaquer selectivement la lignine, en provoquant un amincissement progressif des parois secondaires des fibres et / ou en degradant les lamelles mitoyennes, soit degrader simultanement tous les constituants du bois, en perforant les parois et / ou en degradant specifiquement la couche s1 de la paroi. Une etude originale utilisant des anticorps diriges contre des lignines synthetiques nous a permis de visualiser la distribution heterogene des lignines au sein des differentes couches des parois cellulaires. Il apparait que la nature de la lignine a une influence sur le type de degradation. P. Radiata produit des enzymes ligninolytiques, lignine-peroxydases, manganese-peroxydases et laccases, que nous avons localisees au cours de la degradation du bois, grace a des marquages immunocytochimiques. Afin de suivre les enzymes ligninolytiques a un stade tres precoce de leur formation, une approche en biologie moleculaire utilisant des sondes arn a ete engagee pour localiser les arnm codant pour une lignine-peroxydase et pour une laccase de p. Radiata. Ce champignon, comme les autres champignons de la pourriture blanche, presente, par sa capacite a delignifier le bois, un interet potentiel pour l'industrie papetiere. Nous avons montre que la mnp isolee provoque une defibrillation de pates kraft ecrues. Par ailleurs, l'ion manganese complexe a un acide organique, agit egalement en defibrillant les pates. Il apparait que le complexe mniii-oxalate est plus efficace que le complexe mniii-pyrophosphate a blanchir la pate kraft

Koon Chi Man.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2000.
Includes bibliographical references (leaves 158-174).
Abstracts in English and Chinese.
Acknowledgment --- p.i
Abbreviations --- p.ii
Abstract --- p.v
Chinese Abstract --- p.viii
Table of Content --- p.x
Chapter Chapter one: --- Introduction --- p.1
Chapter 1.1 --- Introduction --- p.1
Chapter 1.2 --- Classification of Plant Polysaccharides --- p.2
Chapter 1.2.1 --- Definition of Dietary Fiber --- p.3
Chapter 1.2.2 --- Types of Soluble Dietary Fiber --- p.3
Chapter 1.3 --- Physiological Effect of Fiber --- p.6
Chapter 1.3.1 --- Reduction in Absorption by Viscous Polysaccharides --- p.7
Chapter 1.3.2 --- Gastric Emptying --- p.7
Chapter 1.3.3 --- Effect of Viscous Polysaccharides on Intraluminal Mixing --- p.8
Chapter 1.3.4 --- Effect of Luminal Secretions on Viscosity --- p.9
Chapter 1.4 --- Physicochemical Qualities and Hypocholesterolemic Effects --- p.9
Chapter 1.5 --- Gastrointestinal Events and Hypocholesterolemic Effects --- p.11
Chapter 1.5.1 --- Mouth --- p.11
Chapter 1.5.2 --- Stomach --- p.12
Chapter 1.5.3 --- Small intestine --- p.12
Chapter 1.5.4 --- Large intestine --- p.13
Chapter 1.6 --- Proposed Mechanisms for Hypocholesterolemic Effects --- p.13
Chapter 1.6.1 --- Altered Bile Acid Absorption and Metabolism --- p.14
Chapter 1.6.2 --- Modified Lipid Absorption and Metabolism --- p.15
Chapter 1.6.3 --- Effects of SCFA on Lipid Metabolism --- p.15
Chapter 1.6.4 --- Changed Hormone Concentrations --- p.16
Chapter Chapter Two: --- Materials and Methods --- p.17
Chapter 2.1 --- Materials --- p.17
Chapter 2.1.1 --- Fungus --- p.17
Chapter 2.1.2 --- Animals --- p.17
Chapter 2.1.2.1 --- Golden Syrian Hamster --- p.17
Chapter 2.1.2.2 --- Rabbit --- p.18
Chapter 2.1.3 --- Characterization of Auricularia Polytricha --- p.18
Chapter 2.1.4 --- Chromatographic materials --- p.22
Chapter 2.1.5 --- "Determination of Plasma TC，HDL-C, LDL-C，TG，AST and ALT" --- p.24
