Dissertations / Theses on the topic 'Ganoderma lucidum'

Ganoderma species are one of the most widely researched fungi because of their reported potent bioactive properties. Although there is much information related to American, European and Asian isolates, little research has been conducted on Australian Ganoderma isolates. Ganoderma may only be imported into Australia under strict quarantine conditions, therefore, the isolation of a native strain that possesses bioactivity may be industrially and commercially significant. Three Australian species of this wood-decomposing fungus were isolated in northern Queensland. In this study, they have been identified as three separate species. Further, they have been studied to determine their optimal growth conditions in liquid culture and assessed for their antibacterial properties. Phylogeny inferred from the Internal Transcribed Spacer Regions (ITS) from the DNA sequences resolved the three Australian Ganoderma species into separate clades. Two isolates were identified to be isolates of Ganoderma cupreum (H1) and Ganoderma weberianum (H2). The third isolate could only be identified to the genus level, Ganoderma species, due to the lack of informative data that could be used for comparison. The effects of short term and long term storage on the viability of the fungi were investigated on agar plates, agar slants and balsa wood at varying temperatures ranging from 10 to 45�C. The most appropriate storage conditions were determined to be �80�C on balsa wood chips for periods of up to 2 years without subculture, and on agar slants at 4�C for up to a maximum of eight weeks. Light was observed to be detrimental to the survival of Ganoderma H1 and Ganoderma H2 during storage. Growth trials using potato dextrose agar plates determined the optimal temperature and pH for mycelial growth to be 30�C and a pH of 6, for all isolates. Subsequent growth trials in liquid media found that glucose, as the carbohydrate source, supported the greatest mycelial growth of Ganoderma H1 and Ganoderma H2 and that galactose and fructose supported the greatest growth of Ganoderma H3. Abstract ii Aqueous (hot water) and organic (hexane (HEX), dichloromethane (DCM), ethyl acetate (EtOAc), methanol (MeOH)) extracts from the liquid cultivated mycelium were assessed for their antibacterial activity using disc diffusion assays. Extracts from the mycelium of Ganoderma H1 exhibited activity against a greater number of Gram positive bacteria than those from Ganoderma H2 and H3. Subsequent studies on the DCM and EtOAc extracts from Ganoderma H1 determined the MIC and MBC against a number of Gram positive bacteria, including Bacillus cereus, B. subtilis, Enterococcus faecalis, Streptococcus pyogenes, Staphylococcus aureus, S. epidermidis and Listeria monocytogenes, as well as Clostridium species, including Clostridium perfringens, C. sporogenes and C. difficile, and some methicillin resistant Staphylococcus aureus (MRSA) strains. Time course growth assays confirmed that the DCM and EtOAc extracts predominantly exhibited bactericidal activity. Finally, the active compounds were determined to be terpenoid in structure with some phenolic groups attached.

Ganoderma lucidum and G. tsugae are two members of the G. lucidum complex. The authenticity of the two wood-rotting species was demonstrated by comparative studies. Ganoderma lucidum is restricted to hardwoods. Its "smooth" walled basidiospores were characterized by narrow, numerous inter-wall pillars. Isolates of G. lucidum produced chlamydospores in culture and had an average growth of 7.8 mm/da at their optimum temperature range of 30-34 C. Ganoderma tsugae is restricted to conifers. Its basidiospores were "rough" walled and had broad inter-wall pillars. Isolates of G. tsugae did not produce chlamydospores in culture and had an average growth of 2.1 mm/da at the optimum temperature range of 20-25 C. Mating systems were determined for both species as heterothallic and tetrapolar. Interspecific matings of homokaryons were incompatible. Homokaryons of a European G. resinaceum isolate were interfertile with homokaryons from North American collections of G. lucidum. The ability of G. lucidum and G. tsugae to decay wood in vitro was studied using the following woods in agar block decay chambers: grape, oak, mesquite, white fir, and Douglas-fir. Grape wood lost the most weight while mesquite the least. G. lucidum isolates generally caused greater weight loss of all woods than did G. tsugae isolates. Both Ganoderma species caused simultaneous decay in all woods. However, chemical analyses of the decayed blocks indicated that selective delignification by both species also occurred in grape and white fir blocks but not in oak or Douglas-fir blocks. Scanning electron microscopy demonstrated various stages of selective delignification and simultaneous decay of all woods tested. Isolates of Ganoderma lucidum infected Dog Ridge variety grape plants, grown in the greenhouse, from below-ground wood block inoculations. Twenty-four plants were inoculated: one plant died and 4 other plants declined. After 24 months reisolations yielded only G. lucidum from the five declining plants, demonstrating pathogenicity. The fungus developed in the heartwood and, in later stages, invaded the sapwood. Infected plants developed water stress symptoms with leaves wilting, yellowing, and dying. Field grape plants inoculated with the fungus developed decay columns as large as 42 cm in 17 mons. Decay was limited to the heartwood; no foliar symptoms occurred.

Ganoderma lucidum spores (GLS) have attracted increasing attention for its versatile biological activities, particularly in cancer therapy. The resilient chitin bilayer of sporoderm is conventionally regarded as an obstacle in the exploitation of bioactive ingredients. Present study found that ethanol extract of broken GLS was able to inhibit cancer cells, however, water extract, especially medium extract (containing serum protein) from unprocessed GLS have also demonstrated anti-proliferative effects on cancer cells. The effectiveness of GLS extract on the inhibition of a series of human cancer cells, namely, osteosarcoma, neuroblastoma, myeloid leukaemia and breast cancer, has been compared, and DNA assays showed that the GLS extract is more efficient in inhibiting neuroblastoma but has less effect on osteosarcoma cell line. To overcome the limitations of the existing processing methods of GLS, the feasibility of sonication as a new way to break GLS has been tested. A series of processing parameters, such as sonication power and duration, have been compared to maximise the breaking efficiency. The preservation of bioactive components of GLS (e.g. polysaccharides and ganoderic acids) from sonication processing was revealed by Fourier Transform Infrared Spectroscopy (FTIR) and High Performance Liquid Chromatography (HPLC) analyses. In vitro study showed that sonication processed GLS were able to inhibit breast cancer cells, at dose and time dependent manner, particularly at low pH (6.5), favourable for cancerous cell growth. The inhibitory efficiency of sonication processed GLS on the growth of breast cancer cells was ranked the highest, compared with that of unprocessed GLS and commercially broken GLS. To preserve further the bioactive ingredients of GLS, broken GLS have been encapsulated with alginate by electrospraying (ES). The size of GLS encapsulated alginate (GLS/A) beads was found to affect the in vitro release profiles of bioactive ingredients of GLS, and can be controlled by varying the processing parameters (e.g. crosslinking time, infuse rates and applied voltage). A series of GLS/A beads with mean sizes ranging from 500 to 2500 µm have been produced by ES and the in vitro release profiles of GLS/A beads in simulated gastrointestinal mediums were found to be related to the pH, bead size and drying methods. In summary, an advanced method combining a customised sonication with ES has been developed by setting up a lab-scale production line from processing to encapsulation of GLS. This may pave the way to produce effective GLS products with desirable natural bioactive components for healthcare.

The aim of the present study was to investigate a range of bioprocess strategies aimed at the achievement of maximum biomass yield in submerged cultivation of the Basidiomycete, Ganoderma lucidum. Although there had been previous studies into cultivation of G, lucidum, these had been almost exclusively centred round maximisation of the medically interesting polysaccharide, EPS. The present work is focused on the development of fermentation strategies to achieve this aim, which was a central interest of the sponsor. Additionally, to investigate the process physiology of these complex cultures to help improve the relatively poor, understanding of the bioprocessing of this Basidiomycete fungus and to understand the influence of process variables during submerged cultivation of G. lucidum on growth, polysaccharide production and substrate consumption.

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O fungo Ganoderma lucidum (Fr.) Krast, também conhecido como Lingzhi ou Reish, é um basidiomiceto pertencente à família Ganodermataceae, muito estudado por suas características medicinais. Imunomodulação e atividade antitumoral são algumas de suas indicações mais importantes à saúde humana, devem-se principalmente à presença de polissacarídeos e triterpenóides, que há séculos são utilizados na China, Japão e Coréia por fins terapêuticos. Com a crescente demanda por pesquisa com uso de Ganoderma lucidum no tratamento de doenças, a otimização da produção comercial tornou-se necessária para atender as demandas de mercado e de pesquisa. Assim, os objetivos do presente trabalho foram: 1. avaliar e comparar a eficiência biológica e o rendimento de 7 linhagens de G. lucidum, 351, 1002, 0901, 0809, 339, 144, 202, cedidas por produtores e pesquisadores ao banco de linhagens do Módulo de Cogumelos FCA/UNESP. Todas cultivadas em substrato à base de serragem de eucalipto; 2. quantificar as concentrações de β-glucanas, fenólicos totais e proteínas nas linhagens estudadas. O substrato de cultivo foi composto de 80% de material volumoso (serragem de eucalipto) e 20% de farelos (trigo e soja) contendo 620g cada saco de cultivo. A quantificação das β-glucanas foi realizada por método enzimático; para a quantificação de fenólicos totais, utilizou-se o método de Folin-Ciocalteau e para proteínas totais o método utilizado foi de Lowry. O cultivo, desde a inoculação do substrato até a colheita dos basidiomas, durou, em média, 65 dias. Em relação a eficiência biológica as linhagens obedeceram a seguinte ordem 351 ≥ 1002...
The fungus Ganoderma lucidum (Fr.) Krast , also known as Lingzhi or Reish, is a basidiomycete belonging to Ganodermataceae family, widely studied for its medicinal characteristics . Immunomodulation and antitumor activity are some of his most important insights to human health, are mainly due to the presence of polysaccharides and triterpenoids , which are used for centuries in China, Japan and Korea for therapeutic purposes. With the growing demand for research use of Ganoderma lucidum in treating diseases , optimization of commercial production became necessary to meet market demands and research . The objectives of this study were: 1 . evaluate and 4 compare the biological efficiency and yield of 7 strains of Ganoderma lucidum , 351 , 1002, 0901 , 0809 , 339 , 144 , 202 , assigned by producers and researchers to bank lines Module Mushrooms FCA / UNESP . All grown on substrate with sawdust ; 2 . quantify the concentrations of β - glucan , total phenolics and proteins in the strains studied . The culture substrate was composed of 80 % of bulk material ( sawdust ) and 20% bran (wheat and soybeans ) containing 620g each bag cultivation . Quantification of β - glucans was performed by enzymatic method ; for quantification of total phenolic used the Folin -Ciocalteau total protein and the Lowry method was used . The cultivation , from inoculation to harvest the substrate of the mushroom lasted , on average, 65 days. Relative biological effectiveness strains obeyed the following order 351 ≥ 1002 ≥ 0901 = 0809 = 339 with 15.00% , 13.10 % , 11.77 % , 11.29 % and 10.87 % . The yield followed the same order of significance , and obtained the following values : 5.95, 4.98 , 4.31 , 4.29 , and 4.18 g kg - 1 strains respectively. The line with the highest concentration of β - glucans was 1002 with 10.50 g/100 g of dry mushroom. The strains with the highest concentration of total phenolics were 1002 and 08 ...

