Produção de penicilinamidahidrolase por Escherichia coli : cinetica de crescimento e produção da enzima; dependencia da transferencia de oxigenio na intensidade de agitação e aeração, amplificação de escala

Orientador: Fumio YokoyaTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos e AgricolaResumo: Foi estudada a produção de penicílinainidahidrolase (PAH) por fermentação com Escherichia coli ATCC 3637, em fermentadores de laboratório, banco e piloto. No meio de fermentação empregado, baseado em água de maceração de milho, a cinética de crescimento apresenta duas fases exponenciais sucessivas. A mudança de uma a outra é provocada pela limitação de oxigênio dissolvido, A programação do incremento da taxa de transferência de oxigênio em função do tempo controla a duração relativa de ambas as fases. A velocidade específica de crescimento no primeiro período (µI) foi bem maior que no segundo (µII) - A produção de PAH segue uma cinética exponencial, ficando associada ao crescimento celular e sendo maior quanto menor o valor de µII. Quando µ tende a ymax. não se tem o acumulo de enzima. São condições necessárias para a produção de PAH a adição de ácido fenilacético como indutor e a manutenção da concentração de oxigênio dissolvida a valores muito baixos. Foi desenvolvido ura modelo cinético para a produção de PAH, baseado nas variáveis mencionadas. A fermentação foi ampliada de escala de um fermentador banco com 10 litros de meio até um fermentador piloto com 15 a 20 vezes este volume de meio de fermentação. Entre ambas unidades não se manteve similaridade geométrica. O critério seguido na ampliação da escala foi a de manter as velocidades tangenciais no agitador na mesma ordem para o modelo e o protótipo, a porcentagem da tensão de oxigênio dissolvido próxima de zero, ajustando a taxa de transferência de oxigênio máxima e o coeficiente volumétrico de transferência de massa (k1a) mediante a taxa de aeração. De uma análise dos resultados obtidos experimentalmente concluiu-se que (k1a) ou a área interfacial (a) podem ser descritos mediante as correlações de Yoshida para k1a ou de Calderbank para a, desde que seja substituído nas mesmas a velocidade de aeração associada com a secção transversal do fermentador (Vs) pela velocidade de aeração associada com a secção total dos orifícios do distribuidor. Os rendimentos de PAH foram da mesma ordem em escala banco e pilotoAbstract: Penicillinamidohiydrolase (PAH) production by Escherichia coli ATCC 9637 was studied in laboratory, bench and pilot scale fermentors. Using a culture medium based on corn steep liquor, it was shown that kinetics of growth has two succesive exponential phases. The shift from one to the other is caused by limitation of dissolved oxygen. The relative length of those phases can be controlled by programming the increment of oxygen transfer rate with time. The specific growth rate during the first period is larger than the second period. PAH production follows .exponential kinetics and it is associated with cell growth being larger if the second growth phase shows the slower rate (µII). When the value approaches the µmax no enzyme accumulation was observed. The conditions required for PAH production were the addition of phenylacetic acid as inducer and the maintenance of dissolved oxygen concentration at very low level. A kinetic model for the production of PAH was developed based on the variables above mentioned. Fermentation was scaled-up from a bench fermentor with ten liters of medium to a fermentor with 15-20 times larger volume. Geometric similarity was not maintained between those units. The criteria followed for scaling-up were to maintain a similar peripheral velocity at the tip of the impeller between model and prototype, and the percent of dissolved oxygen tension near zero. Adjustment of the maximum oxygen transfer rate and volumetric mass transfer coefficient (k1a) was acomplished by changing air rate. It was concluded, from experimental data, that (k1a)or the interfacial area (a) can be described by the correlation of Yoshida for k1a or Calderbank for a, if the surface air velocity at the cross section of the fermentor (Vs) is replaced by air velocity at the total area of holes in the sparger. Yield of PAH obtained in bench and pilot scale fermentor was the sameDoutoradoDoutor em Ciências e Tecnologia de Alimento

