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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Molecular mechanisms of action and activation of lipoprotein lipase

Vainio, Petri. January 1985 (has links)
Thesis (doctoral)--University of Helsinki, 1986. / Includes bibliographical references (p. 31-38).
2

Functional characterisation of a novel ferulic acid esterase from Malawian hot spring metagenome

Ngobeni, Rhulani January 2011 (has links)
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mso-tstyle-colband-size:0 / mso-style-noshow:yes / mso-style-priority:99 / mso-style-parent:"" / mso-padding-alt:0cm 5.4pt 0cm 5.4pt / mso-para-margin-top:0cm / mso-para-margin-right:0cm / mso-para-margin-bottom:10.0pt / mso-para-margin-left:0cm / line-height:115% / mso-pagination:widow-orphan / font-size:11.0pt / font-family:"Calibri","sans-serif" / mso-ascii-font-family:Calibri / mso-ascii-theme-font:minor-latin / mso-hansi-font-family:Calibri / mso-hansi-theme-font:minor-latin / mso-bidi-font-family:"Times New Roman" / mso-bidi-theme-font:minor-bidi / mso-fareast-language:EN-US / } </style> <![endif]--> <p style="margin-bottom:0cm / margin-bottom:.0001pt / text-align: justify / line-height:normal / mso-layout-grid-align:none / text-autospace:none" class="MsoNormal"><span style="font-size:10.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / ">There has been a decline in the global fossil fuel reserves, due to an increasing demand for petroleum. Biofuels can be used as an alternative source of energy whereby biomass is converted to liquid fuels such as bioethanol. There is considerable interest in lipolytic enzymes because of their broad substrate range and for this purpose these enzymes have potential in a variety of biotechnological application. Lipolytic enzymes include esterases (E.C 3.1.1.1) and lipases (E.C3.1.1.3). Esterases preferentially hydrolyse short chain (&lt / C10) ester-containing molecules that are partly soluble in water, while lipases hydrolyse a broad range of substrates preferably water-insoluble fatty acyl molecules (&gt / C10). The aim of this study was to express, purify and characterise the lipolytic enzyme present on afosmid, Try 11, previously isolated from a metagenomic library of a Malawian hotspring, which conferred activity on tributyrin and ethyl ferulate. Bioinformatic analysisof the fosmid insert sequence predicted an open reading frame consisting of 951 bp,designated RHgene34, encoding a 317 amino acid protein with 41 % similarity to the</span><span style="font-size: 10.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / mso-fareast-font-family:ArialOOEnc">&alpha / /&beta / hydrolase fold-3 domain protein of </span><i><span style="font-size:10.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / ">Burkholderia </span></i><span style="font-size:10.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / ">sp. The RHgene34 protein </span><span style="font-size:10.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / ">contains conserved motifs of esterases/lipases, such as HGGG (residues 95-98),GxSxG (residues 167 - 171) and the putative catalytic triad composed of Ser157,Asp255 and His285. The gene was cloned and expressed in pET21a(+), and transformed into <i>Escherichia coli </i>Rosetta. <i>p</i>-Nitrophenol (<i>p</i>-Np) fatty acyl esters of different carbon chain lengths were used for kinetic characterisation of RHgene34. </span><span style="font-size:10.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / ">Kinetic analysis revealed that RHgene34 had a broad range activity on the <i>p</i>-Npesters, from C2 - C14. RHgene34 operates optimally at 45 &ordm / C, pH 9.0 and has a half life of 30 mins at 45 &ordm / C. This study demonstrates that functional screening combined with the sequence analysis is a useful approach for isolating novel enzymes from ametagenome.</span></p> <p style="margin-bottom:0cm / margin-bottom:.0001pt / text-align: justify / line-height:normal / mso-layout-grid-align:none / text-autospace:none" class="MsoNormal"><span style="font-size:12.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / "><br type="_moz" /> </span></p> <br /> <p style="margin-bottom:0cm / margin-bottom:.0001pt / text-align: justify / line-height:normal / mso-layout-grid-align:none / text-autospace:none" class="MsoNormal"><span style="font-size:12.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / "><br type="_moz" /> </span></p> <br />
3

Functional characterisation of a novel ferulic acid esterase from Malawian hot spring metagenome

