Produção de lipases por fungos termofílicos e mesofílicos e uso na produção de biodiesel etílico
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Data
2012-03-23
Autores
Borges, Janaina Pires [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
O biodiesel é uma alternativa ao diesel convencional que pode ser produzido a partir da reação de transesterificação entre triglicerídeos presentes em óleos vegetais ou gordura animal e alcoóis, como o metanol ou etanol. Uma rota promissora desta reação é a catálise enzimática, utilizando lipases. Este trabalho teve objetivo de selecionar cepas fúngicas com capacidade de produzir enzimas lipolíticas com propriedade de transesterificação e a imobilização destas enzimas para produção de biodiesel etílico. Para selecionar as linhagens com atividade lipolítica, foram realizados zimogramas com corante Rodamina B. Foram selecionadas três cepas fúngicas termofílicas e dezoito mesofílicas. As atividades hidrolíticas das linhagens fúngicas selecionadas foram comfirmadas com substrato cromogênico, p-nitrofenil palmitato (pNPP). As linhagens pré-selecionadas foram submetidas ao teste de transesterificação com óleo de soja e etanol, e os produtos desta reação foram analisados por GC-FID. Uma cepa fúngica identificada como Acremonium sp P24 foi selecionada para dar continuidade ao trabalho. A lipase bruta produzida a partir do bagaço de cana de açúcar foi concentrada por ultrafiltração e apresentou atividade hidrolítica ótima a 35ºC. O processo de imobilização mais eficiente para a realização de síntese foi aquele em que se utilizou as hifas do fungo Acremonium sp P24 crescidas no próprio farelo de trigo como suporte. A análise de variância para os experimentos mostrou que entre as variáveis estudadas (temperatura de reação, razão molar óleo/álcool, concentração enzimática e razão v/v óleo/hexano), somente a concentração enzimática como variável significativa e indicou a concentração de 10 % de hifas imobilizadas...
Biodiesel is an alternative to conventional diesel, which can be produced from the transesterification reaction of triglycerides found in vegetable oils or animal fat and alcohols such as methanol or ethanol. A promising route to this reaction is the enzymatic catalysis using lipases. This work aimed at selecting fungal strains capable of producing lipolytic enzymes with property of transesterification and immobilization of such enzymes for the production of ethylic biodiesel. In order to select strains with lipolytic activity, zymograms using dye Rhodamine B were performed. Three thermophilic and eighteen mesophilic fungal strains were selected. The hydrolytic activities of the selected fungal strains were confirmed by using chromogenic substrate p-nitrophenyl palmitate (pNPP). The pre-selected strains were then submitted to transesterification test with ethanol and soybean oil, and the products of such a reaction were analyzed by GC-FID. Only the fungal strain identified as Acremonium sp P24 was selected to carry out the work. The crude lipase produced from sugar cane bagasse was concentrated by ultrafiltration and showed optimal hydrolytic activity at 35°C. The most efficient immobilization process in order to carry out synthesis was the one in which hyphae of the fungus Acremonium sp P24 was used, they grown on wheat bran as support. Analysis of variance for the experiments showed that among the studied variables (reaction temperature, molar ratio of oil/alcohol, enzyme concentration and ratio v/v oil/hexane), only the enzymatic concentration as a significant variable and indicated a concentration of 10% of immobilized hyphae as an optimum point. However, experimentally speaking, the best biodiesel yield was 15.95%, using 7.75% immobilized hyphae, at 31.25°C, with 1:7.5 oil/alcohol molar ratio... (Complete abstract click electronic access below)
Biodiesel is an alternative to conventional diesel, which can be produced from the transesterification reaction of triglycerides found in vegetable oils or animal fat and alcohols such as methanol or ethanol. A promising route to this reaction is the enzymatic catalysis using lipases. This work aimed at selecting fungal strains capable of producing lipolytic enzymes with property of transesterification and immobilization of such enzymes for the production of ethylic biodiesel. In order to select strains with lipolytic activity, zymograms using dye Rhodamine B were performed. Three thermophilic and eighteen mesophilic fungal strains were selected. The hydrolytic activities of the selected fungal strains were confirmed by using chromogenic substrate p-nitrophenyl palmitate (pNPP). The pre-selected strains were then submitted to transesterification test with ethanol and soybean oil, and the products of such a reaction were analyzed by GC-FID. Only the fungal strain identified as Acremonium sp P24 was selected to carry out the work. The crude lipase produced from sugar cane bagasse was concentrated by ultrafiltration and showed optimal hydrolytic activity at 35°C. The most efficient immobilization process in order to carry out synthesis was the one in which hyphae of the fungus Acremonium sp P24 was used, they grown on wheat bran as support. Analysis of variance for the experiments showed that among the studied variables (reaction temperature, molar ratio of oil/alcohol, enzyme concentration and ratio v/v oil/hexane), only the enzymatic concentration as a significant variable and indicated a concentration of 10% of immobilized hyphae as an optimum point. However, experimentally speaking, the best biodiesel yield was 15.95%, using 7.75% immobilized hyphae, at 31.25°C, with 1:7.5 oil/alcohol molar ratio... (Complete abstract click electronic access below)
Descrição
Palavras-chave
Microbiologia industrial, Transesterificação, Lipase, Enzimas de fungos - Aplicações industriais, Ethylic biodiesel
Como citar
BORGES, Janaina Pires. Produção de lipases por fungos termofílicos e mesofílicos e uso na produção de biodiesel etílico. 2012. 78 f. Dissertação (mestrado) - Universidade Estadual Paulista, Instituto de Biociências, Letras e Ciências Exatas, 2012.