Process Of Extractive Fermentation And Use Thereof

PROCESSO DE FERMENTAÇÃO EXTRATIVA E USO DO MESMO. A presente invenção se refere a um processo de fermentação extrativa. Mais especificamente, a presente invenção se refere a um processo de fermentação extrativa que emprega um único solvente capaz de remover produtos e inibidores simultaneamente, além de resfriar o meio. Além disso, apresente invenção se refere ao uso do processo. Especificamente, a presente invenção se refere ao uso do biodiesel, tais como biodiesel de óleo de soja, de milho, de mamona, de dendezeiro, de macaúba, de macaúba, entre outros, como agente extrator biocompatível para a extração in-situ de produto e/ou componentes inibidores da fermentação, especificamente o produto da fermentação e compostos inibidores presentes nos mostos provenientes de processos hidrolíticos de matéria-prima lingocelulósica, amilácea, carboidratos diretamente extraíveis, melaços fortemente esgotados com alto conteúdo de inibidores como, entre outros, biotina, ou mostos similares ricos em carboidratos, como também para o resfriamento do meio da fermentação pelo próprio solvente.BRPI1105142 (A2)C12P7/06B01D3/34B01D11/04BR2011PI05142C12P7/06B01D3/34B01D11/0

Challenges and opportunities in lactic acid bioprocess design—From economic to production aspects

Lactic acid is an already consolidated bioproduct in the world market. It has many applications, such as: the production of biodegradable polymers, substitution of plastics from oil, and new uses in medicine. Besides this, new applications are being discovered every year, especially in chemical industries as a building-block molecule. The lactic acid market is in constant growth and the fact that the final products are able to comply with environmental laws as green, renewable and biodegradable products contributes to the tendency of continuous growth in the next few years. With all of this in mind, this paper will explore the main aspects of the sector, as well as market tendencies, production chain, and innovation133219239FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2013/26290-

Sistema E Processo Para Monitoramento De Processos De FermentaÇÃo

SISTEMA E PROCESSO PARA MONITORAMENTO DE PROCESSOS DE FERMENTAÇÃO. O presente pedido de patente de invenção refere-se a um processo e um sistema para monitoramento de processos de fermentação. A invenção proposta tem seu campo de aplicação no monitoramento em tempo real do processo de fermentação alcoólica, fornecendo o instante ideal para interrupção do processo fermentativo, de forma a otimizar o processo produtivo. As principais vantagens do sistema da presente invenção são decorrentes da utilização da própria fibra óptica como elemento sensor pois, estas apresentam tamanho e peso reduzidos, baixa reatividade em meios agressivos, imunidade eletromagnética a riscos em ambientes potencialmente explosivos. Além disso, para a aplicação descrita no presente relatório, a fibra óptica possui também a vantagem da facilidade de acoplamento no biorreator, a aquisição dos dados pode ser feita a distância e o sistema não requer a utilização de nenhum tipo de fibra óptica especial, sendo que as fibras ópticas comerciais, utilizadas principalmente em telecomunicações, são totalmente adequadas. O processo proposto apresenta como vantagem monitoramento em tempo real do processo de fermentação alcoólica.BR102013006864 (A2)G01N21/17C12M1/34BR20131006864G01N21/17C12M1/3

Second-generation (2g) lactic acid production and new developments – a mini-review

Lactic acid (LA) production is already a global reality. Its applications cover the most diverse industrial sectors and have rapidly consolidated in recent years. Currently, the most prominent use of LA is the production of polylactic acid to replace plastics from the petrochemical industry. A great part of this rapid change is due to the rising worldwide concerns about the excess of non-degradable plastics used daily and the accumulation of this material in nature. In this scenario, LA production becomes even more relevant when considering its production from renewable raw materials, especially second-generation (2G) substrates, such as lignocellulosic biomass. This reduces the human dependence on oil for both energy and fuel production, as well as for the production of plastics and other chemicals since LA is still one of the most relevant building block chemicals. Nowadays it is possible to produce LA from the most diverse 2G-substrates available around the world. Thus, LA production by fermentation of 2G-sugars can be associated with several existing biorefinery models, such as for biofuels and chemicals production. In this scenario, for example, it is possible to associate LA production with ethanol production in a biorefinery model, producing 1G-ethanol, 2G-LA, sugar for food, and electricity. This kind of approach may represent a break from current production model of energy and chemicals to a more sustainable and democratic scenario, including new players in the world market and reducing the dependence of other countries to supply oil and its derivates, especially when associated with new and powerful genetic engineering tools. Front this scenario, this review presents the state of the art of 2G-LA production80223228FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2019/00380-4; 2008/57873-