Ngobeni, Rhulani January 2011 (has links)
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mso-tstyle-colband-size:0 / mso-style-noshow:yes / mso-style-priority:99 / mso-style-parent:"" / mso-padding-alt:0cm 5.4pt 0cm 5.4pt / mso-para-margin-top:0cm / mso-para-margin-right:0cm / mso-para-margin-bottom:10.0pt / mso-para-margin-left:0cm / line-height:115% / mso-pagination:widow-orphan / font-size:11.0pt / font-family:"Calibri","sans-serif" / mso-ascii-font-family:Calibri / mso-ascii-theme-font:minor-latin / mso-hansi-font-family:Calibri / mso-hansi-theme-font:minor-latin / mso-bidi-font-family:"Times New Roman" / mso-bidi-theme-font:minor-bidi / mso-fareast-language:EN-US / } </style> <![endif]--> <p style="margin-bottom:0cm / margin-bottom:.0001pt / text-align: justify / line-height:normal / mso-layout-grid-align:none / text-autospace:none" class="MsoNormal"><span style="font-size:10.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / ">There has been a decline in the global fossil fuel reserves, due to an increasing demand for petroleum. Biofuels can be used as an alternative source of energy whereby biomass is converted to liquid fuels such as bioethanol. There is considerable interest in lipolytic enzymes because of their broad substrate range and for this purpose these enzymes have potential in a variety of biotechnological application. Lipolytic enzymes include esterases (E.C 3.1.1.1) and lipases (E.C3.1.1.3). Esterases preferentially hydrolyse short chain (&lt / C10) ester-containing molecules that are partly soluble in water, while lipases hydrolyse a broad range of substrates preferably water-insoluble fatty acyl molecules (&gt / C10). The aim of this study was to express, purify and characterise the lipolytic enzyme present on afosmid, Try 11, previously isolated from a metagenomic library of a Malawian hotspring, which conferred activity on tributyrin and ethyl ferulate. Bioinformatic analysisof the fosmid insert sequence predicted an open reading frame consisting of 951 bp,designated RHgene34, encoding a 317 amino acid protein with 41 % similarity to the</span><span style="font-size: 10.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / mso-fareast-font-family:ArialOOEnc">&alpha / /&beta / hydrolase fold-3 domain protein of </span><i><span style="font-size:10.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / ">Burkholderia </span></i><span style="font-size:10.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / ">sp. The RHgene34 protein </span><span style="font-size:10.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / ">contains conserved motifs of esterases/lipases, such as HGGG (residues 95-98),GxSxG (residues 167 - 171) and the putative catalytic triad composed of Ser157,Asp255 and His285. The gene was cloned and expressed in pET21a(+), and transformed into <i>Escherichia coli </i>Rosetta. <i>p</i>-Nitrophenol (<i>p</i>-Np) fatty acyl esters of different carbon chain lengths were used for kinetic characterisation of RHgene34. </span><span style="font-size:10.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / ">Kinetic analysis revealed that RHgene34 had a broad range activity on the <i>p</i>-Npesters, from C2 - C14. RHgene34 operates optimally at 45 &ordm / C, pH 9.0 and has a half life of 30 mins at 45 &ordm / C. This study demonstrates that functional screening combined with the sequence analysis is a useful approach for isolating novel enzymes from ametagenome.</span></p> <p style="margin-bottom:0cm / margin-bottom:.0001pt / text-align: justify / line-height:normal / mso-layout-grid-align:none / text-autospace:none" class="MsoNormal"><span style="font-size:12.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / "><br type="_moz" /> </span></p> <br /> <p style="margin-bottom:0cm / margin-bottom:.0001pt / text-align: justify / line-height:normal / mso-layout-grid-align:none / text-autospace:none" class="MsoNormal"><span style="font-size:12.0pt / font-family:&quot / Arial&quot / ,&quot / sans-serif&quot / "><br type="_moz" /> </span></p> <br />
4

Functional characterisation of a novel ferulic acid esterase from malawian hot spring metagenome