Chapter 2.1.6 --- HMG-CoA Reductase Activity Assay --- p.26
Chapter 2.1.7 --- "Quantitative Determination of Liver Cholesterol, Acidic and Neutral Sterol" --- p.27
Chapter 2.1.8 --- Animal Diets --- p.29
Chapter 2.1.8.1 --- Hamster Diets --- p.29
Chapter 2.1.8.2 --- Rabbit Diets --- p.29
Chapter 2.2 --- Methods --- p.33
Chapter 2.2.1. --- Extraction of Water-Soluble AP Polysaccharide (APP) --- p.33
Chapter 2.2.2. --- Characterization of Auricularia Polytricha --- p.34
Chapter 2.2.2.1 --- Determination of carbohydrate content of AP Polysaccharide --- p.34
Chapter 2.2.2.2 --- Determination of uronic acid content of AP Polysaccharide --- p.34
Chapter 2.2.2.3 --- Determination of protein content of AP Polysaccharide by BCA protein assay --- p.35
Chapter 2.2.2.4 --- Determination of component sugar units of AP Polysaccharide --- p.35
Chapter 2.2.2.5 --- Fractionation of AP Polysaccharide --- p.36
Chapter 2.2.2.6 --- Determination of monosaccharides of AP Polysaccharide by HPLC --- p.37
Chapter 2.2.3 --- "Determination of plasma TC, HDL-C, LDL-C,TG,AST and ALT" --- p.39
Chapter 2.2.3.1 --- Plasma Total Cholesterol --- p.39
Chapter 2.2.3.2 --- Plasma HDL-Cholesterol --- p.40
Chapter 2.2.3.3 --- Plasma LDL-Cholesterol --- p.40
Chapter 2.2.3.4 --- Plasma Triglyceride --- p.41
Chapter 2.2.3.5 --- Plasma Aspartate Aminotransferase --- p.41
Chapter 2.2.3.6 --- Plasma Alanine Aminotransferase --- p.42
Chapter 2.2.4 --- HMG-CoA Reductase Activity Assay --- p.42
Chapter 2.2.4.1 --- Preparation of Hepatic Microsome --- p.42
Chapter 2.2.4.2 --- HMG-CoA Activity Assay --- p.43
Chapter 2.2.5 --- Quantitative Determination of Liver Cholesterol --- p.44
Chapter 2.2.5.1 --- Cholesterol Extraction and its Silylation --- p.44
Chapter 2.2.5.2 --- GLC Analysis of TMS-Ether Derivative of Cholesterol --- p.45
Chapter 2.2.6 --- Quantitative Determination of Neutral and Acidic Sterols --- p.45
Chapter 2.2.6.1 --- Separation of Neutral and Acidic Sterols --- p.45
Chapter 2.2.6.2 --- Conversion of Neutral Sterols to its TMS-Ether Derivative --- p.46
Chapter 2.2.6.3 --- Conversion of Acidic Sterols to its TMS-Ether Derivatives --- p.46
Chapter 2.2.6.4 --- GLC Analysis of Neutral and Acidic Sterols --- p.47
Chapter 2.2.7 --- Study of Atherosclerosis of Rabbit --- p.48
Chapter 2.2.7.1 --- Sudan III staining of the thoracic aorta --- p.48
Chapter 2.2.7.2 --- Measurement of atheroma formation in the aorta --- p.49
Chapter 2.2.8 --- Animal Experiments --- p.51
Chapter 2.2.8.1 --- Protective Effect of APP in Hyperlipidemic Study (Exp. 1) --- p.51
Chapter 2.2.8.2 --- Therapeutic Effect of APP in Hyperlipidemic Study (Exp. 2) --- p.52
Chapter 2.2.8.3 --- Dose Response of APP in Hyperlipidemic Study (Exp. 3) --- p.52
Chapter 2.2.8.4 --- Hypolipidemic Effect of Short Chain Fatty Acid (Exp. 4) --- p.53
Chapter 2.2.8.5 --- Effect of APP and SCFA on HMG-CoA Reductase Activity (Exp5） --- p.53
Chapter 2.2.8.6 --- Hypolipidemic and Anti-atherosclerotic Effect of APP (Exp. 6) ´Ø… --- p.54
Chapter 2.3 --- Statistical analysis --- p.54
Chapter Chapter Three: --- Fractionation and Characterization of Auricularia Polytricha Polysaccharide --- p.55
Chapter 3.1 --- Introduction --- p.55
Chapter 3.2 --- Fungal polysaccharides from Auricularia Polytricha --- p.55
Chapter 3.3 --- Results --- p.57
Chapter 3.3.1 --- Extraction and Fractionation of Auricularia Polytricha --- p.57
Chapter 3.3.2 --- Determination of Carbohydrates Content --- p.58
Chapter 3.3.3 --- Determination of Protein Content --- p.61
