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Characterization of The Viable but Non-Culturable Legionella pneumophila in Water and the Role of 3-Hydroxybutyrate Dehydrogenase in Its FormationAl-Bana, Badii 16 September 2013 (has links)
Legionella pneumophila, the causative agent of Legionnaires’ disease (LD), is an intracellular pathogen of freshwater protozoa that can also persist in the environment as a free-living bacterium. L. pneumophila has many morphological forms that fit within a developmental cycle. In water, L. pneumophila enters into a viable but non-culturable (VBNC) state that is largely uncharacterized. VBNC cells were produced from two developmental L. pneumophila forms, stationary phase forms (SPFs) and mature infectious forms (MIFs) by suspension in double deionized (dd) or tap-water at 45°C. Electron microscopy results showed that VBNC cells have a unique morphology and that in tap water they lose their poly 3-hydroxybutyrate inclusion bodies. Both SPFs and MIFs lost culturability faster in dd- than in tap water, and addition of salts to dd-water prolonged L. pneumophila culturability and enhanced viability. However, MIFs retained higher viability in dd- and tap water (85% and 51%, respectively) than SPFs (5% and 20%, respectively) as determined by the BacLight vital stain. Only ~1 VBNC cell out of 105 of those produced from SPFs in tap water regained culturability via infection of Acanthamoeba. All VBNC cells, except for those produced from SPFs in dd-water, resisted both digestion inside Tetrahymena spp. and detergent-mediated lysis. SDS-PAGE analysis and shotgun proteomics revealed a number of VBNC cell specific proteins; one of these was 3-hydroxybutyrate dehydrogenase (BdhA), which is involved in the metabolism of poly 3-hydroxybutyrate inclusion bodies. A bdhA mutant showed an early loss of culturability and a dramatic decrease in viability as compared to the parent strain, and complementing the mutant with a functional bdhA gene restored the parent's strain phenotypes. In conclusion, VBNC L. pneumophila has a distinct morphology and physiology that varies according to the developmental stage and the environmental conditions used to produce such VBNC cells. VBNC cells have a different protein profile and morphology than the culturable cells, suggesting that this state constitutes a distinct differentiated form within the developmental cycle of L. pneumophila. BdhA seems to influence L. pneumophila survival and hence VBNC cell formation. Collectively, the results from this study provide a better understanding of L. pneumophila VBNC form and the factors influencing its formation.
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Polyhydroxyalkanoates production by a bacterium isolated from mangrove soil samples collected from Quang Ninh province / Vi khuẩn sinh tổng hợp polyhydroxyalkanoates phân lập từ đất rừng ngập mặn tỉnh Quảng NinhDoan, Thuoc Van, Nguyen, Binh Thi 15 July 2013 (has links) (PDF)
A PHA producing bacterium (strain QN271) was selected from mangrove soil samples collected from Quang Ninh province by using the Nile red dying technique. PHA accumulation in the selected bacterium strain was confirmed by transmission electron microscope. With the exception of maltose or sucrose, the bacterium strain was found to be able to synthesize PHA from various carbon sources (glucose, xylose, fructose, glycerol, and glucose plus propionate). The strain accumulated poly(3-hydroxybutyrate) from glucose, fructose, xylose, and glycerol whereas poly(3-hydroxybutyrate-co-3-hydroxyvalarate) was produced when a combination of glucose and propionate was included in the culture medium. Fructose was found to be most suitable substrate for PHA synthesis by strain QN271. PHA content of 63.3% and CDW of 6 g/L were obtained after 32 hrs of cultivation in fructose medium. / Chủng vi khuẩn có khả năng sinh tổng hợp PHA đã được phân lập từ đất rừng ngập mặn tỉnh Quảng Ninh nhờ kỹ thuật nhuộm với Nile red. Ảnh quan sát dưới kính hiển vi điện tử dẫn truyền chứng tỏ rằng chủng vi khuẩn này có khả năng tích lũy lượng lớn PHA trong tế bào. Chủng vi khuẩn tuyển chọn có khả năng sinh tổng hợp PHA từ nhiều nguồn các bon khác nhau như glucose, xylose, fructose, glucerol, glucose và propionate nhưng không có khả năng tổng hợp PHA từ maltose hoặc saccharose. Chủng vi khuẩn tuyển chọn tổng hợp poly (3-hydroxybutyrate) từ các nguồn các-bon như glucose, xylose, fructose, hay glycerol, trong khi đó poly (3-hydroxybutyrateco- 3-hydroxyvalarate) sẽ được tổng hợp khi phối hợp sử dụng hai nguồn các-bon (glucose và propionate). Fructose là nguồn các-bon tốt nhất cho chủng QN271 sinh tổng hợp PHA, khi nuôi cấy trong môi trường có fructose chủng vi khuẩn này có thể tạo ra lượng sinh khối là 6 g/L trong đó có chứa 63.3% PHA sau 32 giờ.