Ngobeni, Rhulani January 2011 (has links)
>Magister Scientiae - MSc / There has been a decline in the global fossil fuel reserves, due to an increasing demand for petroleum. Biofuels can be used as an alternative source of energy whereby biomass is converted to liquid fuels such as bioethanol. There is considerable interest in lipolytic enzymes because of their broad substrate range and for this purpose these enzymes have potential in a variety of biotechnological application. Lipolytic enzymes include esterases (E.C 3.1.1.1) and lipases (E.C3.1.1.3). Esterases preferentially hydrolyse short chain (<C10) ester-containing molecules that are partly soluble in water, while lipases hydrolyse a broad range of substrates preferably water-insoluble fatty acyl molecules (>C10). The aim of this study was to express, purify and characterise the lipolytic enzyme present on afosmid, Try 11, previously isolated from a metagenomic library of a Malawian hotspring, which conferred activity on tributyrin and ethyl ferulate. Bioinformatic analysisof the fosmid insert sequence predicted an open reading frame consisting of 951 bp,designated RHgene34, encoding a 317 amino acid protein with 41 % similarity to theα/β hydrolase fold-3 domain protein of Burkholderia sp. The RHgene34 protein contains conserved motifs of esterases/lipases, such as HGGG (residues 95-98),GxSxG (residues 167 - 171) and the putative catalytic triad composed of Ser157,Asp255 and His285. The gene was cloned and expressed in pET21a(+), and transformed into Escherichia coli Rosetta. p-Nitrophenol (p-Np) fatty acyl esters of different carbon chain lengths were used for kinetic characterisation of RHgene34. Kinetic analysis revealed that RHgene34 had a broad range activity on the p-Npesters, from C2 - C14. RHgene34 operates optimally at 45 ºC, pH 9.0 and has a half life of 30 mins at 45 ºC. This study demonstrates that functional screening combined with the sequence analysis is a useful approach for isolating novel enzymes from ametagenome.
5

Effects of Lipolytic and Antilipolytic Agents on Glycerol and Free Fatty Acid Release from Isolated Adipocytes of Normal and Diabetic Rats

SAKAMOTO, NOBUO, KOH, NAOKI, FUKASAWA, HIDEO, KIMURA, MASAO, KAKUTA, HIRONOBU, HOTTA, NIGISHI, KAMEI, IZUMI 03 1900 (has links)
No description available.
6

Spectroscopic studies of apolipoprotein e and the low-density lipoprotein receptor /

Clayton, Daniel John. January 2001 (has links) (PDF)
Thesis (Ph. D.)--University of Queensland, 2002. / Includes bibliographical references.
7

Studium produkce lipáz pomocí vybraných mikroorganizmů / Study of lipase production using selected microorganisms

Rošková, Hana January 2014 (has links)
Diploma thesis was aimed at studying of lipolytic activity of seven microorganisms, at which this activity was previously assumed or already confirmed. Theoretical part deals with the general characteristics of lipoyltic enzymes, specifics of microbial lipases and their industrial application, with an emphasis on food industry. Experimental part deals with the study of seven microorganisms and their lipolytic activity at the presence of a carbohydrate (glucose) or a lipid (canola oil) or both, as a source of carbon. For further testing were singled out Rhodotorula minuta nad Yarrowia lipolytica. This yeasts were subsequently tested for lipolytic aktivity at the presence of different vegetable oils (olive, sunflower, canola oil and waste cooking oil), nitrogen sources of organic and inorganic origin (urea, yeast extract, amonnium chloride, amonnium sulfate) and also a addition of detergent, which is commonly used in food industry or other food facilities.
8

Prospecção de genes codificadores de enzimas lipolíticas em biblioteca metagenômica de consórcio microbiano degradador de óleo diesel. / Screening for lipolytic enzyme codification genes in a metagenomic library of consortia specialized in diesel oil degradation.

Pereira, Mariana Rangel 03 March 2011 (has links)
As enzimas lipolíticas vêm atraindo atenção no mercado global devido ao enorme potencial biotecnológico, como: na formulação de detergentes; na indústria de couro; produção de cosméticos, fármacos, aromas, biodiesel, etc. O objetivo deste trabalho foi prospectar genes codificadores de enzimas lipolíticas em biblioteca metagenômica de um consórcio microbiano degradador de óleo diesel. A seleção foi feita pela atividade lipolítica através do cultivo dos clones em placa de petri e a avaliação foi pela observação de halo ao redor da colônia, sendo positiva para 30 clones dentre os quais dois se destacaram. Estes dois clones foram selecionados e subclonados. Os DNAs das sub-bibliotecas foram sequenciados, gerando um contig completo para cada clone. Através do ORF Finder foi identificado cinco ORFs de esterase/lipase, dentre as quais uma alcançou 58% de identidade com uma bactéria não cultivável. As árvores filogenéticas indicam que duas ORFs são similares à família IV das enzimas lipolíticas, enquanto que as outras três ORFs à família V. / Lipolytic enzymes have been attracting global market attention because they show enormous biotechnological potential. The present work was done as an attempt to find genes which codify lipolytic enzymes in a metagenomic library composed of diesel oil degradation microbe consortia. Clones were selected according to lipolytic activity and were then evaluated after cultivation in Petri dishes by observation of halo formation around the colonies. 30 clones produced halo formations and were identified as positives, two of which showed prominent results. These two were then selected and sub cloned. DNA from the sub libraries was sequenced, generating a complete contig for each clone. Using the ORF Finder five esterase/lipase ORFs were identified, with one of these attaining 58% of identity to a non cultivatable bacteria species. Assessment of the cladograms showed that two ORFs were similar to lipolytic enzyme family IV, while the other three ORFs were similar to family V.
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Produção de enzimas lipolíticas por bactérias isoladas de sistemas de tratamento biológico de efluentes / Production of lipolytic enzymes by bacterials isolated from biological effluent treatment systems