Polymer grade L-lactic acid production from sugarcane bagasse hemicellulosic hydrolysate using bacillus coagulans

Lactic acid has applications in many different sectors such as food, chemicals and polymers. Its production from lignocellulosic materials has been highlighted as a possibility to overcome barriers in the production chain such as decreasing process costs and reaching better environmental performance, besides adding value to biomass. However, the needed pre-treatment that lignocellulosic material goes through to release fermentable sugars also produces inhibitor compounds for the microorganisms, such as HMF and furfural. In this work, a Bacillus coagulans strain is evaluated as an L-lactic acid producer using sugarcane bagasse hemicellulosic hydrolysate. L-Lactic acid production reached 55.99 g/L, 0.87 g/g of yield, 1.7 g/ L h −1 of productivity and 99.4% of isomeric purity. Simultaneous bio-detoxification removed 100% of HMF and furfural without any treatment prior. To the best of our knowledge, it is the first time that B. coagulans is used to lactic acid production and bio-detoxification tests62631FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2013/26290-5; 2016/14830-

Detoxification of sugarcane-derived hemicellulosic hydrolysate using a lactic acid producing strain

Furfural and HMF are known for a negative impact in different bioprocesses, including lactic acid fermentation. There are already some methods described to remove these inhibitory compounds from the hydrolysates. However, these methods also reduce the yield of sugars from the hydrolysis and increase the process costs. In this work, the detoxification of sugarcane-derived hemicellulosic hydrolysate was performed by Lactobacillus plantarum during the fermentation time. At the end of the fermentation, a decrease of 98% of furfural and 86% of HMF and was observed, with a final lactic acid titer of 34.5 g/L. The simultaneous fermentation and bio-detoxification simplify the process and reduce operational costs, leading to economic competitiveness of second-generation feedstock for lactic acid production2785663FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESP2013/26290-

SACCHARIFICATION OF SUGARCANE BAGASSE FOR ETHANOL PRODUCTION USLNG THE ORGANOSOLV PROCESS

Abstract THE SCOPE of this work is the reduction of the production cost of fuel ethanol, through hydrolysis of surplus bagasse from sugar mills with annexed distilleries. Ethanol could be obtained froin the sugar liquor af3er fernsentation and distillation. An Organosolv process using an ethanol-water mixture as solvent and diluted sulfuric acid (0.1-0.25%) as the reaction catalyst was developed. Reaction was done at 170-185°C and 20-25 kg/cm2. The work was done on a bench scale unit (20 kglh of raw bagasse). Processing for conversion to hexose and pentose sugars was in a short residence time reactor. Reaction products were totally dissolved in the organosolv solvent. Solvent was removed and sugar liquor recovered after removal of precipitated lignin. Results obtained in the pilot plant were: global conversion of bagasse in total reducing sugars was 59%, being sugar content in liquor 80 g/L, after 10-12 ininutes of residence time. The final liquor containing reducing sugars was fermented successfully to ethanol after blending with cane juice and molasses. Fernsei~tation used a yeast strain adapted to ferment the hexose fraction despite the high concentration of inhibitors present, such as short chain organic acid, furfural and hydroximetilfurfural. The ratio of hydrolysis liquor to cane sugar components was adjusted to match a maximum level of inhibitor in must. Results of the bench unit were scaled up to a 5000 litrestday pilot and demonstration unit (PDU). Our preliminary feasibility calculations allow us to forecast that the ethanol produced by the organosolv process should be competitive with current ethanol produced directly from cane juice and molasses. There is also a large potential for cost reduction once the hydrolysis reaction is optimised. Developme~st of fermentation of the pentose sugar fraction to ethanol, to be done in the future, will improve the economy of this process. The organosolv technology for bagasse hydrolysis is of medium complexity and Brazil or other developing countries can rapidly have access to it. This process could increase the production of alcohol in Brazil, using the same cane planted area and reducing the production costs, besides allowing the use of many other lignocellulose residues from agriculture, forest industry and others for ethanol production