Chapter 3.3.4 --- Determination of Uronic Acid Content --- p.61
Chapter 3.3.5 --- Determination of component sugars of AP Polysaccharide --- p.65
Chapter 3.3.6 --- Fractionation of AP Polysaccharide --- p.67
Chapter 3.3.7 --- Determination of monosaccharide components of AP Polysaccharide by HPLC --- p.72
Chapter 3.4 --- Discussion --- p.79
Chapter Chapter Four: --- "Protective, Therapeutic and Dose Effect of Auricularia Polytricha Polysaccharide (APP) on Hyperlipidemia" --- p.83
Chapter 4.1 --- Introduction --- p.83
Chapter 4.2 --- Results (Exp. 1) --- p.86
Chapter 4.2.1 --- Body Weight and Food Intake --- p.86
Chapter 4.2.2 --- Effect of APP Supplementation on Hepatic Cholesterol --- p.86
Chapter 4.2.3 --- "Effect of APP Supplementation on Plasma TC, HDL-C and TG" --- p.87
Chapter 4.2.4 --- Effect of APP Supplementation on Fecal Output of Neutral Sterols --- p.94
Chapter 4.2.5 --- Effect of APP Supplementation on Fecal Output of Acidic Sterols --- p.94
Chapter 4.3 --- Discussion (Exp. 1) --- p.99
Chapter 4.4 --- Results (Exp. 2) --- p.102
Chapter 4.4.1 --- Body Weight and Food Intake --- p.102
Chapter 4.4.2 --- Effect of APP Supplementation on Hepatic Cholesterol --- p.102
Chapter 4.4.3 --- Effect of APP Supplementation on Plasma TC and TG --- p.103
Chapter 4.4.4 --- Effect of APP Supplementation on Plasma HDL-C and LDL-C --- p.104
Chapter 4.5 --- Discussion (Exp. 2) --- p.109
Chapter 4.6 --- Results (Exp. 3) --- p.111
Chapter 4.6.1 --- Body Weight and Food Intake --- p.111
Chapter 4.6.2 --- Dose Response of APP Supplementation on Hepatic Cholesterol --- p.111
Chapter 4.6.3 --- Dose Response of APP Supplementation on Plasma TG --- p.112
Chapter 4.6.4 --- Dose Response of APP Supplementation on Plasma HDL-C and LDL-C --- p.112
Chapter 4.6.5 --- Dose Response of APP Supplementation on ALT and AST Activity --- p.113
Chapter 4.6.6 --- Dose Response of APP Supplementation on Fecal Output of Neutral and Acidic Sterols --- p.113
Chapter 4.7 --- Discussion --- p.121
Chapter Chapter Five: --- Hypolipidemic Effect of Short Chain Fatty Acids --- p.123
Chapter 5.1 --- "Introduction (Exp. 4,5)" --- p.123
Chapter 5.2 --- "Results (Exp. 4,5)" --- p.125
Chapter 5.2.1 --- Body Weight and Food Intake --- p.125
Chapter 5.2.2 --- Effect of SCFA Supplementation on Hepatic Cholesterol --- p.125
Chapter 5.2.3 --- "Effect of SCFA Supplementation on Plasma TG, HDL-C and LDL-C" --- p.128
Chapter 5.2.4 --- Effect of SCFA Supplementation on AST and ALT Activity --- p.128
Chapter 5.2.5 --- Effect of SCFA supplementation on HMG-CoA Reductase Activity --- p.133
Chapter 5.3 --- "Discussion (Exp. 4,5)" --- p.135
Chapter Chapter Six: --- Hypolipidemic and Antiatherosclerotic Effect of APP --- p.137
Chapter 6.1 --- Introduction (Exp. 6) --- p.137
Chapter 6.2 --- Results (Exp. 6) --- p.139
Chapter 6.2.1 --- Body Weight and Food Intake --- p.139
Chapter 6.2.2 --- Effect of APP Supplementation on Hepatic Cholesterol --- p.139
Chapter 6.2.3 --- "Effect of APP Supplementation on Plasma TG, HDL- and LDL-C" --- p.141
Chapter 6.2.3 --- Effect of APP Supplementation on AST and ALT Activity --- p.142
Chapter 6.2.5 --- Effect of APP supplementation on HMG-CoA Reductase Activity --- p.146
Chapter 6.2.6 --- Effect of APP supplementation on the Formation of Atheroma --- p.146
Chapter 6.3 --- Discussion (Exp. 6) --- p.151
Chapter Chapter Seven: --- General Discussion and Future Perspectives --- p.153
References --- p.158

Sit Ling.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2001.
