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Genetic, lipidic and proteomic characterization of an arachidonic acid producing fungus, Mortierella alpinaHo, Sze-yuen. January 2008 (has links)
Thesis (Ph. D.)--University of Hong Kong, 2009. / Includes bibliographical references (leaves 141-163) Also available in print.
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Genetic, lipidic and proteomic characterization of an arachidonic acidproducing fungus, Mortierella alpinaHo, Sze-yuen., 何思遠. January 2008 (has links)
published_or_final_version / Biological Sciences / Doctoral / Doctor of Philosophy
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Biochemical analysis and molecular breeding of oleaginous microorganisms for ω3 polyunsaturated fatty acid production / ω3高度不飽和脂肪酸生産のための油糧微生物の生化学的解析ならびに分子育種Okuda, Tomoyo 24 March 2014 (has links)
京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第18343号 / 農博第2068号 / 新制||農||1024(附属図書館) / 学位論文||H26||N4850(農学部図書室) / 31201 / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 小川 順, 教授 植田 充美, 教授 栗原 達夫 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM
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PRODUÇÃO DE ÁCIDOS GRAXOS POLI-INSATURADOS POR FERMENTAÇÃO SUBMERSA / POLYUNSATURATED FATTY ACIDS PRODUCTION BY SUBMERGED FERMENTATIONSomacal, Simara 17 December 2015 (has links)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The objective of this work was the polyunsaturated fatty acids (PUFAs)
production by submerged fermentation from the fungus Mortierella isabellina. A
Plackett Burman experimental design was designed to evaluate the best
conditions for PUFAs production in shake flasks. Three different components of
the culture medium (yeast extract, peptone, and sucrose) and three
fermentation parameters (agitation, temperature and pH) were
evaluated. Analyzing the results of experimental design new assays were
performed with increasing concentration of yeast extract. Fermentations were
conducted in Erlenmeyer flasks with 10% (v/v) inoculums for 120h,
micronutrients ((NH4)2SO4, FeSO4.7H2O, MnSO4.H2O, MgSO4.7H2O) were
fixed. The scale up using STR bioreactor with 2.5 L of culture medium and 10%
(v/v) inoculums for 120h was defined through better results of Plackett
Burman.. A 2² central composite design was set to evaluate the effect of
agitation and aeration in fungal biomass production, lipids content and fatty
acids profile. The lipid was extracted from dried biomass and the fatty acids
were analyzed by gas chromatography. The results of the fermentation in shake
flasks showed that only yeast extract had a significant effect (p <0.1) in PUFAs
production and this effect was positive. The highest percentage of PUFAs
produced (28.17%) was obtained in higher yeast extract concentration (3.75 g /
L). Agitation and aeration did not had a significance effect (p <0.1) in the
evaluated range, in bioreactor on PUFAs and lipids production. However,
agitation had a significant positive effect on biomass production.The presence
of fatty acids of family n-3 (α-linolenic acid, eicosapentaenoic acid and
docosahexaenoic acid) and n-6 families (linoleic acid and γ-linolenic acid) were
observed. Nitrogen source is directly related to increased PUFAs
production. The variables agitation and aeration, in the evaluated range, did not
influence the PUFAs production. The Mortierella isabellina produced fatty acids
n-3 and n-6 family, but additional studies are needed to
optimize this production. / O objetivo deste trabalho foi produzir ácidos graxos poli-insaturados (PUFAs)
por fermentação submersa a partir do fungo Mortierella isabellina. Um
planejamento experimental Plackett Burman foi delineado para avaliar as
melhores condições para produção de PUFAS em frascos agitados. Foram
avaliados três diferentes componentes do meio de cultura (extrato de levedura,
peptona e sacarose) e três parâmetros de fermentação (agitação, temperatura
e pH). Através dos resultados obtidos no planejamento experimental, novos
ensaios foram definidos com o aumento da concentração de extrato de
levedura. As fermentações foram realizadas em frascos Erlenmeyers com 10%
de inoculo (v/v) durante 120h e a composição de micronutrientes ((NH4)2SO4,
FeSO4.7H2O, MnSO4.H2O, MgSO4. 7H2O) foi mantida fixa. Na melhor condição
definida em frascos agitados foi realizada a ampliação da fermentação para
biorreator STR, com 2,5 L de meio de cultura e 10 % de inoculo (v/v) durante
120h. Um delineamento composto central 2² foi definido para avaliar o efeito
das variáveis agitação e aeração na produção de biomassa fúngica, lipídeos e
no perfil dos ácidos graxos. A fração lipídica foi extraída a partir da biomassa
seca da fermentação e os ácidos graxos foram analisados por cromatografia
em fase gasosa. Os resultados da fermentação em frascos agitados
demonstraram que somente a variável extrato de levedura teve efeito
significativo (p<0,1) na produção de PUFAs sendo este efeito positivo. O maior
percentual de PUFAs produzidos (28,17%) foi obtido na maior concentração de
extrato de levedura (3,75 g/L). Os resultados obtidos na fermentação em
biorreator demonstraram que as variáveis agitação e aeração não foram
estatisticamente significativas (p<0,1) para o acúmulo de PUFAs e lipídeos
dentro das faixas avaliadas. Entretanto a agitação teve efeito significativo
positivo na produção de biomassa. Foi observada a presença dos ácidos
graxos da família n-3 (ácido α-linolênico, ácido eicosapentaenóico e ácido
docosaexaenóico) e da família n-6 (ácido linoléico e o ácido γ-linolênico). Em
vista dos resultados obtidos, pode-se concluir que a fonte de nitrogênio esta
diretamente relacionada com o aumento da produção de PUFAs. As variáveis
agitação e aeração nas faixas avaliadas não influenciaram no acúmulo de
PUFAs. A Mortierella isabellina produziu ácidos graxos da família n-3 e n-6,
entretanto estudos adicionais devem ser realizados para otimizar esta
produção.