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Blendas com poli(3-hidroxibutirato) (PHB) e copolimeros aleatorios = comportamento de fases e cinetica de cristalização / Blends of poly(3-hydroxybutyrate) and random copolymers : phase behavior and crystallization kineticsTaba, Eduardo dos Santos 15 August 2018 (has links)
Orientador: Maria Isabel Felisberti / Tese (doutorado) - Universidade Estadual de Campinas, Instituto de Quimica / Made available in DSpace on 2018-08-15T06:00:08Z (GMT). No. of bitstreams: 1
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Previous issue date: 2009 / Resumo: Blendas do polímero biodegradável poli(hidroxibutirato) foram preparadas com os seguintes copolímeros aleatórios: poli(estireno-co-acrilonitrila)-SAN; poli(estireno-co-metacrilato de 2-hidróxietila)-S-Hema; poli(metacrilato de metila-co-vinil fenol-P(MMA-co-VPh). Os copolímeros SAN foram sintetizados via copolimerização em massa, enquanto os copolímeros S-Hema foram sintetizados pela copolimerização em solução utilizando DMF como solvente. Os copolímeros foram caracterizados por ressonância magnética nuclear (RMN) de H e C, cromatografia de permeação em gel (GPC), calorimetria exploratória diferencial (DSC) e análise dinâmico-mecânica (DMA). As blendas foram preparadas dissolvendo-se os polímeros em um bom solvente e adicionando-se à solução resultante um mau solvente para a coagulação dessas blendas. As análises das blendas por DSC, DMA e microscopia eletrônica de varredura (SEM) mostraram que todas elas são imiscíveis. A cinética de cristalização do PHB em blendas P(MMA-co-VPh)/PHB e SAN/PHB foi estudada por DSC. Esses estudos mostraram que a presença do copolímero P(MMA-co-VPh) causa a diminuição da taxa de cristalização do PHB e aumenta a energia de ativação do processo de cristalização do PHB. Para as blendas PHB/SAN, o efeito do copolímero em diminuir a taxa de cristalização do PHB é menor que nas blendas PHB/P(MMAco- VPh). Além disso, o teor de acrilonitrila no copolímero pouco afeta a taxa de cristalização do PHB. Os expoentes de Avrami ¿n¿ determinados para a cristalização do PHB nas blendas P(MMA-co-VPh)/PHB e SAN/PHB são aproximadamente iguais ao expoente ¿n¿ para o PHB puro indicando que o mecanismo de cristalização do PHB não se altera nas blendas. Sendo assim, este trabalho possibilitou o entendimento de aspectos importantes referentes ao comportamento de fases e à cinética de cristalização de blendas contendo PHB associado a copolímeros aleatórios.Os estudos cinéticos podem colaborar para a compreensão do comportamento de cristalização de copolímeros/PHB em equipamentos de processamento. Além disso, esse estudo pode ajudar na escolha do copolímero e de sua composição para um melhor controle da cristalização do PHB nessas blendas / Abstract: Blends of biodegradable poly(hydroxybutyrate) were prepared with the following random copolymers: poly(styrene-co-acrylonitrile)-SAN; poly(styrene-co-2-hydroxyethylmethacrilate)-S-Hema; poly(methylmethacrylateco- vinylphenol)-P(MMA-co-VPh).SAN copolymers were synthesized by bulk copolymerization; while S-Hema copolymers were synthesized by solution copolymerization using DMF as solvent. The resulting copolymers were characterized by nuclear magnetic resonance (RMN) of H and C, gel permeation chromatography (GPC), differential scanning calorimetry (DSC) and dynamic-mechanical analysis (DMA). Blends were prepared using coprecipitation method where a binary homogeneous soilution was added to a large volume of non solvent. DSC, DMA and scanning electron microscopy (SEM) analysis revealed that all the blends are immiscible in the entire composition range. Despite the immiscible blends, DSC analysis show that the copolymers interferes in the PHB crystallization, being capable of suppress this process in some blends. Crystallization kinetics of the PHB in PHB/P(MMA-co-VPh) and PHB/SAN blends was studied by DSC and the results revealed that the presence of P(MMA-co-VPh) copolymer decreases the PHB crystallization rate and increases the activation energy for the overall crystallization process. In respect to PHB/SAN blends, the influence of the copolymer in decrease the PHB crystallization rate is lower than the observed in PHB/P(MMA-co-VPh) blends. Moreover, the acrylonitrile concentration in the copolymer causes little effect in the PHB crystallization rate. The Avrami exponent ¿n¿ determined for the crystallization process in PHB/P(MMA-co-VPh) and SAN/PHB blends are approximately equal to the ¿n¿ exponent for pure PHB indicating that the crystallization mechanisms for pure PHB do not change in the blends. This work made possible the understanding of important aspects of the phase behavior and crystallization kinetics of random copolymers/PHB blends. The crystallization kinetics studies can collaborate for the understanding of the behavior of crystallization of these blends in processing equipments. Moreover, this study can help in the choice of the copolymer and its composition for better control of the crystallization of PHB in these blends / Doutorado / Físico-Química / Doutor em Ciências
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Příprava a charakterizace biologicky rozložitelných kompozitů na bázi polyhydroxyalkanoátů a kávové sedliny / Preparation and characterization of biodegradable composites based on polyhydroxyalkanoates and coffee groundsCisáriková, Barbora January 2020 (has links)
This diploma thesis was focused on the preparation and characterization of biodegradable composites based on poly-3-hydroxybutyrate (PHB) and spent coffee grounds (SCG). The aim was to prepare biocomposites based on PHB and coffee grounds. Biocomposites were prepared by solution casting in chloroform. Films with a thickness of about 150-200 µm were prepared with the addition of 20% unmodified and modified coffee grounds. The PHB/SCG films were modified by an addition of a plasticizer and a crosslinking agent or by blending with polybutylene adipate terephthalate (PBAT). The thermal properties of the films were determined by differential scanning calorimetry. The compatibility filler/polymer matrix compatibility was assessed morphologically with the naked eye. It has been found that the direct addition of spent coffee grounds to the PHB is not suitable due to the deterioration of the morphology of the film surfaces and the formation of aggregates. The compatibility between the spent coffee grounds and the PHB was improved after the addition of the plasticizer and the crosslinking agent. However, the highest compatibility between SCG and PHB was achieved in the case of esterified coffee grounds. Another way to improve the resulting morphological properties of the films thus prepared was to use coffee grounds fractions.