Orientador: Marli Teixeira de Almeida Minhoni
Coorientador: Marli Gerenutti
Banca: Diego Cunha Zied
Banca: Adriana Zanin Kronka
Resumo: O fungo Ganoderma lucidum (Fr.) Krast, também conhecido como Lingzhi ou Reish, é um basidiomiceto pertencente à família Ganodermataceae, muito estudado por suas características medicinais. Imunomodulação e atividade antitumoral são algumas de suas indicações mais importantes à saúde humana, devem-se principalmente à presença de polissacarídeos e triterpenóides, que há séculos são utilizados na China, Japão e Coréia por fins terapêuticos. Com a crescente demanda por pesquisa com uso de Ganoderma lucidum no tratamento de doenças, a otimização da produção comercial tornou-se necessária para atender as demandas de mercado e de pesquisa. Assim, os objetivos do presente trabalho foram: 1. avaliar e comparar a eficiência biológica e o rendimento de 7 linhagens de G. lucidum, 351, 1002, 0901, 0809, 339, 144, 202, cedidas por produtores e pesquisadores ao banco de linhagens do Módulo de Cogumelos FCA/UNESP. Todas cultivadas em substrato à base de serragem de eucalipto; 2. quantificar as concentrações de β-glucanas, fenólicos totais e proteínas nas linhagens estudadas. O substrato de cultivo foi composto de 80% de material volumoso (serragem de eucalipto) e 20% de farelos (trigo e soja) contendo 620g cada saco de cultivo. A quantificação das β-glucanas foi realizada por método enzimático; para a quantificação de fenólicos totais, utilizou-se o método de Folin-Ciocalteau e para proteínas totais o método utilizado foi de Lowry. O cultivo, desde a inoculação do substrato até a colheita dos basidiomas, durou, em média, 65 dias. Em relação a eficiência biológica as linhagens obedeceram a seguinte ordem 351 ≥ 1002 ...
Abstract: The fungus Ganoderma lucidum (Fr.) Krast , also known as Lingzhi or Reish, is a basidiomycete belonging to Ganodermataceae family, widely studied for its medicinal characteristics . Immunomodulation and antitumor activity are some of his most important insights to human health, are mainly due to the presence of polysaccharides and triterpenoids , which are used for centuries in China, Japan and Korea for therapeutic purposes. With the growing demand for research use of Ganoderma lucidum in treating diseases , optimization of commercial production became necessary to meet market demands and research . The objectives of this study were: 1 . evaluate and 4 compare the biological efficiency and yield of 7 strains of Ganoderma lucidum , 351 , 1002, 0901 , 0809 , 339 , 144 , 202 , assigned by producers and researchers to bank lines Module Mushrooms FCA / UNESP . All grown on substrate with sawdust ; 2 . quantify the concentrations of β - glucan , total phenolics and proteins in the strains studied . The culture substrate was composed of 80 % of bulk material ( sawdust ) and 20% bran (wheat and soybeans ) containing 620g each bag cultivation . Quantification of β - glucans was performed by enzymatic method ; for quantification of total phenolic used the Folin -Ciocalteau total protein and the Lowry method was used . The cultivation , from inoculation to harvest the substrate of the mushroom lasted , on average, 65 days. Relative biological effectiveness strains obeyed the following order 351 ≥ 1002 ≥ 0901 = 0809 = 339 with 15.00% , 13.10 % , 11.77 % , 11.29 % and 10.87 % . The yield followed the same order of significance , and obtained the following values : 5.95, 4.98 , 4.31 , 4.29 , and 4.18 g kg - 1 strains respectively. The line with the highest concentration of β - glucans was 1002 with 10.50 g/100 g of dry mushroom. The strains with the highest concentration of total phenolics were 1002 and 08 ...
Mestre

The RBF strategy has successfully produced fungal mycelial biomass and EPS in a very strictly regulated manner at high productivity rates compared to batch fermentation. The problematic lag phase and seed culture preparation were reduced in length; harvesting volume doubled, yield of product increased, and medium consumption was reduced in an RBF relative to batch. 80% broth replacement volume and transition phase were optimised. Dispersed mycelial filaments with ovoid-shaped pellets are the typical morphological characteristics associated with EPS production. N-limiting medium in an unbaffled 2.5-L bioreactor stimulated EPS formation during RBF compared to in baffled condition. The current study has managed to alter the molecule's hydrophobicity thus making it water-soluble as proved by compositional analysis and spectroscopy. The sulphated derivative of native glucan was identified as (1, 3)-(Sb(B-D-glucan. Sulphation was an effective approach to improve antibacterial, antifungal, antiproliferative and immunomodulatory (NO stimulation) activity of the sulphated (1,3)-(Sb(B-D-glucan or GS. GS maybe safe in in vitro trials due to its demonstrated lack of toxicity towards a normal human prostate cell line (PN2TA). GS also showed antimicrobial-antifungal-immunomodulatory activities derived from a single compound. Fungal cells tended to grow well in the porous structure of PUF cubes and the RBF using immobilised fungal cells was an efficient method for production of (Sb(B-glucan with a high yield. This study could be beneficial for other medicinal mushroom fermentation.

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Universidade Federal de Pernambuco
Ganoderma lucidum é um basidiomiceto muito popular na medicina tradicional chinesa, sendo chamado de Lingzhi ou cogumelo da imortalidade . Estudos revelaram diversas propriedades farmacológicas benéficas deste fungo, sugerindo sua utilização como auxiliar em diversos tratamentos contra doenças humanas. Por não ser muito comum na natureza, o cultivo é a melhor maneira de suprir um mercado cada vez mais amplo. Dentre os vários métodos de cultivo existentes para a fungicultura, a técnica Jun-Cao é considerada a mais promissora para a produção de cogumelos em escala que atenda à demanda crescente. Sabendo que a bioquímica de um organismo pode variar entre diferentes linhagens, e entendendo a necessidade de descobrir novas variedades de G. lucidum, para ampliar as opções de cultivo deste fungo, este trabalho buscou comparar linhagens comerciais chinesas com linhagens brasileiras inexploradas de G. lucidum, para avaliar o valor bioquimico destas. Os resultados mostram que o cultivo combinado de madeira com gramínea, na técnica Jun-Cao, foi mais promissor no desenvolvimento das linhagens tanto em redução no tempo de cultivo quanto na qualidade e quantidade dos cogumelos. A linhagem brasileira (CC-144) destacou-se tanto na velocidade de crescimento, quanto na produtividade de basidiomas e na concentração bioquímica de proteínas, carboidratos, β- glucanas e fenóis, superando as chinesas e se mostrando uma opção para a fungicultura. A análise RAPD mostrou distinção genética entre as linhagens, descartando a possibilidade de ocorrência de clones

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Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES
Ganoderma lucidum (Curtis) P. Karst is a medicinal fungus (Basidiomycota, Polyporaceae) popularly known as "Lingzhi" in China, "Reishi" in Japan and "Mushroom King" in Brazil, which has been cultivated in several lignocellulosic materials. In the present study, the formation of mycelium as well as the production of ligninolytic enzymes and basidiomata of this species were evaluated in waste fruit shell, leaf and bract endemic palm of the Brazilian semi-arid region, Syagrus coronata (Martius) Beccari (licuri) by solid state fermentation. The optimal cultivation conditions were the same for all three types of substrates tested (pH 6.5, C/N ratio of 40 and 30?C of temperature) and were established from assays in Petri dishes and contour graphic analysis and response surface methodology. The enzymatic activity of laccase and manganese peroxidase were determined using spectrophotometric methods at intervals of 7 days by cultivating the strain in the three substrates selected for 28 days of incubation. The activity of the laccase was determined at 420 nm, using ABTS: 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) as the enzymatic substrate, and the activity of the manganese peroxidase at 610 nm by oxidation of phenol red. The highest peak activity for laccase (13,80 U/L) was found in bract substrate in 14 days of incubation and did not differ significantly from the others at 95% of statistical confidence. That substrate also resulted in the highest peak of activity for manganese peroxidase (14,92 U/L), which only occurred after 28 days of incubation, differing from the previous one at 5% probability. The assay for the production of basidiomata was performed in polypropylene bags. The only substrate that did not promote the formation of the basidiomata was the fruit shell. The biological yield did not differ significantly at 95% confidence in the bract (33,53 g/kg) and leaf (37,48 g/kg) substrates. The biological efficiency also was not statistically different in the bract (3,35%) and leaf (3,75%) substrates. Thus, it is believed that the licuri wastes are potential substrates and can be used in the bioconversion processes for the formation of the mycelium and basidiomata as well for ligninolytic enzymes production in G. lucidum.
Ganoderma lucidum (Curtis) P. Karst ? um fungo medicinal, do Filo Basidiomycota, Fam?lia Polyporaceae, popularmente conhecido como ?Lingzhi? na China, ?Reishi? no Jap?o e ?Cogumelo Rei? no Brasil, que tem sido cultivado em diversos materiais lignocelul?sicos. No presente estudo, foram avaliadas a forma??o de mic?lio, bem como a produ??o de basidiomas e enzimas ligninol?ticas da esp?cie em res?duos de casca de fruto, folha e br?ctea da palmeira end?mica da regi?o semi-?rida brasileira, Syagrus coronata (Martius) Beccari (licuri), por meio de fermenta??o em estado s?lido. As condi??es ?timas de cultivo foram as mesmas para os tr?s tipos de substratos testados (pH 6,5, rela??o C/N de 40 e temperatura de 30?C), sendo estabelecidas a partir do ensaio em placas de Petri e an?lise dos gr?ficos de contorno e superf?cie de resposta. A atividade enzim?tica de lacase e de mangan?s peroxidase foram determinadas empregando m?todos espectrofotom?tricos, em intervalos de 7 dias, atrav?s do cultivo da linhagem nos tr?s substratos selecionados durante 28 dias de incuba??o. A atividade de lacase foi determinada a 420 nm, empregando ABTS: 2,2?-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) como substrato enzim?tico e a de mangan?s peroxidase, a 610 nm, pela oxida??o do vermelho de fenol. O maior pico de atividade para a lacase (13,80 U/L) foi verificado no substrato br?ctea, aos 14 dias de incuba??o, e n?o diferiu significativamente dos demais a 95% de confian?a. Esse substrato tamb?m proporcionou o maior pico de atividade para a mangan?s peroxidase (14,92 U/L), que somente ocorreu aos 28 dias de incuba??o, diferindo estatisticamente dos demais a 5% de probabilidade. O ensaio para a produ??o de basidiomas foi realizado em sacos de polipropileno. O ?nico substrato que n?o promoveu a forma??o de basidiomas foi a casca de fruto de licuri. O rendimento biol?gico n?o diferiu significativamente a 95% de confian?a nos substratos br?ctea (33,53 g/kg) e folha (37,48 g/kg). A efici?ncia biol?gica tamb?m n?o foi estatisticamente diferente nos substratos br?ctea (3,35%) e folha (3,75%). Assim, acredita-se que os res?duos de licuri sejam potenciais substratos e possam ser empregados no processo de bioconvers?o para a forma??o de mic?lio e basidiomas, e tamb?m para a produ??o de enzimas ligninol?ticas de G. lucidum.

碩士
長庚大學
傳統中國醫學研究所
97
Ganoderma lucidum on the "shen nong ben cao jing" (The Divine Husbandmn’s Herbal Foundation Classic) is listed as the top grade of medicine during the time of Chin and Han Dynasty. But over thousand of years, the book has stated a very little about its effects. In the important remedy books such as "qian jin yi fang ", "tai ping sheng hui fang ", "sheng ji zong lu ", "yu yao yuan fang " and "pu yi fang "…etc, it has stated only five prescriptions with the Ganoderma lucidum. The quantity is very rare and makes it difficult for doctors to use it in their daily practice, but this changes to a special appearance in Taoism and local folks as an “immortal elixir” and “propitious pleasant”. In the present days, a total of 76 different types of the Ganoderma have been discovered for medical use. Due to the improving cultivating technique, and over a thousand of researches, they had made the Ganoderma lucidum to play a new role in the health medicine. The old Traditional Chinese Medicine doctors have experienced the rich variety of mushroom species in Taiwan, and re-appraisal of Ganoderma lucidum between the classic and new researches. The clinical experiences among those doctors are our enlightenments to this medical field. We had visited three old Traditional Chinese Medicine doctors, compared and investigated their opinions and thinking process in this research. Dr. Tsai has many years of clinical experiences in the use of Ganoderma lucidum, Fomitopsis pinicolat,Ganoderma applanatum, Hexagona apiaria, Ganoderma sinense, Antrodia cinnamomea, Phellinus igniarius …etc. He has widely used them on patients with liver diseases, insomnia, depression, cardiovascular diseases, renal diseases, cancer, hemorrhoids, constipation…etc. Dr. Cheng has discovered the use of Ganoderma lucidum on patients with chest tightness has a great effect on the illness. However, side effects such as edema and skin rash develop easily on patients. Due to the side effects, Dr. Cheng is no longer using it. Dr. Liu considers that the clinical effects of Ganoderma lucidum are uncertain. Therefore, he does not use any at all. Dr. Tsai only uses the one in the wild. He thinks that by cultivating, the medical effect is not so efficient. He also points out that epiphytic plants will influence the effect of Ganoderma lucidum and this can be used in improvement in cultivating technique and the reference for other doctors. These three doctors have the habits of trying the medication themselves before their use on the patients. They are not only providing a safer medication for the patients, also they are providing ones personal sensitivity to the medication. This can be used as a academic reference to its clinical use.