Furini, Graciane January 2017 (has links)
Lipases são enzimas que hidrolisam especificamente óleos e gorduras, podendo ser de grande interesse para vários segmentos industriais como têxtil, cosméticos, alimentícios e tratamento de efluentes com elevada carga de gordura. O objetivo do presente trabalho foi avaliar a produção de complexos enzimáticos lipolíticos produzidos por microrganismos isolados de um sistema de tratamento biológico de efluentes de um hotel, visando obter lipases com potencial para emprego em processos biotecnológicos. Para a seleção do microrganismo lipolítico foram utilizados 65 isolados bacterianos oriundos do efluente bruto e tratado da wetland e da caixa de gordura do restaurante do hotel. A produção de lipase foi testada em placa em meio de cultivo acrescido de azeite de oliva e rodamina B, incubados a 25°C e 30ºC por 24h-48h e as placas observadas sob luz UV 350 nm. Deste total de isolados, sete oriundos da wetland e 22 isolados da caixa de gordura foram positivos para lipase, pois apresentaram halos fluorescentes alaranjado. Os isolados foram testados novamente em placa, porém numa condição mais estressante em termos de nutrientes para induzir a utilização do azeite e assim selecionar as melhores produtoras de enzimas Desses, 12 isolados apresentaram atividade lipolítica e aquele que apresentou halo mais evidente e maior foi selecionado para ensaios com crescimento em cultura submersa em agitador orbital e biorreator em três substratos diferentes (azeite de oliva, óleo de semente de uva e óleo de canola) para a determinação da atividade enzimática. As bactérias lipolíticas identificadas bioquimicamente foram do gênero Enterobacter, Acinetobacter, Pseudomonas, Klebsiella e Burkholderia. Os ensaios em cultura submersa em biorreator nos três substratos avaliados apresentaram a máxima produção de lipase após 12 horas de cultivo; azeite de oliva 0,358 U/mL.min-1, óleo de semente de uva 0,352 U/mL.min-1 e óleo de canola 0,348 U/mL.min-1. Em cultivo submerso em agitador orbital, o meio de cultivo na presença de Tween 80, apresentou melhor atividade enzimática que o meio de cultivo sem a presença do emulsionante. A identificação molecular do isolado apresentou identidade com o gênero Acinetobacter baylyi. A análise por SDS-PAGE sugere uma lipase com massa molecular de 35 kDa. / Lipases are enzymes that specifically hydrolyze oils and fats and may be of great interest to various industrial segments such as textiles, cosmetics, food and effluent treatment with the high fat load. The aim of this work was to evaluate the production of lipolytic enzymatic complexes produced by microorganisms isolated from a biological treatment system of effluents of a hotel, aiming to obtain lipases with potential for use in biotechnological processes. To select the best lipolytic microorganism, we used 65 bacterial isolates, recovered from the raw and treated effluent of the wetland system and from the fat tank of the hotel restaurant. Lipase production was evaluated in culture medium supplemented with olive oil and rhodamine B, incubated at 25°C and 30°C for 24h-48h. The grown colonies were observed under UV light at 350 nm. Out of that, seven isolates from wetland and 22 isolated from the fat tank were positive for lipase as they exhibited orange fluorescent halos. The isolates were again seeded on medium with a stressful nutrient condition to induce the use of olive oil and thus select the best enzyme producers Of these, 12 isolates presented lipolytic activity, and the one that showed the most evident halo was selected for assays in submerged culture in an orbital shaker and bioreactor using three different substrates (olive oil, grape seed oil, and canola oil). The lipolytic bacteria identified were of the genus Enterobacter, Acinetobacter, Pseudomonas, Klebsiella, and Burkholderia. The assays run in the bioreactor showed the maximum lipase production after 12 hours of culture with the three substrates evaluated; 0,358 U/mL.min-1 in olive oil, 0,352 U/mL.min-1 grape seed oil, and 0.348 U/mL.min-1 canola oil. The results obtained in an orbital shaker showed better enzymatic activities in the presence of Tween 80 on the culture medium. The molecular identification showed identity with the genus Acinetobacter baylyi. Analysis by SDS-page suggests a lipase with a molecular mass of 35 kDa.
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Produção de enzimas lipolíticas por bactérias isoladas de sistemas de tratamento biológico de efluentes / Production of lipolytic enzymes by bacterials isolated from biological effluent treatment systems