Includes bibliographical references (leaves 135-150).
Abstracts in English and Chinese.
Acknowledgment --- p.i
Abbreviations --- p.ii
Abstract --- p.v
Chinese Abstract --- p.vii
Table of Content --- p.ix
Chapter Chapter one: --- General Introduction --- p.1
Chapter 1.1 --- Introduction --- p.1
Chapter 1.2 --- Definition of Dietary Fiber --- p.1
Chapter 1.3 --- Classification of Dietary Fiber --- p.2
Chapter 1.4 --- Hypocholesterolemic Effects of Soluble Dietary Fibers --- p.3
Chapter 1.5 --- Proposed Mechanisms for Hypocholesterolemic Effects --- p.4
Chapter 1.5.1 --- Alter Eating Pattern --- p.4
Chapter 1.5.2 --- Delay Gastric Emptying --- p.4
Chapter 1.5.3 --- Modify Lipid Digestion and Absorption --- p.5
Chapter 1.5.4 --- Effects of SCFA on Lipid Metabolism --- p.6
Chapter 1.5.5 --- Enhance Bile Acid Excretion --- p.7
Chapter 1.6 --- Auricularia polytricha --- p.8
Chapter Chapter Two: --- Chemical Analysis of Auricularia polytrica --- p.11
Chapter 2.1 --- Introduction --- p.11
Chapter 2.2 --- Materials and Methods --- p.12
Chapter 2.2.1 --- Extraction and Fractionation of Auricularia polytricha --- p.12
Chapter 2.2.2 --- Determination of Carbohydrate Content --- p.12
Chapter 2.2.3 --- Determination of Protein Content --- p.13
Chapter 2.2.4 --- Determination of Uronic Acid Content --- p.13
Chapter 2.2.5 --- Determination of Molecular Weight by Gel Filtration Chromatography --- p.14
Chapter 2.2.6 --- Determination of Monosaccharide Components by HPLC --- p.15
Chapter 2.3 --- Results --- p.18
Chapter 2.3.1 --- Yield of Auricularia polytricha polysaccharides --- p.18
Chapter 2.3.2 --- Carbohydrate Content of APPs --- p.18
Chapter 2.3.3 --- Protein Content of APPs --- p.18
Chapter 2.3.4 --- Uronic Acid Content of APPs --- p.19
Chapter 2.3.5 --- Molecular Weight of APPs --- p.22
Chapter 2.3.6 --- Monosaccharide Components of APPs --- p.27
Chapter 2.4 --- Discussion --- p.33
Chapter Chapter Three: --- Hypolipidemic Effects of APPs --- p.36
Chapter 3.1 --- Introduction --- p.36
Chapter 3.2 --- Materials and Methods --- p.38
Chapter 3.2.1 --- Golden Syrian Hamster --- p.38
Chapter 3.2.2 --- Animal Experiments --- p.40
Chapter 3.2.2.1 --- Protective Effect and Dose Response of APPs (Exp. 1) --- p.40
Chapter 3.2.2.2 --- Therapeutic Effect of APPs (High-cholesterol Diet) (Exp. 2) --- p.40
Chapter 3.2.2.3 --- Therapeutic Effect of APPII (Normal Diet) (Exp. 3) --- p.41
Chapter 3.2.2.4 --- Effect of APPs on HMG-CoA Reductase and AC AT Activity (Exp. 4) --- p.42
Chapter 3.2.3 --- Determination of Plasma AST and ALT --- p.42
Chapter 3.2.4 --- "Determination of Plasma TC, LDL-C, HDL-C and TG" --- p.43
Chapter 3.2.5 --- Quantitative Determination of Hepatic and Heart Cholesterol --- p.43
Chapter 3.2.6 --- Quantitative Determination of Perirenal Adipose Tissue Triglyceride --- p.44
Chapter 3.2.7 --- Statistical analysis --- p.45
Chapter 3.3 --- Results (Exp. 1) --- p.47
Chapter 3.3.1 --- Food Intake and Growth --- p.47
Chapter 3.3.2 --- Effect of APPs on Plasma AST and ALT --- p.47
Chapter 3.3.3 --- "Effect of APPs on Plasma TC, LDL-C, HDL-C and TG" --- p.53
Chapter 3.3.4 --- Effect of APPs on Hepatic and Heart Cholesterol --- p.59
Chapter 3.4 --- Discussion (Exp. 1) --- p.64
Chapter 3.5 --- Results (Exp. 2) --- p.67
Chapter 3.5.1 --- Food Intake and Growth --- p.67
Chapter 3.5.2 --- Effect of APPs on Plasma AST and ALT --- p.67