Relatório de Estágio do Mestrado Integrado em Ciências Farmacêuticas apresentado à Faculdade de Farmácia
Resumo - Relatório de Estágio em Farmácia Comunitária Após a aquisição da formação teórica e prática integrada no Mestrado Integrado emCiências Farmacêuticas, o estudante realiza a unidade de “Estágio Curricular”. Sendo uma das áreas mais relevantes do domínio farmacêutico, a realização de um período de estágio em Farmácia Comunitária é obrigatória.Neste contexto, ingressei na Farmácia Coimbra, localizada no Coimbra Shopping, onde adquiri conhecimentos acerca do funcionamento interno de uma farmácia assim como experienciei o papel do farmacêutico perante a população. O presente relatório será apresentado sob a forma de análise SWOT, por forma a salientar aqueles que, segundo a minha perspetiva, foram os pontos fortes, pontos fracos, oportunidades e ameaças do estágio. Resumo - Relatório de Estágio em Indústria Farmacêutica Como parte integrante do último semestre do Mestrado Integrado em CiênciasFarmacêuticas, os estudantes têm que realizar a unidade de “Estágio Curricular”. Para além da habitual área da Farmácia Comunitária, tive a oportunidade de realizar um estágio numa outra área, mais especificamente em Indústria Farmacêutica, o que me permitiu alargar conhecimentos e explorar vertentes alternativas do domínio farmacêutico.Assim sendo, integrei a equipa da Pharmilab, uma consultora especializada na área doscosméticos, dispositivos médicos, suplementos alimentares e biocidas, onde incorporei oambiente laboratorial, com foco no Controlo da Qualidade. O presente relatório será entãouma análise desta experiência, apresentada sob a forma de análise SWOT, evidenciando ospontos fortes, pontos fracos, oportunidades e ameaças sentidos no decorrer do estágio. Resumo - Potencialidades medicinais de Ganoderma lucidum O Ganoderma lucidum é um basidiomiceto também conhecido como “cogumelo da imortalidade” em países asiáticos, onde a sua utilização remonta a épocas prévias à Era Comum. Este cogumelo tem demonstrado inúmeras potencialidades medicinais atribuídas à elevada diversidade de substâncias ativas presentes no basidiocarpo, micélio e esporos, dentrodos quais se destacam os polissacáridos e os triterpenos. A presente monografia irá abordar quatro destas potencialidades, nomeadamente, as ações imunoestimulantes, antitumorais, antidiabéticas e dislipidémicas.Quanto à estimulação do sistema imunitário salientam-se os polissacáridos, em particularos β-glucanos. Estes têm sido reportados em inúmeros ensaios in vitro e in vivo como capazes de promover a imunidade celular e humoral, assim como de estimular as célulasapresentadoras de antigénios e o sistema mononuclear fagocitário, principalmente através da sua capacidade de induzir a produção de citocinas.A ação antitumoral deste cogumelo deve-se maioritariamente à presença de triterpenos.Apesar da ação imunoestimulante dos polissacáridos ser de elevada relevância no auxílio àterapêutica do indivíduo oncológico, os triterpenos têm demonstrado uma panóplia depropriedades antitumorais, nomeadamente através das suas ações citotóxicas, citostáticas,indução da apoptose e inibição da formação de metástases e da angiogénese.Quanto à ação antidiabética, o G. lucidum tem demonstrado capacidades hipoglicemiantes ehiperinsulinémicas. Enquanto os polissacáridos têm sido reportados como reguladores deenzimas-chave do metabolismo da glucose e indutores da libertação de insulina, os triterpenos exibem capacidade de intervir nas enzimas aldose redutase e α-glicosidase. Existe ainda um proteoglicano que tem revelado propriedades inibitórias da Proteína Tirosina Fosfatase 1B.Relativamente à dislipidémia, o G. lucidum tem manifestado capacidade de promover açõesessencialmente hipocolesterolémicas. Deste modo, os polissacáridos têm sido reportadoscomo capazes de alterar a absorção intestinal do colesterol exógeno, enquanto os esteróis e derivados oxigenados do lanosterol reduzem a síntese hepática do colesterol endógeno.Neste contexto, a presente monografia procede à revisão de literatura sobre os principaisconstituintes do G. lucidum e o seu papel na imunoestimulação bem como no auxílio àterapêutica de algumas das doenças mais relevantes do presente século, por forma a enfatizar as potencialidades medicinais deste cogumelo, assim como averiguar os mecanismos de ação pelos quais atua, e a possível toxicidade e efeitos secundários inerentes à sua utilização.
Abstract - Community Pharmacy Internship Report Following the acquirement of the theoretical and practical training integrated in the Master’s Degree in Pharmaceutical Sciences, students undertake the unit “Curricular Internship”. Being one of the most relevant areas of the pharmaceutical domain, the completion of a probationary period in Community Pharmacy is mandatory.In this regard, I joined Farmácia Coimbra, located at Coimbra Shopping, where I haveknowledge about the internal functioning of a pharmacy as well as experienced thethe pharmacist’s role towards the population. The present report will be displayed under the form of a SWOT analysis, in order to underline those that, according to my perspective, were the strengths, weaknesses, opportunities and threats of the internship. Abstract - Pharmaceutical Industry Internship Report As an integrating part of the last semester of Master’s Degree in Pharmaceutical Sciences, the students have to undertake the unit “Curricular Internship”. In addition to the usual internship in Community Pharmacy, I had the opportunity to take part in another one in a distinct area, more specifically in Pharmaceutical Industry, which allowed me to expand my knowledge and explore alternative paths of the pharmaceutical domain.Therefore, I instated Pharmilab’s team, a specialized consultant in the areas of cosmetics, medical devices, dietary supplements and biocides, where I was incorporated in a laboratory environment, with a focus on Quality Control. The present report will be an assessment of this experience, presented under the form of a SWOT analysis, highlighting the strengths, weaknesses, opportunities and threats perceived throughout the course of the internship. Abstract - Medicinal Potentialities of Ganoderma lucidum Ganoderma lucidum is a basidiomycete mushroom also known as “the mushroom of immortality” in asian countries, where its utilization can be traced back to times prior to the Common Era. This mushroom has demonstrated its countless medicinal potentialities attributed to the high diversity of active substances present in the basidiocarp, mycelia and spores, within which polysaccharides and triterpenes stand out. Herein, four of these potentialities, particularly its immunostimulant, antitumor, antidiabetic and dyslipidaemic actions are approached.In terms of the immune system stimulation polyssacharides stand out, more specifically β-glucans. These have been reported in innumerous in vitro and in vivo studies as capable of promoting cellular and humoral immunity, as well as stimulating antigen presenting cells and phagocytic mononuclear system, mainly through its ability of promoting the production of cytokines.The antitumor activity of this mushroom is predominantly credited to the presence oftriterpenes. Despite the relevance of polyssacharides’ immunostimulating activity in assisting the therapeutics of the oncological patient, triterpenes have shown a panoply of antitumor properties, specifically through cytotoxicity, cytostatic actions, apoptosis induction, and metastasis formation and angiogenesis inhibition.Regarding the antidiabetic action of G. lucidum, hypoglycaemic and hyperinsulinemiccapacities have been demonstrated. While polysaccharides have been reported as modulatorsof key enzymes in the glucose metabolism pathway and as insulin release inducers, triterpenes have evidenced the ability of intervening in the aldose reductase and α-glycosidase enzymes. There is also a proteoglycan that has expressed PTP1B inhibitory properties.Lastly, concerning dyslipidaemia, G. lucidum has displayed the ability to promote essentially hypocholesterolemic actions. For that purpose, polysaccharides have been reported as capable of altering the intestinal absorption of exogenous cholesterol while sterols and oxygenated lanosterol derivatives lower the hepatic synthesis of endogenous cholesterol.Within this context, a literature review upon G. lucidum’s major components and their role in immunostimulation along with the therapeutic aid on some of the most relevant illnesses of the current century shall be carried out, in order to emphasize the medicinal potentialities of this mushroom, as well as ascertain the mechanisms of action by which it operates and the possible toxicity and secondary effects inherent in its use.

碩士
中山醫學大學
營養學研究所
93
Ganoderma lucidum played an important role in Chinese traditional medicine. Triterpenoids and polysaccharides of Ganoderma lucidum were found effective in the treatment of hypertension, chronic bronchitis, allergy, hyperglycemia and cancer. This study was focused on anti-aging effect of Ganoderma lucidum including anti-oxidation ability of plasma, erythrocyte enzyme activities and liver function protection. This was a double blind and cross over study, 39 subjects (21 males and 18 females) involved in this study. The total experimental period was thirteen months. One group of subjects took placebo in the first six months and Ganoderma lucidum in the next six months, one month between this two periods was wash out period. The other group of subjects took Ganoderma lucidum in the first six months and placebo in the next six months. Anthropometric measurements, blood collecting and abdominal ultrasound examination were performed at the initial,3rd,6th,7th,10th,13th months in the testing period. Subjects took 2.7g Ganoderma lucidum extract everyday, equivalent to 1.89g triterpenoids and 1.62g polysaccharides. Results showed that six months continuous intake of Ganoderma lucidum significantly increased total antioxidant capacity, total thiol groups, GSH, reduced TBARS and 8-OH-dG contents in plasma. The anti-oxidation enzyme activity in erythrocytes was also increased, enzymes concerned including superoxide dismutase, glucose-6-phosphate dehydrogenase, catalase and glutathione peroxidase. In addition, the intake of Ganoderma lucidum was proved effective to bring subjects with high GPT and GOT back to normal range. The abdominal ultrasound examination also showed improvement on fatty liver.

碩士
國立陽明大學
生物醫學資訊研究所
98
Ling-Chi (Ganoderma lucidum), a chinese herb, The recent studies have revealed that polysaccharides, triterpenes, and some microelements in Ling-Chi, that has been reported to be effective in the treatment of neoplasia, hypertension, and hepatopathy. A whole genome sequence project for Ganoderma lucidum is well-studied to understand of biosynthesis of the secondary metabolites and some microelements. Next-generation high-throughput DNA sequencing techniques are opening whole genome sequence assembled. Ling-Chi whole genome project sequenced using a combination of conventional capillary sequencer and new-generation sequencing technology, and was conducted by a collaboration of the team of the Veteran General Hospital-Taipei（VGH-Tapei）and the National Yang-Ming University（NYMU）from 2001 to present. The genome of Ling-Chi is about 45Mb in size and it contains 13 chromosomes. We predicted 14,052 genes by using gene prediction tool Genemark-ES in Ling-Chi. Moreover, these genes we used tentative unique genes (TUG) sequences applied to verify the gene structure predicted by prediction programs. The possible functions and reaction pathways of these predicted genes were annotated by using the sequence similarity to known proteins. And then, we identified some genes that might be involved in the terpenoid backbone and steroid biosynthesis by KEGG in Ling-Chi. In additional to discussing the genome assembly correctness of Ling-Chi genome, using by mate-pair short reads and TUG sequences can be applied to provide the order of contigs in genomic scaffold.

碩士
中國醫藥大學
中國藥學暨中藥資源學系碩士班
102
Ganoderma lucidum, a popular traditional Chinese medicinal herb, has been used to treat or prevent disease for thousand years in Asia. Ganoderic acids (GAs) are one of major biologically active components and show pharmacological activities such as anti-oxidant, anti-tumor, anti-virus and anti-inflammation. However, the regulation of GAs biosynthesis is poorly understood. This study investigates the role of apoptosis signaling on GAs biosynthesis. The drugs which induced apoptosis in mammalian cells or fungi cells, such as acetic acid (AA), menadione (Vit K3), sodium chloride (NaCl), valproic acid (VPA), zinc chloride (ZnCl2), were used to treat the mycelium cell of G. lucidum. Biomass and GAs production, and cell apoptosis were evaluated. The content of total GAs was analyzed by high performance liquid chromatography (HPLC). The cell apoptosis was identified by terminal deoxynucleotidyl transferase mediated dUTP nick end labeling assay (TUNEL) and nuclear morphology change. All treated drugs reduced biomass and induced GAs production. As compared with control, maximum total GAs accumulation were 1.79-fold, 2.39-fold, 1.67-fold, 2.06-fold and 1.81-fold induced by 40 mM AA, 2 mM Vit K3, 1 M NaCl, 2.5 mM VPA and 10.8 mM ZnCl2, respectively, for 4 days incubation. In addition, Vit K3 (0.5 mM to 2 mM), VPA (0.8 mM to 1.2 mM) and ZnCl2 (0.8 mM to 1 mM) induced apoptosis which were identify by positive TUNEL stain and condensed nuclear morphology. This data indicates that Vit K3, VPA and ZnCl2 induced GAs biosynthesis during cell apoptosis.

碩士
靜宜大學
應用化學研究所
98
Liposomes are spheres made of lipid bilayers, enclosing aqueous volumes. The advantage of liposomal delivery is that separates the functions required for drug delivery. During the 1990s, a number of lipsome-based drugs reached the market in the U.S. and Europe and there has been an explosion of new procedures and techniques in the field. Ganoderma lucidum is used as an edible medicinal material in Chinese medicine. In recent years, many scientific researches have shown G. lucidum has a wide variety of functions. The research focuses on combining liposome with G. lucidum by using thin-film method in order to prepare the liposomes encapsulated G. lucidum extract, and to evaluate its physical stability concerning the particle size and encapsulation efficiency. Based on this study, we found that the optimal drug/lipid ratio still remains high encapsulation efficiency above 61%, and particle size is less than 15.24 μm after three months storage at 4℃.