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Biotransformação in vitro de novos candidatos a protótipos de fármacos antitumorais N-fenilpirazóis / In vitro biotransformation of novel N-phenylpyrazole antitumor drug prototypeAraujo, Kelly Carolina Frauzino 28 February 2014 (has links)
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Previous issue date: 2014-02-28 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / The "Microbial Models of Mammalian Metabolism" represents an alternative to use of animals on metabolism studies. Introduced in the 70s this model, also called biotranformation, has several advantages for their application as low cost, reduction of animals for experimentation and and higher number and range of metabolites produced. 1-(4-((1-(4-chlorophenyl)-1H-pyrazol-4-yl)methyl)piperazin-1-yl)ethanone (LQFM030) and 4-((1-(4-chlorophenyl)-1H-pyrazol-4-il)methyl)piperazin-1-ethyl carboxylate (LQFM018) compounds were synthesized by molecular simplification of a series of compounds with chronic myeloid leukemia antiproliferative activity already described, the Nutlins prototypes. This study aims to produce a probable human metabolites of LQFM 030 and LQFM 018 by microbial biotranformation with filamentous fungi. To do so, analytical methodologies were developed by thin layer chromatography and high performance liquid chromatography in order to monitor metabolites production. After perform a screening of tem microorganisms Mortierella isabellina NRRL 1757 strain was selected to obtain metabolites on a larger scale. Incubations were carried out with 100 mL of glucose culture medium in each flask . At the end of incubation (96 h) extraction and purification of possible metabolites was performed . In an independent assay with LQFM 030, ketoconazole (10, 20 and 30 mg) was added to inhibit P450 cytochrome . In another test 1 mL of ethanol was added every 24 hours to induce cytochrome totaling 96 hours of incubation. To evaluate the best time to finalize incubation , an experiment was conducted in 168 hours with different concentrations of the substrate, 0.25 mg / mL and 50 mg / mL. A possible role of CYP 3A was evidenced by the inhibition caused by ketoconazole addition, that can inhibit the formation of N-oxide metabolite in LQFM 030 biotransformation. Ethanol addition does not induce LQFM 030-N-oxide production, but was able to induce other metabolites formation. The best time to end LQFM 030 incubation was defined as 168 hours and 0.25 mg/mL concentration. / O “Modelo Microbiano do Metabolismo Animal”, pode representar uma alternativa ao uso de animais nos estudos de metabolismo de novos candidatos a fármacos. Introduzido nos anos 70, este modelo, também denominado de biotransformação, apresenta várias vantagens para a sua aplicação como o baixo custo, a redução da utilização de animais de experimentação e maior quantidade e variedade de metabólitos produzidos. Os compostos 1-(4-((1-(4-clorofenil)-1H-pirazol-4-il)metil)piperazin-1-il)etanona (LQFM 030) e 4-((1-(4-clorofenil)-1H-pirazol-4-il)metil)piperazin-1-carboxilato de etila (LQFM 018) foram sintetizados a partir da simplificação molecular de uma série de compostos com atividade antiproliferativa frente à leucemia mielóide crônica já descrita, os protótipos Nutlins. O presente estudo objetivou a produção de prováveis metabólitos humanos do LQFM 030 e LQFM 018 através de biotransformação microbiana com fungos filamentosos. Para isso, foram desenvolvidas metodologias analíticas por cromatografia em camada delgada e cromatografia líquida de alta eficiência, a fim de se monitorar a produção dos metabólitos. Após seleção dentre dez microrganismos a cepa Mortierella isabellina NRRL 1757 foi escolhida para obtenção de metabólitos em maior escala. A incubação foi realizada com 100 mL de meio de cultura de glicose, em cada Erlenmeyer. Ao término da incubação (96 horas) foi realizada a extração e purificação dos possíveis metabólitos. Em um ensaio independente foi adicionado cetoconazol (10, 20 e 30 mg) ao meio contendo LQFM 030 com o objetivo de inibir o citocromo P450. Em outra etapa foi adicionado 1 mL de etanol a cada 24 horas para induzir o citocromo, totalizando 96 horas de incubação. Para avaliar a melhor cinética reacional, um experimento foi conduzido em 168 horas, com diferentes concentrações do substrato, 0,25 mg/mL e 50 mg/mL. A biotransformação do LQFM 030 e do LQFM 018 produziu seus respectivos N-óxidos, caracterizados por RMN 1H e 13C e EM. Uma provável participação da CYP 3A foi evidenciada através da inibição causada pela adição de cetoconazol capaz de inibir a formação do metabólito N-óxido na biotransformação do LQFM 030. A adição de etanol não induziu a produção do LQFM 030-N-óxido, contudo foi capaz de induzir a formação de outros metabólitos. O melhor tempo para término da incubação do LQFM 030 foi definido como 168 hrs e 0,25 mg/mL de concentração.