Chapter 3.5.3 --- "Effect of APPs on Plasma TC, LDL-C, HDL-C and TG" --- p.67
Chapter 3.5.4 --- Effect of APPs on Hepatic and Heart Cholesterol --- p.71
Chapter 3.6 --- Discussion (Exp. 2) --- p.74
Chapter 3.7 --- Results (Exp. 3) --- p.76
Chapter 3.7.1 --- Food Intake and Growth --- p.76
Chapter 3.3.2 --- Effect of APPII on Plasma AST and ALT --- p.76
Chapter 3.7.3 --- "Effect of APPII on Plasma TC, LDL-C, HDL-C and TG" --- p.76
Chapter 3.7.4 --- Effect of APPII on Hepatic and Heart Cholesterol --- p.80
Chapter 3.8 --- Discussion (Exp. 3) --- p.83
Chapter Chapter Four: --- Influences of APPs on Cholesterol Homeostasis --- p.84
Chapter 4.1 --- Introduction --- p.84
Chapter 4.2. --- Materials and Methods --- p.87
Chapter 4.2.1 --- HMG-CoA Reductase Activity Assay --- p.87
Chapter 4.2.1.1 --- Preparation of Hepatic Microsome --- p.87
Chapter 4.2.1.2 --- HMG-CoA Reductase Activity Assay --- p.87
Chapter 4.2.2 --- ACAT Activity Assay --- p.88
Chapter 4.2.2.1 --- Preparation of Hepatic and Intestinal Microsome --- p.89
Chapter 4.2.2.2 --- ACAT Activity Assay --- p.89
Chapter 4.2.3 --- Quantitative Determination of Neutral and Acidic Sterols --- p.90
Chapter 4.2.3.1 --- Extraction of Neutral and Acidic Sterols --- p.90
Chapter 4.2.3.2 --- Conversion of Neutral Sterols to its TMS-Ether Derivative --- p.91
Chapter 4.2.3.3 --- Conversion of Acidic Sterols to its TMS-Ether Derivatives --- p.91
Chapter 4.2.3.4 --- GLC Analysis of Neutral and Acidic Sterols --- p.92
Chapter 4.3 --- Statistic Analysis --- p.93
Chapter 4.4 --- Results (Exp. 4) --- p.94
Chapter 4.4.1 --- Effect of APPs on Hepatic HMG-CoA Reductase Activity --- p.94
Chapter 4.4.2 --- Effect of APPs on Hepatic and Intestinal AC AT Activity --- p.94
Chapter 4.4.3 --- Effect of APPs on Fecal Excretion (Exp. 1 & 4) --- p.98
Chapter 4.5 --- Discussion (Exp. 4) --- p.105
Chapter Chapter Five: --- Hypolipidemic and Antiatherosclerotic Effect of APPII in Rabbit --- p.110
Chapter 5.1 --- Introduction --- p.110
Chapter 5.2 --- Materials and Methods --- p.113
Chapter 5.2.1 --- New Zealant White Rabbit --- p.113
Chapter 5.2.2 --- Hypolipidemic and Anitatherosclerosis Effect of APPII (Exp. 5) --- p.113
Chapter 5.2.3 --- Measurement of Atheroma Formation --- p.115
Chapter 5.3 --- Results (Exp. 5) --- p.117
Chapter 5.3.1 --- Food Intake and Growth --- p.117
Chapter 5.3.2 --- Effect of APPII on Plasma AST and ALT --- p.117
Chapter 5.3.3 --- "Effect of APPII on Plasma TC, LDL-C, HDL-C and TG" --- p.117
Chapter 5.3.4 --- Effect of APPII on Hepatic and Heart Cholesterol --- p.125
Chapter 5.3.5 --- Effect of APPII on Perirenal Adipose Tissue Triglycerige Composition --- p.125
Chapter 5.3.6 --- Effect of APPII on the Formation of Atheroma --- p.125
Chapter 5.4 --- Discussion (Exp. 5) --- p.130
Chapter Chapter Six: --- Conclusion --- p.132
References --- p.135

Growing economic and environmental concerns have resulted in a move to reduce fresh water utilization at mechanical pulp and paper mills. This involves retaining and reusing process waters, which consequently provides benefits such as lower effluent treatment costs, energy savings, and improved environmental performance. However, mills experience major problems when attempting water system closure, such as the accumulation of dissolved and colloidal substances (DCS) within the water system. The buildup of DCS can lower