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Studium reologických vlastností a biodegradace poly (3-hydroxybutyrátu-co-4-hydroxybutyrátu) / Rheological properties and biodegradation of Poly (3-hydroxybutyrate-co-4-hydroxybutyrate)Černeková, Nicole January 2021 (has links)
This diploma thesis deals with the characterization of rheological properties and biodegradation of a poly(3-hydroxy-co-4-hydroxybutyrate), a copolymer produced by a strain of Cupriavidus malaysiensis. The theoretical part focuses on the rheology and degradation of polymers in general. It also contains the basic characteristics of polyhydroxyalkanoates and deals in more details with the description of the studied copolymer, its properties, synthesis, decomposition and possibilities of its applications. In the experimental part, the given copolymer was investigated in its native form and also in the form of copolymer mixtures filaments, which were enriched with additives (stabilizers and plasticizers). In terms of rheology, the samples showed pseudoplastic behaviour, which was slightly limited by the plasticization of the material. The highest thermal stability observed as a function of changes in complex viscosity over time was observed in a sample of a copolymer mixture containing stabilizers and plasticizer Citrofol BII. The plasticization also caused significant changes in thermal properties, especially crystallinity, which decreased compared to the native copolymer. In vitro degradation studies of samples in the form of films prepared by dissolving copolymer mixtures in chloroform were performed in phosphate buffer with lipase, in simulated body fluid and in synthetic gastric juice. The course of the process itself was characterized by the method of determining the molecular weight (SEC-MALLS) and the weight loss over time (gravimetrically). The results showed that the analysed samples are subject to degradation in all used environments. The most considerable molecular weight loss after 105 days (76 %) was observed in the sample containing stabilizers and plasticizer Citrofol BII in the environment of synthetic gastric juice. The most extensive gravimetric weight loss was attributed to the sample with stabilizers and without plasticizers, in the environment of phosphate buffer with lipase, specifically by 79%.
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Réactions Polyoléfines/ Poly (3-hydroxybutyrate-co-hydroxyvalérate) : des mélanges compatibilisés aux copolymères greffés / Polyolefins/ Poly (3-hydroxybutyrate-co-hydroxyvalerate) reactions : from compatibilized blends to grafted copolymersSadik, Tarik 16 December 2011 (has links)
L’objectif de cette thèse est le développement des matériaux polymères injectables innovants issus de ressources renouvelables et répondant aux exigences techniques automobiles. Pour cela, des mélanges à base de polyoléfines (POs) et du poly(3- hydroxybutyrate-co-hydroxyvalérate) (PHBV) ont été étudiés. Ces mélanges polymères étant non-miscibles, l’optimisation de leurs propriétés par compatibilisation a été réalisée selon différentes approches. Dans un premier temps, la synthèse de polypropylène fonctionnel original avec des taux de greffage relativement élevés et sans perte de masses molaires a permis, par réaction in-situ, la compatibilisation des mélanges PP/PHBV. Dans une deuxième partie, le poly(éthylène-co- alcool vinylique) greffé-poly(3- hydroxybutyrate-co- hydroxyvalérate) a été synthétisé par extrusion réactive, ce copolymère a été utilisé ensuite comme compatibilisant pour les mélanges PE/PHBV. L’ajout de ces compatibilisants a permis d’obtenir des matériaux ayant des propriétés optimisées et une morphologie nettement plus fine que les systèmes non compatibilisés démontrant ainsi leur efficacité / The main objective of this work is the development of innovative materials from renewable resources for the automotive industry. Polymer blends of polyolefins (POs) and poly(3-hydroxybutyrate-co-hydroxyvalerate) (PHBV) were prepared through twin screw extruder process and studied. These polymer blends are non-miscible, then in order to optimize their final properties, compatibilizing agents were synthesized by reactive extrusion. On the one hand, functionalized polypropylene with various polar monomers and without significant molecular weight loss was synthesized with the aim to compatibilize PP/PHBV blends, on the other hand, poly(ethylene-co- vinylalcohol)-graft- poly(3-hydroxybutyrate-co-hydroxyvalerate) was obtained by exchange reactions in the presence of an efficient catalyst and those in order to compatibilize PE/PHBV blends. Efficiency of these compatibilizers was investigated in the last part of this study which is dedicated to POs/PHBV polymer blends compatibilization. Materials with improved properties and finer morphologies than the non- compatibilized systems were obtained thanks to the compatibilizers