碩士
嘉南藥理科技大學
生物科技系暨研究所
91
The purpose of this study is to isolate and purify the tyrosinase inhibitor from Ganoderma lucidum. Tyrosinase is a key enzyme in melanogenesis, therefore tyrosinase inhibitor can regulate the enzyme activity of tyrosinase and prevent the undesirable effect of skin as well as browning reaction of food. The tyrosinase inhibitor from Ganoderma lucidum was extracted by various extraction media, and that extracted by 25mM sodium phosphate buffer pH6.8 showed the highest extraction efficiency, followed by methanol?z n-butanol?z n- hexane and ethyl acetate . The inhibitor was then stored at -18℃ and 4℃, the relative inhibition activity of tyrosinase were 98% and 88% after 6 month storage, respectively. The tyrosinase inhibitor from Ganoderma lucidum was further purified with　Amberlite XAD-7?z Sephadex G-25 and Superdex Peptide column chromatography, and the recovery was 83%?z 50% and 15%, respectively; the inhibitory concentration caused 50% inhibition (IC50) was 5.8?z 3.19 and 0. 15mg/ml, respectively. The inhibition activity of tyrosinase was 47% when preincubated with the inhibitor for 0 minute while 100% for 18 hour, indicating a slow binding behavior. The optimal temperature of the inhibition activity of tyrosinase treated with the inhibitor was 45℃-55℃; optimal pH 5. 0. The inhibitor was stable at pH 4.0-9.0. In Kinetic study, the inhibitor showed mixed types and the inhibitory constant (Ki) 1.1mg/ml using L-DOPA as substrate, while mixed types and Ki 0.3mg/ml using L-tyrosine as substrate. Moreover, molecular mass of the partially purified tyrosinase inhibitor from Ganoderma lucidum was 0.3kDa examined by Superdex Peptide column gel filtration. The tyrosinase inhibitor from Ganoderma lucidum may contain sugar residue or amino group, according to the positive reaction with Fehling reagent and ninhydrin.

by Ma Suet-yee.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2002.
Includes bibliographical references (leaves 119-131).
Abstracts in English and Chinese.
Acknowledgment --- p.i
Abstract --- p.ii
摘要 --- p.iv
Table of Contents --- p.vi
List of Tables --- p.x
List of Figures --- p.xii
Chapter Chapter 1 --- Introduction --- p.1
Chapter 1.1 --- Ganoderma lucidum --- p.1
Chapter 1.1.1 --- History of Ganoderma lucidum --- p.1
Chapter 1.1.2 --- Classification --- p.1
Chapter 1.1.3 --- Macroscopic and microscopic structure --- p.2
Chapter 1.1.4 --- Ganoderma lucidum as a pathogen --- p.3
Chapter 1.1.5 --- Availability of tree hosts in Hong Kong --- p.4
Chapter 1.1.6 --- Medicinal effects --- p.5
Chapter 1.2 --- Study of Populations in Fungi --- p.6
Chapter 1.2.1 --- Definition of Population --- p.6
Chapter 1.2.2 --- Study of Fungal Populations --- p.7
Chapter 1.2.3 --- Techniques for Population Studies in Fungi --- p.7
Chapter 1.2.3.1 --- Somatic Incompatibility Test
Chapter 1.2.3.2 --- Isozyme Analysis
Chapter 1.2.3.3 --- Restriction Fragment Length Polymorphisms (RFLPs)
Chapter 1.2.3.4 --- Polymerase Chain Reaction (PCR) Amplification
Chapter 1.3 --- Mitochondrial DNA (mt-DNA) in Fungi --- p.14
Chapter 1.3.1 --- Inheritance in mt-DNA --- p.15
Chapter 1.3.2 --- Mitochondrial DNA in Population Studies --- p.15
Chapter 1.3.2.1 --- Mitochondrial small-subunit (mt-SSU) rDNA
Chapter 1.3.2.2 --- Cytochrome oxidase 3 (cox3)
Chapter 1.4 --- Biodiversity study on Ganoderma species --- p.19
Chapter 1.5 --- Environment Pollutants in Hong Kong --- p.20
Chapter 1.5.1 --- Air quality in Hong Kong --- p.20
Chapter 1.5.2 --- Soil quality in Hong Kong --- p.20
Chapter 1.5.3 --- Toxicity of pollutants --- p.23
Chapter 1.5.4 --- Accumulation of heavy metals by G. lucidum --- p.26
Chapter 1.6 --- Objectives of Study --- p.27
Chapter 1.7 --- Project Strategies --- p.28
Chapter 1.7.1 --- Survey on distribution and collection of Ganoderma lucidum in Hong Kong --- p.28
Chapter 1.7.2 --- Genetic divergences of G. lucidum mitochondrial genes --- p.28
Chapter 1.7.3 --- Contaminations on field collected G. lucidum --- p.29
Chapter 1.8 --- Significance of Study --- p.29
Chapter Chapter 2 --- Materials and Methods --- p.30
Chapter 2.1 --- Collection of Ganoderma lucidum in Hong Kong --- p.30
Chapter 2.2 --- Tissue Isolation --- p.30
Chapter 2.3 --- Somatic Incompatibility Test --- p.36
Chapter 2.4 --- Molecular Identification --- p.40
Chapter 2.4.1 --- Extraction of DNA --- p.40
Chapter 2.4.2 --- Gel electrophoresis --- p.41
Chapter 2.4.3 --- Strain authentication by arbitrarily primed polymerase chain reaction (APPCR) --- p.41
Chapter 2.4.4 --- PCR of mt-SSU rDNA and --- p.43
Chapter 2.4.5 --- Sequencing of mt-SSU rDNA and cox3 --- p.44
Chapter 2.4.6 --- Comparison of G. lucidum complex with other Ganoderma and related species
Chapter 2. 4.7 --- Phylogenetic analyses --- p.46
Chapter 2.5 --- Investigation of pollutants in Ganoderma lucidum collected in Hong Kong --- p.46
Chapter 2.5.1 --- Metal analysis --- p.48
Chapter 2.5.1.1 --- Acid digestion
Chapter 2.5.1.2 --- Statistical analysis
Chapter 2.5.2 --- Organic pollutant analysis --- p.49
Chapter Chapter 3 --- Result --- p.52
Chapter 3.1 --- Collection of Ganoderma lucidum in Hong Kong --- p.52
Chapter 3.1.1 --- Field observation --- p.52
Chapter 3.1.2 --- Macroscopic characteristics --- p.52
Chapter 3.1.3 --- Microscopic characteristics --- p.53
Chapter 3.2 --- Somatic Incompatibility Test --- p.56
Chapter 3.3 --- DNA fingerprints by Arbitrarily-Primed PCR --- p.57
Chapter 3.4 --- Sequencing of mt-SSU rDNA region of G. lucidum and related species --- p.60
Chapter 3.4.1 --- Genetic variability in mt-SSU rDNA region of G. lucidum --- p.60
Chapter 3.4.2 --- mt-SSU rDNA region of G. lucidum and other related species --- p.61
Chapter 3.4.3 --- Phylogenetic analysis of mt-SSU rDNA region --- p.61
Chapter 3.5 --- Sequencing of cox3 region --- p.71
Chapter 3.5.1 --- Genetic variability in cox3 region of G. lucidum --- p.71
Chapter 3.5.2 --- cox3 region of G. lucidum and other related species --- p.72
Chapter 3.5.3 --- Phylogenetic analysis of cox3 region --- p.72
Chapter 3.6 --- Metal content of field G. lucidum --- p.82
Chapter 3.7 --- Organic pollutants in field collected G. lucidum --- p.90
Chapter Chapter 4 --- Discussion --- p.93
Chapter 4.1 --- Collection of Ganoderma lucidum in Hong Kong --- p.93
Chapter 4.1.1 --- Differentiation of G. lucidum and related species in the G lucidum species complex --- p.93
Chapter 4.1.2 --- Field observation --- p.94
Chapter 4.2 --- Biodiversity of populations of G. lucidum in Hong Kong --- p.95
Chapter 4.2.1 --- Individualism of G. lucidum --- p.95
Chapter 4.2.2 --- Genetic variability in mt-SSU rDNA region of G. lucidum --- p.96
Chapter 4.2.3 --- Genetic variability in cox3 region of G. lucidum --- p.98
Chapter 4.2.4 --- Lineages of G. lucidum collected in Hong Kong --- p.100
Chapter 4.2.5 --- Cryptic phylogenetic species --- p.101
Chapter 4.3 --- Contamination of field collected Ganoderma lucidum in Hong Kong --- p.106
Chapter 4.3.1 --- Metal contents in field collected G. lucidum in Hong Kong --- p.106
Chapter 4.3.1.1 --- Metal contents of G. lucidum fruiting bodies collected at each site
Chapter 4.3.1.2 --- General discussion of metals
Chapter 4.3.1.3 --- Consumption of field collected G. lucidum fruiting bodies
Chapter 4.3.2 --- Comparison of metal contents between field collected Hong Kong G. lucidum with other mushrooms collected from other places --- p.112
Chapter 4.3.3 --- Survey of organic pollutants in field collected G. lucidum in Hong Kong --- p.113
Chapter Chapter 5 --- Conclusion --- p.116
Chapter Chapter 6 --- Further investigation --- p.118
Chapter Chapter 7 --- Reference --- p.119

by Cheung Ka Wan.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2001.
Includes bibliographical references (leaves 121-128).
Abstracts in English and Chinese.
Chapter Chapter 1 - --- Introduction --- p.1
Chapter 1.1 --- Plant Pathogens --- p.1
Chapter 1.1.1 --- Virus --- p.2
Chapter 1.1.2 --- Viroids --- p.2
Chapter 1.1.3 --- Bacteria --- p.3
Chapter 1.1.4 --- Fungi --- p.3
Chapter 1.1.5 --- Mycoplasma like organisms --- p.3
Chapter 1.1.6 --- Nematodes --- p.4
Chapter 1.1.7 --- Insects --- p.4
Chapter 1.1.8 --- Mammals --- p.4
Chapter 1.2 --- Pathogenicity --- p.5
Chapter 1.3 --- Disease Development --- p.6
Chapter 1.3.1 --- Primary infection --- p.6
Chapter 1.3.2 --- Penetration to host --- p.6
Chapter 1.3.2.1 --- Entry through wounds ^
Chapter 1.3.2.2 --- Entry through natural openings --- p.8
Chapter 1.3.2.3 --- Direct penetration
Chapter 1.3.3 --- Colonization of pathogen --- p.9
Chapter 1.3.4 --- Mechanisms of attack --- p.9
Chapter 1.3.5 --- Symptom expression --- p.11
Chapter 1.3.6 --- Spread of disease --- p.11
Chapter 1.4 --- Detection of Pathogen --- p.12
Chapter 1.4.1 --- Traditional diagnostic methods --- p.12
Chapter 1.4.2 --- Molecular diagnostic methods --- p.13
Chapter 1.4.3 --- Advantages of molecular diagnostic tools over traditional detection methods --- p.14
Chapter 1.4.4 --- Sensitivity of molecular diagnostic tools --- p.14
Chapter 1.4.5 --- Polymerase chain reaction (PCR) --- p.15
Chapter 1.4.5.1 --- Mechanism of PCR --- p.16
Chapter 1.4.5.2 --- Application of PCR --- p.17
Chapter 1.4.6 --- Designation of specific primers in pathogen detection --- p.17
Chapter 1.4.6.1 --- Nuclear ribosomal DNA genes --- p.18
Chapter 1.4.6.2 --- Sequencing of ITS regions of rDNA --- p.19
Chapter 1.5 --- Ganoderma lucidum Complex --- p.19
Chapter 1.5.1 --- History of Ganoderma lucidum complex --- p.19
Chapter 1.5.2 --- Classification --- p.20
Chapter 1.5.3 --- Macroscopic and microscopic structure --- p.21
Chapter 1.5.4 --- Species identification in G. lucidum complex --- p.22
Chapter 1.5.5 --- Ganoderma species in Hong Kong --- p.23
Chapter 1.5.6 --- Act as pathogen --- p.25
Chapter 1.5.7 --- Availability of tree hosts in Hong Kong --- p.25
Chapter 1.5.7.1 --- Acacia confusa --- p.26
Chapter 1.5.7.2 --- Listea cubeba --- p.26
Chapter 1.5.7.3 --- Leucaena leucocephala --- p.27
Chapter 1.5.8 --- Disease control for Ganoderma lucidum --- p.27
Chapter 1.6 --- Aims of Study --- p.29
Chapter 1.7 --- Significance of the Study --- p.29
Chapter 1.8 --- Project Strategies --- p.30
Chapter 1.8.1 --- Survey on Ganoderma lucidum complex in Hong Kong --- p.30
Chapter 1.8.2 --- Artificial infection --- p.30
Chapter 1.8.3 --- Detection of pathogen --- p.30
Chapter Chapter 2 - --- Materials and Methods --- p.31
Chapter 2.1 --- Collection of Ganoderma lucidum Species Complex in Hong Kong --- p.31
Chapter 2.2 --- Tissue Isolation --- p.31
Chapter 2.3 --- Molecular Identification --- p.45
Chapter 2.3.1 --- Extraction of DNA --- p.45
Chapter 2.3.2 --- Gel Electrophoresis --- p.45
Chapter 2.3.3 --- Sequencing of ITS 1 and ITS2 --- p.46
Chapter 2.3.4 --- Comparison of G. lucidum complex with other Ganoderma and related species --- p.48
Chapter 2.3.5 --- Strain authentication by arbitrarily primed polymerase chain reaction (APPCR) --- p.49
Chapter 2.4. --- Mating Compatibility for Species Delimitation --- p.49
Chapter 2.4.1 --- Protoplast isolation --- p.49
Chapter 2.4.2 --- Mon-Mon mating --- p.50
Chapter 2.4.3 --- Di-Mon mating --- p.50
Chapter 2.5 --- Preparation of Samples for Scanning Electron Microscope (SEM) --- p.51
Chapter 2.6 --- Cytological Studies of Basidiocarps of G. lucidum --- p.52
Chapter 2.7 --- Pathogenicity Study --- p.53
Chapter 2.7.1 --- Growth and spread of G. lucidum in soil --- p.53
Chapter 2.7.2 --- Colonization of G. lucidum on different organs of plants --- p.53
Chapter 2.7.2.1 --- Determination of dry weight loss --- p.53
Chapter 2.7.2.2 --- Chitin assay --- p.54
Chapter 2.7.3 --- Artificial infection to tree seedlings --- p.54
Chapter 2.7.3.1 --- Artificial infection of vegetative mycelia --- p.54
Chapter 2.7.3.2 --- Artificial infection with basidiospores --- p.56
Chapter Chapter 3 - --- Results --- p.57
Chapter 3.1 --- Collection of Ganoderma lucidum Complex in Hong Kong --- p.57
Chapter 3.1.1 --- Macroscopic characteristics --- p.57
Chapter 3.1.2 --- Microscopic characteristics --- p.57
Chapter 3.1.3 --- G. lucidum under scanning electron microscopy --- p.59
Chapter 3.2 --- Field Observation --- p.62
Chapter 3.3 --- Sequencing of ITS Region of G. lucidum Complex and Related Species --- p.64
Chapter 3.3.1 --- ITS 1 Region of G. lucidum --- p.66
Chapter 3.3.2 --- ITS 2 Region of G. lucidum --- p.68
Chapter 3.3.3 --- Relationship between Ganoderma and related species --- p.71
Chapter 3.4 --- Species Delimitation of G. lucidum --- p.74
Chapter 3.4.1 --- Arbitrarily-Primed PCR --- p.74
Chapter 3.4.2 --- Di-Mon mating --- p.77
Chapter 3.5 --- Pathogenicity of G. lucidum --- p.81
Chapter 3.5.1 --- Growth and spread in soil --- p.81
Chapter 3.5.2. --- Preference in colonization on different organs of plants --- p.81
Chapter 3.5.3 --- Artificial infection --- p.87
Chapter Chapter 4 - --- Discussion --- p.98
Chapter 4.1 --- Ganoderma lucidum Complex in Hong Kong --- p.98
Chapter 4.1.1 --- Macroscopic and microscopic characteristics of G. lucidum complex and related species --- p.98
Chapter 4.1.2 --- Cytological studies --- p.99
Chapter 4.1.3 --- Field observation --- p.100
Chapter 4.1.4 --- Sequences of ITS regions of G. lucidum complex and related species --- p.101
Chapter 4.1.5 --- Species identification within G. lucidum --- p.104
Chapter 4.2 --- Pathogenicity Test for G. lucidum --- p.108
Chapter 4.2.1 --- Growth and spread of G. lucidum --- p.108
Chapter 4.2.2 --- Colonization of G. lucidum on plants --- p.110
Chapter 4.2.3 --- Artificial infection by G. lucidum --- p.111
Chapter 4.3 --- Further Investigation --- p.116
Chapter Chapter 5 - --- Summary --- p.118
Chapter Chapter 6- --- Conclusion --- p.120
References --- p.121