Furini, Graciane January 2017 (has links)
Lipases são enzimas que hidrolisam especificamente óleos e gorduras, podendo ser de grande interesse para vários segmentos industriais como têxtil, cosméticos, alimentícios e tratamento de efluentes com elevada carga de gordura. O objetivo do presente trabalho foi avaliar a produção de complexos enzimáticos lipolíticos produzidos por microrganismos isolados de um sistema de tratamento biológico de efluentes de um hotel, visando obter lipases com potencial para emprego em processos biotecnológicos. Para a seleção do microrganismo lipolítico foram utilizados 65 isolados bacterianos oriundos do efluente bruto e tratado da wetland e da caixa de gordura do restaurante do hotel. A produção de lipase foi testada em placa em meio de cultivo acrescido de azeite de oliva e rodamina B, incubados a 25°C e 30ºC por 24h-48h e as placas observadas sob luz UV 350 nm. Deste total de isolados, sete oriundos da wetland e 22 isolados da caixa de gordura foram positivos para lipase, pois apresentaram halos fluorescentes alaranjado. Os isolados foram testados novamente em placa, porém numa condição mais estressante em termos de nutrientes para induzir a utilização do azeite e assim selecionar as melhores produtoras de enzimas Desses, 12 isolados apresentaram atividade lipolítica e aquele que apresentou halo mais evidente e maior foi selecionado para ensaios com crescimento em cultura submersa em agitador orbital e biorreator em três substratos diferentes (azeite de oliva, óleo de semente de uva e óleo de canola) para a determinação da atividade enzimática. As bactérias lipolíticas identificadas bioquimicamente foram do gênero Enterobacter, Acinetobacter, Pseudomonas, Klebsiella e Burkholderia. Os ensaios em cultura submersa em biorreator nos três substratos avaliados apresentaram a máxima produção de lipase após 12 horas de cultivo; azeite de oliva 0,358 U/mL.min-1, óleo de semente de uva 0,352 U/mL.min-1 e óleo de canola 0,348 U/mL.min-1. Em cultivo submerso em agitador orbital, o meio de cultivo na presença de Tween 80, apresentou melhor atividade enzimática que o meio de cultivo sem a presença do emulsionante. A identificação molecular do isolado apresentou identidade com o gênero Acinetobacter baylyi. A análise por SDS-PAGE sugere uma lipase com massa molecular de 35 kDa. / Lipases are enzymes that specifically hydrolyze oils and fats and may be of great interest to various industrial segments such as textiles, cosmetics, food and effluent treatment with the high fat load. The aim of this work was to evaluate the production of lipolytic enzymatic complexes produced by microorganisms isolated from a biological treatment system of effluents of a hotel, aiming to obtain lipases with potential for use in biotechnological processes. To select the best lipolytic microorganism, we used 65 bacterial isolates, recovered from the raw and treated effluent of the wetland system and from the fat tank of the hotel restaurant. Lipase production was evaluated in culture medium supplemented with olive oil and rhodamine B, incubated at 25°C and 30°C for 24h-48h. The grown colonies were observed under UV light at 350 nm. Out of that, seven isolates from wetland and 22 isolated from the fat tank were positive for lipase as they exhibited orange fluorescent halos. The isolates were again seeded on medium with a stressful nutrient condition to induce the use of olive oil and thus select the best enzyme producers Of these, 12 isolates presented lipolytic activity, and the one that showed the most evident halo was selected for assays in submerged culture in an orbital shaker and bioreactor using three different substrates (olive oil, grape seed oil, and canola oil). The lipolytic bacteria identified were of the genus Enterobacter, Acinetobacter, Pseudomonas, Klebsiella, and Burkholderia. The assays run in the bioreactor showed the maximum lipase production after 12 hours of culture with the three substrates evaluated; 0,358 U/mL.min-1 in olive oil, 0,352 U/mL.min-1 grape seed oil, and 0.348 U/mL.min-1 canola oil. The results obtained in an orbital shaker showed better enzymatic activities in the presence of Tween 80 on the culture medium. The molecular identification showed identity with the genus Acinetobacter baylyi. Analysis by SDS-page suggests a lipase with a molecular mass of 35 kDa.

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