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Composition and property study of adhesives based on poly(3-hydroxybutyrate-co-3-hydroxyvalerate) / Sammansättnings- och egenskapsundersökning för bindemedel baserade på poly(3-hydroxybutyrat-co-3-hydroxyvalerat)Johnsson, Nathalie January 2021 (has links)
Lim klassificeras som ett ämne som kan hålla ihop två ytor så att de motstår separation. Dagens lim kan anpassas efter vilken applikation de ska användas till och kan ha ett stort antal olika egenskaper. De fysikaliska och kemiska egenskaperna av limmet är de viktigaste faktorerna för att bilda en bra limbindning. I stort sett alla syntetiska lim består idag av polymerer, varav de flesta är petroleumbaserade. För att skapa ett mer miljövänligt alternativ undersökte denna studie tillämpningen av poly(3-hydroxibutyrat- co-3-hydroxivalerat), PHBV med 36 eller 56 vikt% HV, som huvudkomponent i ett lim. Huvudfokus ligger på hur väl PHBV är lämpligt för användning som lim och hur olika tillsatser kan förbättra dess egenskaperna. Flera olika limblandningar innehållande PHBV, mjukgörare (sebacinsyra, dimetylsebacat, etylbutanoat eller tributylcitrat) och förtjockningsmedel (Abalyn eller Foralyn) skapades och undersöktes. Ett single lap skjuvtest utfördes med kopieringspapper, filterpapper och träpinnar som vidhäftningsmaterialet, medan ett avskalningstest undersökte användningen av kopieringspappersetiketter och plastetiketter på en glasflaska samt frukt. Båda testen visar att kopieringspapper har de mest lovande egenskaperna som en adherent för användningen av ett PHBV-baserat lim, både för PHBV innehållande 36 vikts% och 56 vikts% HV. Denna slutsats kunde dras då kopieringspapperet påvisade den starkaste bindningen med limmet. Ren PHBV uppvisade lovande häftstyrka och vidhäftning som ett smältlim. Tillsatsen av sebacinsyra tillsammans med Abalyn eller dimetylsebacat med Foralyn ökade limmets häftstyrka ytterligare. Vi fann också att lim som skapats med PHBV med 36 vikt% HV ger bättre hållfasthetsegenskaper. Framtida arbete innefattar mer exakta mätmetoder för att bestämma egenskaperna hos limblandningarna. / An adhesive is classified as a substance that holds two surfaces together and resists separation. Today’s adhesives can be modified according to various application demands, obtaining a large variety of properties. The most important factors of forming a good adhesive bond are the physical and chemical properties. Essentially all synthetic adhesives consist of polymers, most of them being petroleum-based. To obtain a more environmentally friendly option, this study investigated the use of poly(3-hydroxybutyrate-co-3-hydroxyvalerate), PHBV with 36 or 56 wt% HV, as the main component in an adhesive. The main focus was to investigate on how well PHBV is suited for use as an adhesive and how different additives can improve the adhesive properties. Several different adhesive formulations containing PHBV, plasticizers (sebacic acid, dimethyl sebacate, ethyl butyrate, or tributyl citrate) and tackifiers (Abalyn or Foralyn) were created and investigated using various tests, such as single lap shear test, peel test, tackiness determination, optical analysis, and application testing. Single lap shear tests were performed using printing paper, filter paper, and wooden sticks as adherents, while peel tests explored the use of printing paper labels and plastic labels on a glass bottle and on fruit. It was determined that two PHBV adhesives, containing 36 wt% and 56 wt% of HV, performed best using printing paper as adherent. This conclusion could be drawn based on the good interaction between the adherent and the adhesive, thereby creating a strong bond. Pure PHBV with 36 or 56 wt% showed promising strength and tackiness properties as a hot-melt adhesive. The addition of sebacic acid together with Abalyn or dimethyl sebacate with Foralyn further increased the adhesive’s strength. It was also found that adhesive formulations created using a PHBV with a lower amount of HV (around 36 wt% of HV) yields better strength properties when used as an adhesive for paper labels. Future work involves more precise measurement methods to determine the properties of the adhesive formulations.