碩士
國立臺灣大學
食品科技研究所
94
Ganoderma lucidum is one of the most well known Chinese medicine in ancient times and is considered as the most valuable plant for health and longevity promotion. In this study, we investigated the effects of Ganoderma lucidum liquid fermentation broth on immunomodulatory activities in vitro and in vivo. After an in vitro experiment, we selected the most active fermentation product for experiments on non-specific immune response in BALB/c mice. In addition, we observed the MNC proliferation effect and performed the proteomic analysis of the protein expression by human mononuclear cells (MNC) treated with ethanol precipitated Ganoderma lucidum polysaccharide. Results show that Ganoderma lucidum fermentation product GL-93-12-21 stimulates human mononuclear cells to secrete cytokines and inhibit the human myelocytic lymphoma U937 cell growth. Moreover, the ethanol precipitated polysaccharide stimulats NO and TNF-α production in RAW 264.7 macrophage, and the polysaccharide characteristics are similar with bioactive polysaccharides that have been reported. In vivo studies showed that oral administration of Ganoderma lucidum fermentation product GL-93-12-21 on BALB/c mice significantly (P＜0.05) increases the secretion of IgG, the proliferation of T- and B-lymphocyte, the level of TNF-α、IL-2 and IFN-γ production, the phagocytosis effect of monocytes and the cytotoxicity of nature killer cells. Results also show that Ganoderma lucidum polysaccharide stimulates the proliferation of human mononuclear cells and changes the protein expression. There are 19 protein spots (1 down-regulated and 18 up-regulated) differentially expressed more than 1.5-fold, which was identified by quantitative image analysis and matrix assisted laser desorption ionization-quadrupole-time of flight (MALDI-Q-TOF) mass spectrometry. The Swiss-Prot database search indicated that increased expression of cytoskeleton and ribonucleoprotein is associated with the immune cell activation and proliferation. Furthermore, the increase of annexin A6 and chaperon protein expression, may play a part in activating of B lymphocyte and assembling of B cell receptor.

碩士
國立中興大學
植物學系
87
Ganoderma lucidum is an important medical fungus. It seems be a trend that was studied at the molecular level recently. Transformation system helps studying its molecular genetics and may apply to strain improvement. So we attempt to build up the gene target transformation system based on Hygromycin B resistant gene hph of pAN7-1, a broad range host vector of fungi, as a selection marker. In this study, we cloned 3 homologous DNA fragments of G. lucidum by PCR. Target vectors were constructed by inserting these fragments at HindIII site of pAN7-1, and named pGL400、pGL600、pGL1k individually. PEG/CaCl2 transformation was mediated with 10 g plasmid DNA and protoplasts preparing from monokaryotic mycelia. The transformation frequency was 16.4、11.5、13.5 transforants per g DNA respectively for pGL400、pGL600、pGL1k and higher than pAN7-1,which was 3.5 transforants per g DNA. And most transforants harboring HygBr phenotype still after several subcultures without any selection pressure. The evidence of PCR product and HygBr phenotype proved that the plasmids could be transformed into G. lucidum successfully. Penotypic analysis of these transforants shows the differences in the ability to resist Hygromycin B, colony morphology and growth rate from each. And the results of pGL400 transforants with high proportion of morphological aberration implicated that might involved in the gene function of 400 bp. In addition, we analysis the cellobiohydrolase activity of the pGL1k transforants. And we found that a tranforant TnpGL1kt3 harboring very low CBH activity maybe a cbhI mutant.

碩士
國立臺灣大學
醫事技術學研究所
89
Ganoderma spp. has a long history of use in folk medicine. Many bioactive compounds have been identified from their fruiting body, spore and mycelium. In this thesis, we want to study the anticancer effects and the mechanisms of the extracts of Ganoderma spp. The hot water extract of Ganoderma lucidum mycelium was fractionated into triterpene fraction and polysaccharide fraction. The triterpene fraction was studied in vitro aimed at the differential growth effects on human normal liver cells and hepatoma cells. The polysaccharide fraction was investigated in animal model aimed at inhibiting effects on DMBA-induced carcinogenesis of oral epithelium. Results showed that the triterpene fraction exhibited stronger growth inhibition on hepatoma cells than on normal liver cells. Triterpene fraction caused a decrease in protein levels of cell growth regulative proteins, PKC-a and its downstream effectors, c-myc and NF-kB. This suppression resulted in growth inhibition of hepatoma cells. Cell cycle analysis by flow cytometry showed that the hepatoma cells were arrested at G2-M phase. These changes on hepatoma cells, but not on normal liver cells lead to differential growth inhibition. In hepatoma cells the triterpene fraction also induced cell apoptosis by increasing protein levels of pro-apoptotic protein, p53, MAPK and caspase and decreasing anti-apoptotic protein, Bcl-2. On the other hand, exposure to triterpene fraction caused decreases in the activity of superoxide dismutase (SOD) and the level of cellular protein-SH, and an increase in H2O2 level. Reduction of antioxidant components by triterpene fraction may also be a reason that cause cell death. In syrian hamster, intraperitoneal injection of polysaccharide fraction can inhibit neoplasm of oral epithelium induced by DMBA. The efficacy was shown by less or smaller tumor formation, and less malignant characteristies judged from fluorescence spectra and histopathology of tissues. We further discovered that the polysaccharide fraction would decrease the activity of pro-metastasis protein, matrix metalloproteinase-9 (MMP-9). The data showed that reduction of MMP-9 might retard the invasive activity of cancer. In conclusion, the triterpene fraction was effective in inhibiting hepatoma cell growth by reducing cell cycle progression and the growth regulative protein, while has little effects on normal liver cell. Furthermore, this fraction would lead to cell death by inducing cell apoptosis and increasing cellular oxidative stress in hepatoma cells. The polysaccharide fraction exhibited stronger anti-carcinogenesis effect in vivo. This study would be worth to probe into the feasible way for drug treatments in cancer.

碩士
國立陽明大學
生物資訊研究所
94
Ling-Chi (Ganoderma lucidum) is one of the traditional Chinese herbs. The recent studies have revealed that polysaccharides, triterpenes, and some microelements in Ling-Chi have several interesting effects, such as anti-tumor, hypoglycemic, hypocholesterol, and immuno-stimulation. The genome and expressed sequence tag (EST) analyses are useful in identifying the genes involved in the biosynthesis of polysaccharides or triterpenes. Ling-Chi genome project have sequenced the draft sequence by shotgun method. In order to discover the expressed genes, the EST project has completed the sequencing of 47,122 EST sequences in 2005. The redundancy of the EST sequences was removed by two steps. The EST sequences were assembled and then be compared with the known protein sequences of other species. Those sequences that match the same protein were merged and got 9,697 tentative unique genes (TUG-1). The possible functions and reaction pathways of these TUG’s were annotated by using the sequence similarity to known proteins. On the basis of literature survey and sequence analysis, I identified 46 genes that might be involved in the polysaccharide biosynthesis in Ling-Chi. Moreover, 20 genes among the 46 genes, which are very similar to the known genes in the database, were annotated further. The gene structure, the predicted protein sequences, the putative protein domains of these genes were summarized for functional studies. In additional to annotating the function of Ling-Chi genes, EST sequences can also be applied to verify the gene structure predicted by programs, to estimate the gene numbers, to provide the order of genomic contigs, and to find the putative paralogous genes of Ling-Chi. Selected examples are discussed to demonstrate the feasibility of such approaches.

碩士
國立臺灣大學
食品科技研究所
106
Ganoderma formosanum and Ganoderma lucidium, Lingzhi, are traditional Chinese medicine and are considered as healthy food. The bioactive components in Lingzhi are (1, 3；1, 6)-β-D-glucans and ganoderic acids. This study aims to investigate that the effect of the static culture, the submerged culture and the two-stage fermentation on the production of the (1, 3；1, 6)-β-D-glucans and ganoderic acids of Ganoderma formosanum and Ganoderma lucidum. The biomass production, polysaccharide in mycelium and broth, the content and the degree of the branch of (1,3；1,6)-β-D-glucans and the structure of ganoderic acids of Ganoderma formosanum and Ganoderma lucidum were compared. In this study, the (1,3)-β-D-glucanase was used to investigate the content and the degree of the branch of (1,3；1,6)-β-D-glucans. Liquid chromatography-tandem mass spectrometry was used to analyze ganoderic acids. The structure properties of ganoderic acids were identified by accurate m/z (-2.33-2.33 ppm), MS/MS spectra and UV spectra. Standard addition method was used to analyze the concentration of ganoderic acids. Relative quantification method was used to analyze the ganoderic acids without an available commercial standard. In this research, the result shows that static culture produces more ganoderic acid than submerge culture. However, shaking culture produces more biomass and polysaccharide. Two-stage fermentation can enhance the yield of the target compound. The mycelium of Ganoderma formosanum has maximum biomass by static culture. At 10 days, Ganoderma formosanum has the maximum yield of polysaccharides in the mycelium and the broth. By shaking culture, the maximum biomass of the mycelium of Ganoderma lucidum is at 15 days and the maximum yield of the polysaccharide of Ganoderma lucidum is at 10 days but there is no significant difference in the broth. At 30 days, by static culture, the content of (1,3；1,6)-β-D-glucan of Ganoderma formosanum mycelia is more than Ganoderma lucidum. The degree of the branch of (1,3；1,6)-β-D-glucan of Ganoderma formosanum mycelia is between 0.2-0.33. Ganoderma lucidum only produces (1,3；1,6)-β-D-glucan in static culture and the degree of the branch of (1,3；1,6)-β-D-glucan is lower than 0.2-0.33. Ganoderma lucidum contains more mycelia and polysaccharides than Ganoderma formosanum. However, the percentage of (1,3；1,6)-β-D-glucan in the polysaccharides of Ganoderma formosanum is more than Ganoderma lucidum. Ganoderma lucidum and Ganoderma tsugae fruiting body were used as QC sample. 27 kinds of ganoderic acids were found in QC sample. In this study, the mycelium of Ganoderma formosanum in static culture 30 days has more ganoderic acid H than Ganoderma lucidum. The static culture and shaking culture have no significant effect on the production of Ganoderic acid H of Ganoderma lucidum.