paper quality, increase rates of corrosion, and reduce paper machine runnability. Consequently, efficient and cost effective treatment strategies are required for the removal of these contaminants in order for mills to achieve effective water system closure. Unfortunately, existing treatment technologies are far from ideal, and as a result, fresh water usage at mills remains high. Our group has previously demonstrated the potential of a fungal and enzyme treatment strategy for the removal of DCS from mill process waters. To further the development of this treatment strategy, the work conducted in this thesis initially focused on fungal growth and enzyme production, followed by an evaluation of enzyme thermostability. Fungal growth was then carried out in a larger scale (18 L) bioreactor to identify potential changes to extracellular enzyme production as a result of scale-up. The enzymes produced in both cases were used to treat fresh white water and pulp from a mechanical pulp and paper mill, and changes to white water, fiber, and paper properties were determined. The white-rot fungus, Trametes versicolor, utilized in this treatment strategy did not grow well at 45 or 60°C, and the production of cellulolytic, hemicellulolytic, lipolytic, and oxidative enzymes were significantly reduced when compared to the values detected after growth at 30°C. However, these same enzymes were found to maintain substantial percentages of their original activity when incubated at 65°C for extended periods of time. The scale-up of fungal growth in a bioreactor produced very comparable enzyme activities to those determined previously in shake flask cultures. Enzymatic treatment of fresh white water and pulp resulted in changes to both white water and fiber properties. The average colloidal particle size within the treated white water was reduced when compared to untreated white water, while the average molecular weight of the phenolic compounds present in the white water increased. Additionally, the average zeta potential of the colloidal particles was decreased in the treated water, indicating reduced colloidal particle stability. The changes made to the white water contaminants as a result of the enzymatic treatments significantly enhanced DCS removal by precipitation when an alum post treatment step was employed. Mechanical pulp added to the enzyme treated white water showed increased surface charge when compared to pulp blended with the corresponding control water. Handsheets were prepared from enzyme treated or control pulps and were formed in enzyme treated or control white waters, with or without alum post treatment. Paper consolidation and dry strengths were unaffected by any of the treatments, as was illustrated by the very similar densities, scattering coefficients, and tensile, tear, burst, and zero-span indices measured. Enzyme treatment of white water and pulp, followed by alum post treatment significantly enhanced paper surface properties by lowering handsheet roughness and porosity. However, enzyme addition hindered paper optical properties, resulting in lower brightness values. The loss in pulp brightness was largely overcome after a twostage hydrogen peroxide brightening sequence was applied. Brightening resulted in the enzyme treated pulp reaching a comparable final brightness to that of bleached control pulp.