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Estudo da blenda poli(3-hidroxibutirato) / poli(etileno glicol). / Poly(3-hydroxybutyrate) / poly(ethylene oxide) blend study.Blazek, Gustavo Russo 16 December 2011 (has links)
Esse projeto visa a melhoria das propriedades mecânicas e de processabilidade do Poli(3-hidroxibutirato) (PHB) para futuras aplicações. Foram preparados filmes de blendas de PHB por dissolução em clorofórmio. Poli(etileno glicol), com massa molar de 300 (PEG) ou 4.000.000 g mol-1 (PEO), foi misturado em proporções de 5 a 30 % em massa e análises termogravimétricas (TG), de calorimetria diferencial exploratória (DSC) e ensaios dinâmico-mecânicos (DMTA) foram realizadas para avaliar a miscibilidade da blenda. Foram também analisados filmes recém-preparados e filmes envelhecidos, para avaliar o avanço da cristalização e consequente alteração das propriedades dos filmes com o passar do tempo. Para isso foram feitas comparações visuais de imagens microscópicas da morfologia das blendas, obtidas através de microscopia eletrônica de varredura (MEV). As blendas foram analisadas quanto ao grau de cristalinidade através de análise calorimétrica (DSC) e de difratometria de raio-X. As curvas de DSC mostram uma redução da Tm do PHB com a adição de PEG e PEO, o que indica uma forte interação entre os polímeros. As análises de TG mostram uma etapa principal de degradação, evidenciando a miscibilidade entre os polímeros. A TG também mostra uma sensível redução na temperatura de degradação do PHB para concentrações acima de 10 % em PEG, o que é indesejável. Através do MEV nota-se que existe também uma segregação de fases que aumenta com o tempo de estocagem do material, levando à recristalização do PHB e sua consequente fragilização. A difração de raios-X nos mostra que a adição de PEG e PEO ao PHB traz uma redução considerável para a cristalinidade do sistema, e que o aumento de 5 % para 30 % no teor de PEG é responsável por apenas uma pequena redução na cristalinidade, mas uma considerável redução na recristalização sofrida pela blenda com o tempo de estocagem. As análises de DMTA mostram que as blendas possuem módulo de armazenamento similar ao de polímeros flexíveis, evidenciando uma efetiva tenacificação do PHB. Amostras utilizando PEO apresentam maior rigidez do que as amostras contendo PEG. / This project aims the enhancement of both processability and mechanical properties of poly(3-hydroxybutyrate) (PHB) for future applications. Films of blends of PHB and poly(ethylene oxide) were prepared by chloroform solution casting and evaporation. Poly(ethylene oxide) having molar mass of 4,000,000 (PEO) or 300 g.mol-1 (PEG) was blended in proportion of 5 to 30 wt %, Thermogravimetric Analysis (TG), Differential Scanning Calorimetry (DSC) and Dynamic-Mechanic Analysis (DMTA) were performed in order to evaluate the miscibility of the blends. As-cast and aged films were also compared in order to analyze the crystallization progress and the consequent changes in the blends properties due to the aging process. For this, visual comparison was drawn between microscopy pictures obtained through scanning electron microscopy (SEM). The blends have their crystallinity degree determined by analyzing their X-ray diffraction curves. The DSC curves show a reduction at the PHBs Tm as the mass percentage of PEG increases, indicating a strong interaction in between the polymers. The TG and DTG analyses reveal a single main degradation step, what denotes miscibility in between the polymers. The TG also shows a considerable reduction of the PHBs degrading temperature for PEG concentrations over 10 %, which is undesirable. Through SEM one can note a phase segregation that increases with storaging time, leading to further crystallization of PHB and its subsequent enbrittlement. The X-ray diffraction curves show that the PEG and PEO bring a considerable crystallinity reduction to the system, and that the increasing of PEG content from 5 to 30% has only a minor effect per se, though also a considerable reduction of the perfection undergone by the system with storage time. The DMTA shows that the blends have a storage modulus similar to the one of flexible polimers, hence showing an effective PHB toughening. Samples containing PEO are more rigid than those containing PEG.