碩士
國立中央大學
化學工程與材料工程學系
103
Ganoderma lucidum is one of the medicinal mushrooms. The most attractive characteristic of the medicinal mushrooms is its anti-tumor effect. Ganoderma lucidum has gained wide popularity as a health food and became the most valuable mushroom in Taiwan. However, because of its host specificity, slow growth rate and rarity in nature, the fruiting bodies of Ganoderma lucidum have become expensive mushrooms in recent years. Thus, investigators have exerted their efforts to prepare this mushroom from submerged culture. The main objectives of this proposal are to develop two-stage fermentation system. In first stage, we make the Ganoderma lucidum grow in high speed for 8 days, and Saccharomyces cerevisiae was added to the fermentation broth in second stage with aerated or static process to enhance ganoderic acids production. The amount of ganoderic acids reached 171.5 mg/g DW by using static operation with 5%(v/v) of inoculation level of the Saccharomyces cerevisiae in 250 ml shake flask. A novel two stage operation to optimize triterpenoids production of a co-culture was proposed and successfully demonstrated, reaching highest ganoderic acids content obtained in this study.

碩士
臺北醫學大學
生物醫學材料研究所
96
Cancer is the leading cause of deaths worldwide. Lately, Chinese herbal medicine has been used increasingly in allopathic medicine. Ganoderma lucidum, also known as “LingZhi”, is a woody Basidiomyceyes mushroom which belongs to the Ganodermaceae family. Previous studies demonstrated that Ganoderma lucidum ethanol extracts exerted immunomodulating, antioxidative, anticancer activity and cytotoxicity to cancer cells. The objective of this research was to identify the bioactive compounds in ethanol extracts of Ganoderma lucidum which contain anti-tumor activities. In our study, we demonstrated that Ganoderma lucidum ethanol extracts inhibited proliferation of human cervical cancer, HeLa cells, in a dose-dependent fashion. Ganoderma lucidum ethanol extracts treatment led to an increase number of HeLa cells in sub-G1 phase as well as the induction of apoptosis. Ganoderma lucidum ethanol extract also increased reversion frequency by decreasing the expression of oncoprotein E7 and alkaline phosphatase activity in HeLa cell. Human non-small cell lung cancer cell line, H441GL cells, which contains a firefly luciferase gene was used for non-invasive in vivo tumor volume monitoring and assessement of novel therapeutics. In this part, we demonstrated that Ganoderma lucidum ethanol extract induced the expression of cyclin E protein and caused H441GL cells to arrest in G0/G1 phase thereby inhibiting their proliferation. In vivo animal experiments, we demonstrated that feeding Ganoderma lucidum treatment yielded a much less aggressive tumor growth than the control group. Although further studies are requied, the present work suggests that Ganoderma lucidum may have beneficial effects in treating both cervical cancer cells, HeLa, and NSCLC such as H441GL carcinoma. Non-invasion bioluminescent imaging system provides a functional way of monitoring tumor volume in animal model. Ganoderma lucidum may raise the possibility of continuing management of some cancers as a chronic condition in which the malignancy is constrained.

碩士
東海大學
化學工程學系
85
In this research ,the mycelia of Ganoderma lucidum were grown immobilizedlyon a polyurethane foam for the production of polysacchraide.The main experiments were carried out in 250ml shake flasks,packed columns ,a stirredreactor and a air-lift reactor.In suspended shake-flask cultures,the maximum concentration of mycelia & polysaccharide were about 800mg/100ml & 60mg/100ml.In contrast,the value were 1300mg/100ml & 50mg/100 ml in immobilized cultures. The results indicated that the mycelia could grow better on the surface ofpolyurethane foam materials. In addition, the viscosity of the broth was lower, due to the fact that produced polysaccharide may adhere to the surface of foam material and formed a gel layer. The percentage of polysaccharide present in gel form could increase with time , and was more than 50% on the14th day. In a repeated-batch test using a packed column, the immobilized mycelia showed the capability of producing polysaccharide continuously.When th immobilized culture was carried out in a stirred tank bioreactor , the foam could be decomposed under the condition of high shear stress. Theconcentration of polysaccharide in the culture using a air-lift bioreactorcould reach to 60mg/100ml on 9th day, much rapidly than the culture in shakeflasks.

碩士
靜宜大學
食品營養研究所
94
Polysaccharides were extracted from the fruit body and mycelium of Ganoderma lucidum with 100 ℃ hot water and dried by hot air、vacuum and freeze drying, respectively. Rheological properties of the resulted in Ganoderma lucidum polysaccharides were studied using Canon-Fensky glass capillary viscometer and dynamic rheometer. It was found that the Huggins’ constant of Ganoderma lucidum polysaccharides was 0.39-0.48 and 0.59-0.93 when DMSO and H2O were used as solvents respectively. It seems that DMSO is a better solvent than H2O for Ganoderma lucidum polysaccharides. Plysaccharides extracted from the fruit body and mycelium of Ganoderma lucidum have average molecular weights of 1.26×106 and 1.38×106 respectively, and a Mark – Houwink constant in the range of 1.32-1.66. It was found the intrinsic viscosity of Ganoderma lucidum polysaccharides decreased with increasing ionic strength. The salt tolerance was 5.30-5.45 ml M0.5 /g and chain stiffness parameter was about 0.062-0.073. The specific viscosity versus concentration plot exhibited a power-law dependence on concentration. The coil overlap parameter of fruit body polysaccharides was 0.31 to 0.35,and the exponent b ranged from 0.98 to 1.06 in the dilute domain and from 1.12 to 1.25 in the semi-dilute domain. The coil overlap value of the polysaccharides extracted from mycelium was about 1.38 - 2.09 and the slope of the diluted and entangled domain were 1.10 - 1.43 and 1.72 - 2.49 respectively. Drying method did not show a significant effect on the rheological properties. During steady shear measurements, Ganoderma lucidum polysaccharides showed shear thinning behavior, which could be analyzed by a power law model. The consistency index of Ganoderma lucidum polysaccharides increased, and the flow behavior index decreased with increasing concentration. In general, the polysaccharides extracted from mycelium showed higher viscosity than that from fruit body. During dynamic shear measurements, within the frequency range tested, the storage modulus were greater than loss modulus, and increased with increasing concentration.

Dissertação de Mestrado Integrado em Engenharia Química apresentada à Faculdade de Ciências e Tecnologia
Os cogumelos têm vindo a ser incluídos de forma cada vez mais frequente na alimentação humana, assumindo um papel importante, quer devido às suas características nutricionais e organoléticas, quer devido às diferentes formas de consumo, despertando sensações únicas, devido ao seu aroma e sabor que colocam e despertam em cada prato. Por outro lado, são também muito consumidos, devido às suas propriedades medicinais extraordinárias, sendo cada vez mais alvo de muitos estudos realizados neste âmbito. Estudos científicos e médicos demonstram as propriedades medicinais dos compostos extraídos de cogumelos para a prevenção das doenças mais diversas, sendo cada vez mais frequentes na prevenção e tratamento do cancro (Zaidman et al., 2005; Lemieszek et al., 2013). O objetivo deste trabalho consiste na caracterização química de quatro espécies de cogumelos, Boletus edulis, Tricholoma equestre, Ganoderma lucidum e Ganoderma lingzhi e no estudo do efeito de um dos seus compostos na atividade neuronal. Neste estudo, investigou-se a ação de polissacarídeos extraídos dos cogumelos na atividade antioxidante de uma zona sináptica. Os experimentos consistem na adição de diferentes concentrações de tais extratos na região CA3 do hipocampo do cérebro no sistema sináptico de fibras musgosas usando fatias de cérebro de 400 μm de espessura.----------------------------------------------------------------------------------------------------------------------------------------------------------------
Mushrooms have been increasingly included in human food, taking on an important role, either due to their nutritional and organoleptic characteristics or due to the different forms of consumption, arousing unique sensations, due to their aroma and flavor put and wake up on each plate. On the other hand, they are also very consumed, due to their extraordinary medicinal properties, being more and more the target of many studies carried out in this scope. Scientific and medical studies demonstrate the medicinal properties of compounds extracted from mushrooms for the prevention of the most diverse diseases, being more and more frequent in the prevention and treatment of cancer (Zaidman et al., 2005; Lemieszek et al., 2013). The aim of this work is the chemical characterization of four mushroom species, Boletus edulis, Tricholoma equestre, Ganoderma lucidum and Ganoderma lingzhi, and to study the effect of one of its compounds on neuronal activity. In this study, we investigated the action of polysaccharides extracted from mushrooms on the antioxidant activity of a synaptic zone. The experiments consist of adding different concentrations of such extracts to the CA3 region of the brain hippocampus in the moss-synaptic system using 400 μm thick brain slices.-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------

Li Pik Ha Ivy.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2006.
Includes bibliographical references (leaves 83-87).
Abstracts in English and Chinese.
TABLE OF CONTENTS --- p.i
LIST OF FIGURES --- p.v
LIST OF TABLES --- p.vii
ABSTRACT --- p.viii
ACKNOWLEDGEMENT --- p.x
DECLARATION --- p.xi
ABBREVIATIONS --- p.xii
Chapter Chapter one --- Introduction
Chapter 1.1 --- Background --- p.1
Chapter 1.2 --- Polysaccharides isolated from Ganoderma Lucidum --- p.4
Chapter 1.3 --- Conventional methods for molecular weight (MW) determination of polysaccharides --- p.6
Chapter 1.3.1 --- Osmometry --- p.7
Chapter 1.3.2 --- Light Scattering --- p.8
Chapter 1.3.3 --- Intrinsic Viscosity --- p.8
Chapter 1.3.4 --- Size Exclusion Chromatography (SEC) --- p.9
Chapter 1.3.5 --- Mass Spectrometry --- p.10
Chapter 1.4 --- Matrix-assisted Laser Desorption / Ionization Mass Spectrometry --- p.12
Chapter 1.5 --- MALDI-TOF Mass Spectrometry of polysaccharides --- p.13
Chapter 1.6 --- Outline of project --- p.15
Chapter Chapter two --- Instrumental and experimental
Chapter 2.1 --- Instrumentation --- p.18
Chapter 2.1.1 --- Laser-based ion source --- p.18
Chapter 2.1.2 --- Time-of-flight (TOF) analyzer --- p.19
Chapter 2.1.3 --- Ion deflector --- p.23
Chapter 2.1.4 --- Detector and data acquisition system --- p.23
Chapter 2.2 --- Experimental --- p.26
Chapter 2.2.1 --- Isolation of water-soluble polysaccharides from Ganoderma Lucidum by water extraction --- p.26
Chapter 2.2.2 --- Isolation of water-soluble polysaccharides from Ganoderma Lucidum by dimethyl ssulfoxide (DMSO) extraction --- p.26
Chapter 2.2.3 --- Fractionation of water-soluble polysaccharides by Gel Permeation Chromatography (GPC) --- p.27
Chapter 2.2.4 --- Bradford protein assay --- p.28
Chapter 2.2.5 --- Phenol / sulfuric acid assay --- p.28
Chapter 2.2.6 --- Sample preparation in MS --- p.28
Chapter 2.2.7 --- Calibration of MALDI-TOF-MS --- p.29
Chapter 2.2.8 --- Data analysis --- p.29
Chapter Chapter three --- Extraction and purification of water-soluble polysaccharides from Ganoderma Lucidum
Chapter 3.1 --- Introduction --- p.30
Chapter 3.2 --- Experimental --- p.32
Chapter 3.2.1 --- Extraction efficiency --- p.32
Chapter 3.2.2 --- Dialysis --- p.32
Chapter 3.2.3 --- Signal suppression effect --- p.32
Chapter 3.2.4 --- Sevag method --- p.33
Chapter 3.2.5 --- Trichloroacetic acid (TCA) precipitation --- p.33
Chapter 3.2.6 --- Bradford Protein Assay --- p.34
Chapter 3.2.7 --- Phenol sulfuric acid assay --- p.34
Chapter 3.3 --- Results and discussion --- p.35
Chapter 3.3.1 --- Extraction efficiency --- p.35
Chapter 3.3.2 --- Purification of crude polysaccharides --- p.37
Chapter 3.3.3 --- Desalting --- p.38
Chapter 3.3.4 --- Deproteination --- p.40
Chapter 3.3.4.1 --- Monitoring of protein contents --- p.40
Chapter 3.3.4.2 --- Monitoring of carbohydrate contents --- p.45
Chapter 3.3.4.3 --- Deproteination using Sevag and TCA procipitation method --- p.47
Chapter 3.4 --- Conclusions --- p.54
Chapter Chapter four --- Evaluation of MW and MWD of water-soluble Polysaccharides extracted from Ganoderma Lucidum
Chapter 4.1 --- Introduction --- p.55
Chapter 4.2 --- Experimental --- p.58
Chapter 4.2.1 --- Aqueous DHB matrix --- p.58
Chapter 4.2.2 --- Aqueous DHB/NH4F matrix --- p.58
Chapter 4.2.3 --- Aqueous DHB matrix in TA solution --- p.58
Chapter 4.2.4 --- Aqueous DHB/NH4F matrix in TA solution --- p.59
Chapter 4.2.5 --- Aqueous DHB/NH4F matrix with sodium salt in TA solution --- p.59
Chapter 4.2.6 --- Aqueous DHB/NH4F matrix with potassium salt in TA solution --- p.59
Chapter 4.2.7 --- Fractionation of water-soluble polysaccharide extracted by DMSO by Gel Permeation Chromatography (GPC) --- p.59
Chapter 4.2.8 --- Ultra-violet absorption spectrometry (UV-VIS) --- p.60
Chapter 4.3 --- Results and discussion --- p.60
Chapter 4.3.1 --- Development of matrix and solvent system for water-soluble polysaccharides --- p.60
Chapter 4.3.2 --- MW and MWD of water-soluble polysaccharides extracted from Ganoderma Lucidum --- p.64
Chapter 4.3.2.1 --- Water extraction --- p.65
Chapter 4.3.2.2 --- DMSO extraction followed by water-back extraction --- p.70
Chapter 4.4 --- Conclusions --- p.79
Chapter Chapter five --- Concluding remarks --- p.82
References --- p.84