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The production of the highly unsaturated fatty acid eicosapentaenoic acid by fungal solid state fermentationJacobs, Annali 03 1900 (has links)
Thesis (MSc (Microbiology))--University of Stellenbosch, 2010. / ENGLISH ABSTRACT: Long chain omega-3 fatty acids such as eicosapentaenoic acid (EPA) are
essential for the regulation of critical biological functions in humans and
other mammals. Fish oil as the main dietary source of EPA holds several
disadvantages and alternative sources and production processes such as
microbial fermentation are increasingly being investigated.
Therefore the aim of the first part of this study was to evaluate brewers’
spent grain (BSG) as substrate for the production of EPA by solid state
fermentation with 29 fungal strains representing different Mortierella
species. The effect of a 10% (w/w) linseed oil (LSO) supplement on EPA
production was also studied. Consequently, fungal inoculated BSG was
incubated at 22oC for three days to obtain optimal fungal growth, before
the incubation temperature was lowered to 16oC for the following eight
days. Cultures were then harvested and dried, followed by lipid extraction
and analyses using gas chromatography.
All the strains were found to produce EPA on BSG, while addition of the
LSO improved the EPA yield of most strains. The strains which produced
the highest levels of EPA on BSG supplemented with LSO were
Mortierella antarctica Mo 67 and Mortierella epicladia Mo 101, which
respectively produced 2.8 mg and 2.5 mg EPA per g of BSG.
During the second part of the study eight Mortierella strains were used to
study EPA production via solid state fermentation of sunflower press cake
(SPC). Similar culture conditions and analytical methods were used as in
the first part of the study. The effect of supplementing the SPC substrate
with 10% (w/w) LSO was studied with regard to the supplement’s impact
on EPA production and on the highly unsaturated fatty acid (HUFA) profile
of the fermented substrate. Addition of LSO improved EPA yield of most strains on SPC, leading to a
reduction in the average arachidonic acid (ARA):EPA ratio from 50.68 to
3.66. The ratio of HUFA to saturated and monoenoic fatty acids, was
increased significantly (t=5.75, p=0.05) by the addition of LSO, with higher
desaturation levels among the 20-carbon fatty acids. Addition of LSO also
had a positive effect (r = 0.9291, p = 0.001) on the relative amount of long
chain fatty acids (C≥20) produced. The strains which produced the highest
levels of EPA on SPC supplemented with LSO were Mortierella alpina Mo
46 and Mortierella basiparvispora Mo 88, which produced 6.4 mg and 5.8
mg EPA per g of sunflower press cake, respectively.
Fungi belonging to the genus Mortierella successfully converted LSO
supplemented agro-processing wastes, such as BSG and SPC, to
materials containing EPA, thereby adding value to these substrates. These
EPA-enriched waste substrates could eventually find applications as
animal or fish feed or as a source of EPA and other HUFA for the growing
omega-3 market in the neutraceutical and therapeutics industry. / AFRIKAANSE OPSOMMING: Langketting omega-3 vetsure soos eikosapentaenoë suur (EPS) is
noodsaaklik vir die regulasie van kritiese biologiese funksies in mense en
ander soogdiere. Visolie, die mees belangrike EPS-bron in die dieet, hou
verskeie nadele in en alternatiewe bronne sowel as produksie-prosesse,
soos mikrobiologiese fermentasie, word dus toenemend ondersoek.