碩士
國立臺南大學
自然科學教育學系碩士班
93
The aim of the study was to characterize the degradation activity of coprophilous fungi on three types domestic forage samples. Growth curves of nine fungi on nilegrass No1. (Acroceras macrum Stapf AC15), napiergrass taishigrass No.2 (Pennisetum purpureum Schum A146 × Pennisetum purpureum Schum A149) and pangolagrass (Digitaria decumbens Stent A254) forage are demonstrated. After initial experiments for testing the potential of cellulase of coprophilous fungi were carried out by the interaction of the directed Congo red on cellulose based Mandels-Reese agar at different pH, Circinella mucoroides, Ascodesmis nigricans, Ascobolus scatigenus, Coprinus patouillardii and Coprinus stercoreus were selected to estimate the digested effect on forage fiber by series detergent methods including neutral and acid detergent fiber (ADF, NDF) and ash content based on the phases of fungal growth pattern. The digestion of cellulose, hemicellulose and lignin were computed by variations among NDF, ADF and ash digestion values. Results demonstrated that the variations and relationship patterns between NDF, ADF and ash varied from sample to sample. While computed on series digestion values, presumably conclusions showed that C. patouillardii and C. stercoreus carry the digestion ability of cellulose, hemicellulose and lignin on nilegrass No1. forage; A. scatigenus and C. stercoreus on napiergrass taishigrass No.2 forage. The usages of soluble CHO for growth of Ascobolus scatigenus were higher than C. mucoroides and A. nigricans on nilegrass No1. forage. C. mucoroides only uses soluble CHO on napiergrass taishigrass No.2 forage. The highest ash content was in degradation process of C. mucoroides and C. patouillardii. However, the ash contents of dry matter can be an influenced factor in the determination of fiber digestion.

Biocatalysts are important tools for harnessing the potential of wood fibres since they can perform specific reactions with low environmental impact. Challenges to bioconversion technologies as applied to wood fibres include low accessibility of plant cell wall polymers and the heterogeneity of plant cell walls, which makes it difficult to predict conversion efficiencies. White-rot fungi are among the most efficient degraders of plant fibre (lignocellulose), capable of degrading cellulose, hemicellulose and lignin. Phanerochaete carnosa is a white-rot fungus that, in contrast to many white-rot fungi that have been studied to date, was isolated almost exclusively from fallen coniferous trees (softwood). While several studies describe the lignocellulolytic activity of the hardwood-degrading, model white-rot fungus Phanerochaete chrysosporium, the lignocellulolytic activity of P. carnosa has not been investigated. An underlying hypothesis of this thesis is that P. carnosa encodes enzymes that are particularly well suited for processing softwood fibre, which is an especially recalcitrant feedstock, though a major resource for Canada. Moreover, given the phylogenetic similarity of P. carnosa and P. chrysosporium, it is anticipated that the identification of pertinent enzymes for softwood degradation can be more easily conducted. In particular, this project describes the characterization of P. carnosa in terms of the growth conditions that support lignocellulolytic activity, the effect of enzymes secreted by P. carnosa on the chemistry of softwood feedstocks, and the characterization of the corresponding secretome using proteomic techniques. Through this study, cultivation methods for P. carnosa were established and biochemical assays for protein activity and quantification were developed. Analytical methods, including FTIR and ToF-SIMS were used to characterize wood samples at advancing stages of decay, and revealed preferential degradation of lignin in the early stages of growth on all softwoods analyzed. Finally, an in depth proteomic analysis of the proteins secreted by P. carnosa on spruce and cellulose established that similar sets of enzyme activities are elicited by P. carnosa grown on different lignocellulosic substrates, albeit to different expression levels.