碩士
中原大學
醫學工程研究所
90
Ganoderma lucidum, has enjoyed a critically acclaimed reputation for its healing properties as a form of health food lies in none other than an extended encyclopedic testimonies, backed by documented evidence derived from recent research publications and clinical findings. Amongst a deluge of sensational Ganoderma lucidum studies of late, a majority of them have focused largely on examining its treatment properties, mycelium colony typing, improvement of means in culturing, and manipulation of culturing criteria. The study takes to developing a premise built upon reference archives with which to examine factors that may come to affect the liquid culture of Ganoderma lucidum, which entail validating the the cultivating temperature, rate of rotating the cultivation box for increasing soluble oxygen, the commencing pH rating, and how some other variables in the culturing process may come to affect to output, and in search of establishing an optimized operating criteria. The deign of the two-level factorial experiment lies in examining variables behind the culture solution, particularly of the individual yield and compound yield among the three critical factors of the commencing pH rating, metering of olive oil, and fungal metabolic additive with which to define the preliminary experimental objectives. Preliminary findings conclude that olive oil metering offers the highest individual yield, where E2 is measured at 708.3 ± 53.6, to far exceed the commencing pH rating, where E1 equals to 58.8 ± 53.6, or the fungal metabolic additive, where E3 equals to 88.8 ± 53.6, indicating that the presence of olive oil during the culturing process can indeed onerously excel the rate of mycelium culture. Noteworthy in the are of compound yield is how the simultaneous adaptation of olive oil and the metabolic addition, where E23 equals to 81.3 ± 53.6, would negatively impacted the results to lead to a decline in mycelium growth. Phase two of the experiment has the experiment’s factorial design extended to include five factors, which entail continuing with the previous three factors but adding two more factorial elements of temperature and rate of rotating the cultivation box. And the perspective of a fractional elemental experiment design has proven effective to conclude the individual yield, E1, of all elements with a reduction to the number of experiments required, and to examine the compound effect the element of olive oil metering comes to response to other elements, such as E12, E23, E24 and E25. Lastly, the experiment of factorial design takes to a regression analysis as the basis for design extrapolation and led to the conclusion an optimal model for the three critical cultivation control elements, namely the commencing pH rating, the metering of olive oil and Ganoderma lucidum metabolic additive, is at a commencing pH rating of 4.42, olive oil metering of 2.95 ml, Ganoderma lucidum metabolic additive at 1.16ml, cultivation temperature at 30℃. Celsius and rate of rotating the cultivation box at 100 rpm to conclude an optimized output of 1,614 mg.

碩士
國立臺灣大學
園藝暨景觀學系
103
Ganoderma lucidum, a medicinal fungi, has been used in Traditional Chinese Medicine (TCM) for its health promoting constituents, including polysaccharides, proteins, triterpenes, sterols, and fatty acids. However, G. lucidum contains various and complex bioactive compounds. It was difficult to distinguish the bioactivity of commercial G. lucidum products from specific component. Most products were claimed on the basis of their total sugar or crude polysaccharides contents, but immunomodulatory protein (LZ-8) and (1,3)-β-D-glucan, were not shown in the nutrition facts. In this study, we decided to examine whether the contents of total sugar or (1,3)-β-D-glucan could be used as the quality and functional indicators, and what would be the appropriate concentrates for the evaluation in product activities. First, we determined content of total sugar and content of (1,3)-β-D-glucan in commercial products with phenol-sulfuric acid method and fluorometric method. Our results showed that there were significant differences on the contents of total sugar and (1,3)-β-D-glucan in the eight commercial products. Samples with the same contends of total sugar or (1,3)-β-D-glucan had different capability to induce secreation of TNF-α and IL-6 in murine macrophages and RAW 264.7. Therefore, the quantity of total sugar and (1,3)-β-D-glucan might not be an appropriate functional indicators. Our results also revealed that the approach of diluting the commercial product solutions which were cultured with murine macrophages to 30 to 250 mg commercial product in 100 mL PBS could be used to distinguish bioactivities of commercial products. Further, the eight commercial products could not increase production of cytokine (IFN-γ, IL-2, and IL-4) by murine splenocytes. In addition, SDS-PAGE and Western blotting analysis of these samples showed that commercial G. lucidum products could rarely include immunomodulatory protein LZ-8. In the third part, mycelia from G. lucidum were lysed by ultrasonic reactor and analyzed for the content of LZ-8 by HPLC. LZ-8 peak have been observed as lysis solution filtered through glass fiber membranes and added 100 μg/mL standard LZ-8 to the solution. As shown above, we assumed that the component which interfered the HPLC analysis of LZ-8 should be polysaccharide. Sample solution was further treated with exo-(1,3)-β-D-glucanase and endo-(1,3)-β-D-glucanase in order to remove the potented interfering substance, but no LZ-8 peak have been observed. As a result, it was inferred that the interfering substance might be polysaccharides but not (1,3)-β-D-glucans. In conclusion, the content of total sugar, (1,3)-β-D-glucan, and LZ-8 could not be the functional indicators. The appropriate dosages of sample cultured with murine macrophages were 30 to 250 mg of commercial product in 100 mL PBS and could be used to assess product activities. In the third part, we inferred that polysaccharids might be the main factor to interfer the HPLC analysis of the free form of LZ-8.

碩士
國立宜蘭大學
生物資源學院碩士在職專班
103
In the health foods, Ganoderma is a familiar and favor product for consumes. Ganoderma has many bioactive functions, such as: antioxidant properties, immunomodulatory, hypoglycemic activity, hepatoprotective effects and other effects. Ganoderma has many active components such as polysaccharides and triterpenoids, which are widely used for research. The compositions such as: starch, protein, essential amino acids, free fatty acids and ash or the physical properties of cereals or legumes will be changed during fermentation. The objectives of this study were (1) to study the effect of Ganoderma lucidum solid-state fermentation on physicochemical properties of brown indica rice, (2) to develop rice milks using Ganoderma fermented rice, and to investigate the sensory characteristics and texture of the functional products. Indica rice was used as solid-state medium for 35 days Ganoderma lucidum solid-state fermentation, and polysaccharides production reached a maximum after 28 days cultivation, while crude triterpenoids would significantly increased with fermentation time. Indica rice after Ganoderma solid-state fermentation increased crude ash, crude protein, sugar, essential amino acids content, but carbohydrate content decreased. Indica rice after Ganoderma solid-state fermentation significantly decreased and was pH value, the color deepen as fermentation time increased. Unfermented brown indica rice flour had a lower peak viscosity, breakdown and higher setback, and Ganoderma solid-state and fermented brown rice powder had no clear peak viscosity, holding strength, final viscosity, breakdown, setback. The physicochemical properties of commercial rice milks were analyzed. The rice milk with new flavors Ganoderma lucidum rice milk, Ganoderma lucidum almond rice milk, Ganoderma lucidum sesame rice milk were developed and organic brown rice was as a commercial product. The sugar contents of the rice milks were between 6.1 -11.1 °Brix, and pH values of the rice milk were between 5.93-6.84. Analysis of L*, a*, b* values converted into whiteness between 29.43-65.95, orginal rice milk highest reach 65.95 color subdued, while Ganoderma sesame rice milk was the darkest color dim. Consistency index of organic brown rice milk was the highest index, while of consistency index Ganoderma almond rice milk was the lowest. Questionnaire survey of health food consumption patterns was drinks, which were 47.11 % with the highest value. The nine point hedonic scale test, Ganoderma lucidum almond rice milk and Ganoderma lucidum sesame rice milk have higher scores tasting flavor Ganoderma rice milk, orginal rice milk and organic brown rice, so mixing with other ingredients of Ganoderma rice milk development, helpfully provided consumers with more choice for Ganoderma related products, and to improve commercial Ganoderma consumption patterns.

碩士
國立嘉義大學
生物農業科技學系碩士班
98
Dye-type peroxidase (DyP) is a novel peroxidase gene family distinct from the plant peroxidase superfamily. This peroxidase family catalyzes not only typical peroxidase substrates, but also hydroxyl-free anthraquinone pigments. Ganoderma lucidum is a member of white-rot fungi characterized by their ability of lignin degradation. This fungal species also possesses good ability to tolerate heavy metal toxicity. In our previous study, a Cadmium-inducible gene isolated from Ganoderma lucidum exhibited high degree of similarity to the DyP-type peroxidase family in its predicted amino acid sequence (Chuang et al., 2009). To further characterize the physiological role of this DyP in fungal development, we first isolated its full length cDNA by the RLM-RACE method. The coding sequence of the Ganoderma’s cDNA, designated as GlDyP, exhibited 56-63% similarity in its predicted amino acid sequence to DyP genes in various fungal species. The amino acid sequence of GlDyP contained conserved amino acid residues found in other members of the DyP peroxidase, and residues corresponding for the heme binding sites. The phylogenetic analysis revealed that the amino acid sequence of GlDyP was closer related to that of DyP in Coprinopsis cinerea and Laccaria bicolor. The Northern blot analysis indicated that the GlDyP expression showed early induction in response to H2O2, Cd2+, Cu2+, and high osmotic stress. Addition of Zn2+ did not solicit the GlDyP expression. However, high salt condition suppressed this gene expression. In addition to responding to agents included in the culture medium, high culture temperature also induced the GlDyP expression in the later time course stage. The time course of inducing the GlDyP expression was well correlated with mycelium growth activity in response to various abiotic stress environments that the growth of Gandomera’s mycelium exhibited tolerance toward heavy metal of Cd2+, Cu2+, but not to Zn2+, and also toward osmotic stress. In contrast, the mycelium growth was hypersensitive to the addition of sodium chloride. In summary, our results indicated that GlDyP might play a role of detoxification in fungal cells encountering abiotic stress.