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Engineering Cyanobacteria to Convert Carbon Dioxide to Building Blocks for Renewable PlasticsJanuary 2014 (has links)
abstract: The production of monomer compounds for synthesizing plastics has to date been largely restricted to the petroleum-based chemical industry and sugar-based microbial fermentation, limiting its sustainability and economic feasibility. Cyanobacteria have, however, become attractive microbial factories to produce renewable fuels and chemicals directly from sunlight and CO2. To explore the feasibility of photosynthetic production of (S)- and (R)-3-hydroxybutyrate (3HB), building-block monomers for synthesizing the biodegradable plastics polyhydroxyalkanoates and precursors to fine chemicals, synthetic metabolic pathways have been constructed, characterized and optimized in the cyanobacterium Synechocystis sp. PCC 6803 (hereafter Synechocystis 6803). Both types of 3HB molecules were produced and readily secreted from Synechocystis cells without over-expression of transporters. Additional inactivation of the competing PHB biosynthesis pathway further promoted the 3HB production. Analysis of the intracellular acetyl-CoA and anion concentrations in the culture media indicated that the phosphate consumption during the photoautotrophic growth and the concomitant elevated acetyl-CoA pool acted as a key driving force for 3HB biosynthesis in Synechocystis. Fine-tuning of the gene expression levels via strategies, including tuning gene copy numbers, promoter engineering and ribosome binding site optimization, proved critical to mitigating metabolic bottlenecks and thus improving the 3HB production. One of the engineered Synechocystis strains, namely R168, was able to produce (R)-3HB to a cumulative titer of ~1600 mg/L, with a peak daily productivity of ~200 mg/L, using light and CO2 as the sole energy and carbon sources, respectively. Additionally, in order to establish a high-efficiency transformation protocol in cyanobacterium Synechocystis 6803, methyltransferase-encoding genes were cloned and expressed to pre-methylate the exogenous DNA before Synechocystis transformation. Eventually, the transformation efficiency was increased by two orders of magnitude in Synechocystis. This research has demonstrated the use of cyanobacteria as cell factories to produce 3HB directly from light and CO2, and developed new synthetic biology tools for cyanobacteria. / Dissertation/Thesis / Ph.D. Biological Design 2014
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Avaliação do efeito da desinfecção por agentes químicos e da irradiação gama no poli(3-hidroxibutirato)SOUZA, Anailda Maria Pereira Lopes de 14 March 2016 (has links)
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Previous issue date: 2016-03-14 / PRH-ANP/MCT / Muitos trabalhos vem norteando a utilização do poli(3-hidroxibutirato)-PHB, polímero da
família dos Poli(hidroxialcanoatos), que possui como principais características,
biodegradabilidade e biocompatibilidade, o que lhe permite aplicações na área médicohospitalar,
médico-farmacêutica, embalagens alimentícias, entre outras. O estudo sobre a
degradação de polímeros quando exposto a qualquer tipo de fonte degradativa é tema de
grande interesse científico e industrial, em razão das alterações químicas que podem ocorrer
na cadeia, podendo ocasionar a cisão (ou quebra) de ligações na cadeia principal ou em
grupos laterais, reticulação, eliminação ou substituição de cadeias laterais, reações
intramoleculares, auto-oxidação e despolimerização. A avaliação das propriedades do PHB
após um processo de esterilização, ou mesmo da estabilidade desse material quando exposto a
alguma substância química e/ou radiação gama é um importante tema de estudo pois é
necessário adequar os materiais às normas, principalmente em aplicações como embalagem
para uso médico hospitalar. Esse trabalho teve como objetivo avaliar a estabilidade à radiação
gama do PHB, bem como investigar a estabilidade desse polímero quando imerso nas
soluções de glutaraldeído 2% e etanol 70%. Os filmes de PHB foram preparados pela técnica
de solution casting. Analisou-se amostras de filmes de PHB puro; PHB irradiados a 25 kGy;
PHB imerso em glutaraldeído 2% irradiado 25 kGy e não irradiados com tempo de imersão na
solução por 2, 4, 6 e 8 horas; PHB imerso em etanol 70% (EtOH) irradiado 25 kGy e não