Die doel van die eerste gedeelte van hierdie studie was dus om gebruikte
brouersgraan (GBG) te ëvalueer as ‘n substraat vir die produksie van EPS
deur soliede staat fermentasie met 29 fungus isolate wat verskillende
Mortierella spesies verteenwoordig. Die uitwerking van byvoeging van
10% (m/m) lynsaadolie (LSO) op EPS-produksie is ook bepaal. Gevolglik
is fungus-geïnokuleerde GBG vir drie dae by 22oC geïnkubeer om
optimale fungusgroei te verkry, waarna die inkubasie temperatuur verlaag
is na 16oC vir die volgende agt dae. Kulture is hierna ge-oes en gedroog,
gevolg deur lipied ekstraksie en analise met behulp van
gaschromatografie.
Al die isolate het EPS geproduseer op die GBG substraat, terwyl
byvoeging van LSO die EPS-opbrengs van die meeste isolate verbeter
het. Die isolate wat die hoogste vlakke van EPS op GBG wat met LSO
verryk is, geproduseer het, was Mortierella antarctica Mo 67 en Mortierella
epicladia Mo 101, wat onderskeidelik 2.8 mg en 2.5 mg EPS per g GBG
geproduseer het.
Tydens die tweede gedeelte van die studie is agt Mortierella isolate
gebruik om die produksie van EPS deur soliede staat fermentasie van
sonneblom perskoek (SPK) te ondersoek. Kultuurtoestande en analitiese
metodes soortgelyk aan die eerste gedeelte van die studie is gebruik. Die
uitwerking van byvoeging van 10% LSO tot die SPK substraat is
ondersoek met betrekking tot die impak van die byvoeging op EPS produksie asook op die profiel van hoogs onversadigde vetsure (HOVS)
van die gefermenteerde substraat.
Die byvoeging van LSO tot SPK het die EPS opbrengs van meeste isolate
verbeter en het tot ‘n verlaging in die gemiddelde arachidoonsuur
(ARS):EPS verhouding vanaf 50.69 tot 3.66 gelei. Die verhouding van
HOVS tot versadigde en mono-onversadigde vetsure, is betekenisvol
(t=5.75, p=0.05) verhoog deur die byvoeging van LSO, met hoër vlakke
van onversadigheid onder die 20-koolstof vetsure. Byvoeging van LSO het
ook ‘n positiewe uitwerking (r = 0.9291, p = 0.001) op die relatiewe aantal
langketting vetsure (C≥20) gehad. Die isolate wat die hoogste vlakke van
EPS geproduseer het op LSO-verrykte SPK, was Mortierella alpina Mo 46
en Mortierella basiparvispora Mo 88, wat onderskeidelik 6.4 mg en 5.8 mg
EPS per g SPK geproduseer het.
Fungi wat aan die genus Mortierella behoort, het LSO-verrykte agroprosesserings
afvalprodukte, soos GBG en SPK, suksesvol omgeskakel
na materiale wat EPS bevat, en sodoende waarde toegevoeg aan hierdie
substrate. Die EPS-verrykte afvalsubstrate kan uiteindelik toepassings
vind as diere- of visvoer of as bron van EPS of ander HOVS vir die
groeiende omega-3 mark in die neutraseutiese en terapeutiese industrie.
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Biochemical analysis and genetic engineering of oleaginous fungi for the production of eicosapentaenoic acid and free fatty acid derivatives / エイコサペンタエン酸と遊離脂肪酸誘導体生産のための油糧糸状菌の生化学的解析と遺伝子工学Brian, King Himm Mo 23 March 2021 (has links)
京都大学 / 新制・課程博士 / 博士(農学) / 甲第23253号 / 農博第2460号 / 新制||農||1085(附属図書館) / 学位論文||R3||N5343(農学部図書室) / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 小川 順, 教授 阪井 康能, 教授 栗原 達夫 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM
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有用油脂生産のための油糧糸状菌の代謝解析と効率的遺伝子ターゲティングシステムの構築 / Metabolic analysis and development of efficient gene-targeting systems in oleaginous fungi for useful lipid production菊川, 寛史 23 March 2015 (has links)
Kyoto University (京都大学) / 0048 / 新制・課程博士 / 博士(農学) / 甲第19047号 / 農博第2125号 / 新制||農||1032 / 31998 / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 小川 順, 教授 喜多 恵子, 教授 栗原 達夫 / 学位規則第4条第1項該当
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Metabolic analysis and development of efficient gene-targeting systems in oleaginous fungi for useful lipid production / 有用油脂生産のための油糧糸状菌の代謝解析と効率的遺伝子ターゲティングシステムの構築Kikukawa, Hiroshi 23 March 2015 (has links)
京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第19047号 / 農博第2125号 / 新制||農||1032(附属図書館) / 学位論文||H27||N4929(農学部図書室) / 31998 / 京都大学大学院農学研究科応用生命科学専攻 / (主査)教授 小川 順, 教授 喜多 恵子, 教授 栗原 達夫 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM
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