by Fu Sai-chuen.
Thesis (M.Phil.)--Chinese University of Hong Kong, 1994.
Includes bibliographical references (leaves 102-111).
Acknowledgements
List of Abbreviation
Abstract
Prologue --- p.1
Chapter
Chapter 1. --- Introduction --- p.2
Chapter 1.1 --- Historical background of Ling Zhi (靈芝） --- p.4
Chapter 1.2 --- Biological description of Ling Zhi (靈芝） --- p.6
Chapter 1.3 --- Chemical composition of Ganoderma lucidum --- p.6
Chapter 1.3.1 --- The triterpenoids --- p.7
Chapter 1.3.2 --- Matrix polysaccharides --- p.8
Chapter 1.4 --- Medicinal properties and biological activities --- p.9
Chapter 1.4.1 --- Medicinal properties --- p.9
Chapter 1.4.2 --- Biological activities --- p.10
Chapter 1.4.2.1 --- Anti-tumour activity --- p.10
Chapter 1.4.2.2 --- Enhancement of protein and nucleic acid synthesis --- p.11
Chapter 1.4.2.3 --- Effects on nervous system --- p.11
Chapter 1.4.2.4 --- Effects on respiratory system --- p.12
Chapter 1.4.2.5 --- Effects on cardiovascular system --- p.12
Chapter 1.4.2.6 --- Effects on immune system --- p.14
Chapter 1.4.2.7 --- Hepatoprotection and detoxicant actions --- p.14
Chapter 1.4.3 --- Summary --- p.15
Chapter 1.5 --- The relationship between antioxidant defense/immune system and disease --- p.15
Chapter 2. --- Materials and Methods --- p.20
Chapter 2.1 --- Cell culture reagents
Chapter 2.1.1 --- Preparation of RPMI-1640 medium --- p.20
Chapter 2.1.2 --- Preparation of fetal calf serum (FCS) --- p.20
Chapter 2.1.3 --- Preparation of phosphate-buffered saline (PBS) --- p.20
Chapter 2.1.4 --- Mitogens and test samples --- p.21
Chapter 2.1.5 --- Trypan blue exclusion test --- p.21
Chapter 2.1.6 --- Liquid scintillation counting --- p.21
Chapter 2.2 --- Animal care --- p.22
Chapter 2.3 --- Fractionation of Ganoderma lucidum extracts --- p.22
Chapter 2.4 --- Carbon tetrachloride (CC14)- induced hepatotoxicity in mice --- p.23
Chapter 2.4.1 --- Pretreatment scheme and CCl4 treatment --- p.23
Chapter 2.4.2 --- Preparation of liver homogenates --- p.23
Chapter 2.4.3 --- Biochemical assays for assessing hepatotoxicity --- p.25
Chapter 2.4.3.1 --- Determination of plasma alanine aminotransferase (ALT) --- p.25
Chapter 2.4.3.2 --- Determination of tissue glutathione (GSH) content --- p.25
Chapter 2.4.3.3 --- Depletion of tissue GSH in liver homogenates by tert- butylhydroperoxide (tBHP) --- p.26
Chapter 2.4.3.4 --- Determination of tissue malondialdehyde (MDA) content --- p.26
Chapter 2.5 --- Preparation of murine hepatocytes --- p.27
Chapter 2.5.1 --- Liver perfusion --- p.27
Chapter 2.5.2 --- Isolation of murine hepatocytes --- p.29
Chapter 2.5.3 --- Incubation protocol --- p.29
Chapter 2.5.4 --- Determination of cellular glutathione by HPLC method --- p.30
Chapter 2.6 --- Preparation of splenocytes and lymphocytes --- p.31
Chapter 2.6.1 --- Isolation of murine splenocytes --- p.31
Chapter 2.6.2 --- Mitogenic assay by measuring 3H-thymidine uptake --- p.31
Chapter 2.6.3 --- B cell- and T cell- enrichment in splenocyte preparation --- p.32
Chapter 2.6.4 --- Isolation of human peripheral blood lymphocytes (HPBL) from cord blood --- p.34
Chapter 2.7 --- Chromatographic methods --- p.34
Chapter 2.7.1 --- Fast Protein Liquid Chromatography (FPLC) with Superose 12 column --- p.34
Chapter 2.7.2 --- Removal of lipopolysaccharide (LPS) with Detoxi Gel --- p.35
Chapter 2.7.3 --- Desalting by Sephadex G-25 --- p.35
Chapter 2.7.4 --- Determination of FDNB-conjugated GSH and GSSG by HPLC reverse phase chromatography --- p.36
Chapter 2.8 --- Measurement of in vivo antibody production --- p.39
Chapter 2.8.1 --- Immunization and pretreatment scheme --- p.39
Chapter 2.8.2 --- Direct plaque assay --- p.39
Chapter 2.9 --- Biochemical analysis --- p.39
Chapter 2.9.1 --- Determination of protein content --- p.40
Chapter 2.9.2 --- Determination of hexose content --- p.40
Chapter 2.9.3 --- Determination of uronic acid content --- p.40
Chapter 2.9.4 --- Determination of sulphate content --- p.41
Chapter 2.9.5 --- Determination of hexosamine content --- p.42
Chapter 2.10 --- Statistical analysis --- p.42
Chapter 3. --- Results --- p.44
Chapter 3.1 --- Extraction and fractionation of Ganoderma lucidum --- p.44
Chapter 3.2 --- Hepatoprotective effect of Ganoderma lucidum fractions aganist CCl4-induced hepatotoxicity --- p.47
Chapter 3.2.1 --- Hepatoprotection of Ganoderma lucidum fractions --- p.47
Chapter 3.2.2 --- Effect of Ganoderma lucidum fraction pretreatment on hepatic malondialdehyde (MDA) level in CCl4-treated mice --- p.47
Chapter 3.2.3 --- Effect of Ganoderma lucidum fraction pretreatment on hepatic glutathione (GSH) level in CCl4-treated mice --- p.51
Chapter 3.3 --- Effect of GL2 on isolated hepatocytes --- p.51
Chapter 3.3.1 --- Protection against phorone-induced hepatocytotoxicity --- p.51
Chapter 3.3.2 --- Protection against menadione-induced hepatocytotoxicity --- p.55
Chapter 3.4 --- Immunomodulatory effect of Ganoderma lucidum fractions --- p.55
Chapter 3.4.1 --- Discovery of a mitogenic principle from Ganoderma lucidum fractions --- p.55
Chapter 3.4.2 --- B cell-specific mitogenic activity --- p.58
Chapter 3.4.3 --- Mitogenic activity in Ganoderma lucidum distinguishable from possible LPS contamination --- p.65
Chapter 3.4.3.1 --- The use of polymyxin B sulphate --- p.65
Chapter 3.4.3.2 --- The use of Detoxi Gel --- p.69
Chapter 3.4.3.3 --- The combined use of polymyxin B sulphate and Detoxi Gel --- p.69
Chapter 3.5 --- Enhancement of humoral response to sheep red blood cells (SRBC) by the mitogenic GL2 fraction --- p.72
Chapter 3.6 --- Physical-chemical characterization of GL2 fraction --- p.76
Chapter 3.6.1 --- Estimation of molecular size by FPLC size exclusion chromatography with Superose 12 column --- p.76
Chapter 3.6.2 --- Sugar composition of GL2 fraction --- p.76
Chapter 3.6.3 --- Comparison of elution profiles in Superose12 column with respect to mitogenic activity and sugar composition --- p.80
Chapter 4. --- Discussions --- p.83
Chapter 4.1 --- Enhancement of antioxidant status by Ganoderma lucidum fractions --- p.83
Chapter 4.2 --- The significance of enhancement of GSH status --- p.89
Chapter 4.3 --- Immunomodulatory actions of Ganoderma lucidum --- p.95
Conclusions --- p.99
Appendix --- p.100
Reference --- p.102

Sun Baizhong.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2004.
Includes bibliographical references (leaves 75-79).
Abstracts in English and Chinese.
ABSTRACT --- p.i
ABSTRACT (IN CHINESE) --- p.ii
TABLE OF CONTENTS --- p.iii
LIST OF FIGURES --- p.v
LIST OF TABLES --- p.vii
ABBREVIATIONS --- p.vi
ACKNOWLEDGEMENT --- p.ix
DECLARATION --- p.x
Chapter CHAPTER ONE --- INTRODUCTION
Chapter 1.1 --- Polysaccharides --- p.1
Chapter 1.2 --- Pharmacological importance of polysaccharides --- p.2
Chapter 1.3 --- Polysaccharides from Ganoderma Lucidum --- p.3
Chapter 1.4 --- Characterization of polysaccharides --- p.3
Chapter 1.5 --- Matrix-assisted laser desorption / ionization --- p.5
Chapter 1.6 --- Matrix-assisted laser desorption / ionization of polysaccharides --- p.8
Chapter 1.7 --- Outline of the project --- p.10
Chapter CHAPTER TWO --- INSTRUMENTATION AND EXPERIMENTAL
Chapter 2.1 --- Time-of-flight mass spectrometry --- p.12
Chapter 2.1.1 --- Instrumentation --- p.15
Chapter 2.1.1.1 --- Laser system --- p.16
Chapter 2.1.1.2 --- Ion source --- p.16
Chapter 2.1.1.3 --- Reflector --- p.19
Chapter 2.1.1.4 --- Detector --- p.19
Chapter 2.1.1.5 --- Data acquisition and manipulation --- p.20
Chapter 2.2 --- Ultraviolet-visible spectrometer --- p.21
Chapter CHAPTER THREE --- OPTIMIZATION OF MALDI CONDITIONS FOR POLYSACCHARIDE ANALYSIS USING DMSO AS SOLVENT
Chapter 3.1 --- Introduction --- p.22
Chapter 3.2 --- Experimental --- p.25
Chapter 3.2.1 --- Selection of matrix materials --- p.26
Chapter 3.2.2 --- Selection of co-matrix materials --- p.26
Chapter 3.2.3 --- Ratios of matrix-to-analyte --- p.26
Chapter 3.2.4 --- Effect of sample drying temperature --- p.27
Chapter 3.2.5 --- Sample loading methods --- p.27
Chapter 3.3 --- Results and discussion --- p.27
Chapter 3.3.1 --- Selection of matrix materials --- p.28
Chapter 3.3.2 --- Selection of co-matrix materials --- p.30
Chapter 3.3.3 --- Effect of matrix-to-co-matrix ratio --- p.32
Chapter 3.3.4 --- Effect of sample drying temperature --- p.37
Chapter 3.3.5 --- Effect of matrix-to-analyte ratio --- p.39
Chapter 3.3.6 --- Evaluation of sample preparation protocols --- p.43
Chapter 3.4 --- Conclusion --- p.46
Chapter CHAPTER FOUR --- MOLECULAR AND MOLECULAR WEIGHT DISTRIBUTION OF WATER-INSOLUBLE POLYSACCHARIDES FROM GANODERMA LUCIDUM
Chapter 4.1 --- Introduction --- p.47
Chapter 4.2 --- Experimental --- p.48
Chapter 4.2.1 --- Extraction of water-insoluble polysaccharides from Ganoderma Lucidum --- p.48
Chapter 4.2.2 --- Gel permeation chromatography of water-insoluble polysaccharides --- p.49
Chapter 4.2.3 --- Phenol sulfuric acid assay --- p.51
Chapter 4.2.4 --- MALDI-TOF analysis --- p.51
Chapter 4.2.5 --- Bradford assay --- p.52
Chapter 4.3 --- Results and discussion --- p.52
Chapter 4.3.1 --- Extraction of water-insoluble polysaccharides from Ganoderma Lucidum --- p.52
Chapter 4.3.2 --- Fractionation of water-insoluble polysaccharides using gel permeation chromatography (GPC) --- p.54
Chapter 4.3.3 --- MALDI-TOF analysis of fractionated polysaccharides --- p.57
Chapter 4.3.4 --- Carbohydrate content analysis of fractionated polysaccharides --- p.62
Chapter 4.3.5 --- MW and MWD of water-insoluble polysaccharides extracted from Ganoderma Lucidum --- p.69
Chapter 4.4 --- Conclusion --- p.71
Chapter CHAPTER FIVE --- CONCLUDING REMARKS
Chapter 5.1 --- Conclusion --- p.73
REFERENCES --- p.76
APPENDIX --- p.81
Appendix A Reaction scheme of carbohydrate with phenol in phenol-sulfuric acid assay --- p.81

博士
東海大學
化學工程學系
94
Previous works have shown that in many filamentous fungi fermentations the morphology of the fungi influence both biomass and metabolites. The formation of large pellets is the main characteristic of the mycelium submerged culture of G. lucidum. In this study the dynamic change of mycelium aggregation process was investigated in 5L fermentor. A novel scheme using intermittent agitation could maintain a higher level of mfc（mycelium fragment concentration） for more than 24 hours and enhanced the biomass concentration to the level of 2.28 g/L. By adding the polymers to the fermentation medium, the growth form of the G. lucidum was changed from large glob to small pellets, 0.2% (wt/vol) agar was found to be the optimum concentrations. The biomass concentration with 0.2% (w/v) agar was 2.52g/L, compared to 1.54g/L of the control. Mechanical shear force could reduce the pellet size, but also damaged the mycelium at the same time. Using intermittent homogenizer could change the morphology of G. lucidum from pelleted form to filamentous form and also promoted the biomass yield to 1.14g/L day-1.

碩士
國立宜蘭大學
食品科學系碩士班
99
Trivalent chromium (Cr3+) has been applied in type II diabetes to decrease blood glucose level and improve glucose tolerance. Ganoderma lucidum can modulate blood glucose in animals. The objectives of this study were to develop chromium enriched Ganoderma lucidum fermented product (ECGL) and evaluate its modulating function of glycemia. The results showed that the addition 100 ppm Cr3+ in liquid medium had higher biomass growth (1.78 g/L). Submerged fermentation of Ganoderma lucidum with 150 mL 5% cereal medium and 100 ppm Cr3+ in a 500 mL flask has the highest biomass 3.01 g/L and organic chromium 40.1 mg/L production after 7-days cultivation. The organic chromium of Ganoderma lucidum is a primary metabolite and 92% of organic chromium belongs to extracellular products. The oxygen transfer number (KLa) in 5-L air-lift and stirred fermentor with 5% cereal medium were analyzed. The KLa is between in air-lift fermentor even aeration was increase and the KLa is clearly improve from 11.3 to 111.1 hr-1 with same aeration when impeller rate increase. The operation at 300 rpm with KLa = 88.3 (hr-1) had the highest biomass (19.8 g/L) than the orther impeller rate operations. The Ganoderma lucidum fermentation in a medium contening 5% cereal and 100 ppm Cr3+ operating at 300 rpm impeller and 1 vvm aeration had the highest biomss (10.69 g/L) and organic chromium production (64.1 mg/L) after 7-days cultivation. The maximum organic chromium was after 14-days Ganoderma solid-state fermentation with wheat medium. The hightest organic chromium conversion (54.7%) was used as medium contening 200 ppm Cr3+. The fermentation product contening organic chromium 109.4 mg/kg and then use to oral feed STZ-mice after hot water extraction. The dose of extract solution was 75 mg ECGL/kg day for 2-week had clear decrease plasma glucose after a meal.