irradiados, com tempo de imersão na solução por 2, 4, 6 e 8 horas. As propriedades destes
foram avaliadas a partir de diversas técnicas, tais como: infravermelho com transformada de
Fourier (FTIR), ensaio de tração, calorimetria exploratória diferencial (DSC),
termogravimetria (TGA). A partir das análises por componentes principais realizadas através
dos espectros de infravermelho das amostras dos filmes de PHB puro, PHB imersos em
glutaraldeído 2%, etanol 70% e, após exposição à radiação gama, apresentaram alterações
químicas na sua estrutura, após exposição à radiação gama ou imersão nos agentes químicos
estudados. Verificou-se, portanto, que o PHB é sensível à radiação gama, apresentando como
efeito fundamental perdas de suas propriedades mecânicas, o que pode ser resultante do
processo de cisão molecular. Contudo, em relação a esterilização do polímero com
glutaraldeído e etanol, não se evidenciou perdas nas propriedades mecânicas das amostras
imersas em ambas as soluções. Quanto às propriedades térmicas, amostras de PHB imersas
em glutaraldeído 2% e etanol 70% apresentaram comportamento semelhante ao PHB puro. A
degradação térmica dos filmes de PHB ocorreu em um único estágio e em uma faixa de
temperatura que variou para a amostra PHB puro de 225,4 ºC a 295,1 ºC. O comportamento
térmico das amostras de PHB imersas em glutaraldeído e etanol observado pelas curvas de
TGA, antes e após processos de esterilização por radiação gama, foi semelhante ao
comportamento térmico do filme de PHB puro. / A lot of study have conducted the use of poly (3-hydroxybutyrate)-PHB, a polymer from the
poly (hydroxyalkanoates) family, which has main characteristics such as biodegradability and
biocompatibility that allows applications in the medical-hospital, medical-pharmaceutical and
food packaging areas. A study on polymer degradation when exposed to any type of
degradative source is a subject of great scientific and industrial interest due to the chemical
changes that may occur in the chain such as scission, crosslinking, elimination or substitution
of side chains, intramolecular reactions, auto-oxidation and depolymerization. Sterilized PHB
or even its stability when exposed to some chemical substance or gamma radiation, becomes
important topics of study to fit the standards, especially in applications such as packaging for
hospital-medical use. The objective of this work was to evaluate the PHB gamma radiation
stability and to investigate the stability of this polymer when immersed in 2% glutaraldehyde
and 70% ethanol solutions. PHB films were prepared by the technique of solution casting.
The analyzed samples were: films of pure PHB; PHB irradiated at 25 kGy; PHB immersed in
2% glutaraldehyde irradiated 25 kGy and non-irradiated, with immersion time in the solution
for 2, 4, 6 and 8 hours; PHB immersed in ethanol 70% (EtOH) irradiated 25 kGy and nonirradiated,
with immersion time in the solution for 2, 4, 6 and 8 hours. Its properties were
evaluated from several techniques such as: Fourier transform infrared (FTIR), tensile test,
differential scanning calorimetry (DSC), thermogravimetry (TGA). Based on the analysis of
the main components of different types of PHB samples (PHB immersed in 2%
glutaraldehyde, 70% ethanol and after exposure to gamma radiation) showed chemical
changes in structure, after exposure to gamma radiation or immersion in the chemical agents
studied. It was verified that the PHB is sensitive to gamma radiation, presenting as
fundamental effect losses of its mechanical properties, which may be the result of the
molecular scission process. However, in relation to the sterilization of the polymer with
glutaraldehyde and ethanol, no losses were observed in the mechanical properties of the
samples immersed in both solutions. Regarding to the thermal properties PHB samples
immersed in 2% glutaraldehyde and 70% ethanol presented similar behavior in comparison to
pure PHB. The thermal degradation of PHB films occurred in a single stage and in a
temperature range, which varied for the pure PHB sample from 225.4 ° C to 295.1 ° C. The
thermal behavior of PHB samples immersed in glutaraldehyde and ethanol prior to and after
gamma ray sterilization procedures observed by the TGA curves were similar to the thermal
behavior of the pure PHB film.
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