Produção de ácidos graxos por linhagens de Mucor spp. isoladas de solo de área de cerrado, município de Corumbataí, Estado de São Paulo
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2006-10-26
Autores
Wellbaum, Christian [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
Na área de cerrado, município de Corumbatai, SP, foram isoladas 267 linhagens de fungos filamentosos em amostras de solo, coletadas no inverno (2002) e no verão (2003), pelos métodos da Técnica da placa-de-solo e Técnica da diluição em série. Destas, 12 linhagens foram consideradas potenciais produtoras de ácidos Á-linolenico (AGL) e araquidônico (AA) por pertencerem a ordem Mucorales. Foram realizados ensaios preliminares para avaliar a eficiência de diversos protocolos de extração de ácidos graxos dos fungos. Foram testadas modificações de metodologias tradicionais, adicionando uma ou duas vezes a solução extratora de metanol-cloroformio-água (2,0:1,0:0,8 v/v) a biomassa úmida, congelada, liofilizada, seca e/ou macerada. De acordo com os resultados obtidos, optou-se por utilizar a biomassa liofilizada, tratada duas vezes com a solução extratora. Os extratos resultantes foram transesterificados e analisados por cromatografia gasosa. Algumas linhagens foram cultivadas em erlenmeyeres de 125mL e 250mL, contendo respectivamente 50mL e 100mL de meio de cultura líquido de extrato de malte a 2%, durante 6 dias sob agitação, a 25C. Foi observado que a linhagem MX produziu maior quantidade de AGL, apresentando médias de 61,9mg.L-1e 55,7mgEL-1, respectivamente, em frascos de 250mL e 125mL. Maior produção de AA foi verificada na linhagem M132, cuja média foi de 0,3mgEL-1. As linhagens MX, M131 e M132 foram também cultivadas em frascos de 125mL contendo 50mL de meio líquido de extrato-de-malte suplementado com óleo de girassol a 2% e 1% de extrato de levedura. Nestes, as maiores quantidades médias de AGL, 201,62mgEL-1, e de AA, 3,12mgEL-1, foram observadas no cultivo da linhagem M131, sendo estes valores 1.311,15% e 1011,37% maiores em relação aqueles verificados em meio de extrato de malte.
In the savannah area, municipal district of Corumbatai, state of Sao Paulo, 267 fungi strains were isolated in soil samples collected, in the winter (2002) and in the summer (2003), with the gsoil plate methodh and serial dilution method. Twelve strains were considered potential producers of Á-linolenic acid (GLA) and arachidonic acid (AA) for belonging to the order Mucorales. Preliminary experiments were accomplished to evaluate the efficiency of several protocols of extraction of fatty acids from fungi. Modifications of traditional methodologies were tested, adding one or twice the solvent solution of methanol-chloroform-water (2.0:1.0:0.8 v/v) to the wet, frozen, freeze-dried, dried and/or disrupted biomass. In agreement with the obtained results, the biomass was freeze-dried and treated twice with the solvent solution. The resulting extracts were transesterified and analized by gas cromatography Some strains were cultivated in shake flasks of 125mL and 250mL, containing respectively 50mL and 100mL of liquid malt extract culture media (2%), for 6 days under agitation, at 25C. It was observed that the strain MX produced larger amounts of GLA, mean of 61.9mgEL-1 and 55.7mgEL-1, respectively, in shake flasks of 250mL and 125mL. Higher production of AA was verified in the strain M132, mean of 0.3mgEL-1. The srains MX, M131 and M132 were also cultivated in shake flasks of 125mL containing 50mL of liquid malt extract media enhanced with sunflower oil (2%) and yeast extract (1%). In these the largest amounts of GLA, mean of 201.62mgEL-1, and of AA, mean of 3.12mgEL-1, were observed in the cultivation of the strain M131, these values being 1,311.15% and 1,011.37% larger in relation to those verified in same strain cultivated in malt extract media. The three strains were grown in Bioreactor with capacity of 2L containing 1L of malt extract media (2%) added or not with sunflower oil (2%).
In the savannah area, municipal district of Corumbatai, state of Sao Paulo, 267 fungi strains were isolated in soil samples collected, in the winter (2002) and in the summer (2003), with the gsoil plate methodh and serial dilution method. Twelve strains were considered potential producers of Á-linolenic acid (GLA) and arachidonic acid (AA) for belonging to the order Mucorales. Preliminary experiments were accomplished to evaluate the efficiency of several protocols of extraction of fatty acids from fungi. Modifications of traditional methodologies were tested, adding one or twice the solvent solution of methanol-chloroform-water (2.0:1.0:0.8 v/v) to the wet, frozen, freeze-dried, dried and/or disrupted biomass. In agreement with the obtained results, the biomass was freeze-dried and treated twice with the solvent solution. The resulting extracts were transesterified and analized by gas cromatography Some strains were cultivated in shake flasks of 125mL and 250mL, containing respectively 50mL and 100mL of liquid malt extract culture media (2%), for 6 days under agitation, at 25C. It was observed that the strain MX produced larger amounts of GLA, mean of 61.9mgEL-1 and 55.7mgEL-1, respectively, in shake flasks of 250mL and 125mL. Higher production of AA was verified in the strain M132, mean of 0.3mgEL-1. The srains MX, M131 and M132 were also cultivated in shake flasks of 125mL containing 50mL of liquid malt extract media enhanced with sunflower oil (2%) and yeast extract (1%). In these the largest amounts of GLA, mean of 201.62mgEL-1, and of AA, mean of 3.12mgEL-1, were observed in the cultivation of the strain M131, these values being 1,311.15% and 1,011.37% larger in relation to those verified in same strain cultivated in malt extract media. The three strains were grown in Bioreactor with capacity of 2L containing 1L of malt extract media (2%) added or not with sunflower oil (2%).
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Palavras-chave
Fungos, Cerrados, Ácido araquidônico, Savannah, Fungi
Como citar
WELLBAUM, Christian. Produção de ácidos graxos por linhagens de Mucor spp. isoladas de solo de área de cerrado, município de Corumbataí, Estado de São Paulo. 2006. x, 73 f. Tese (doutorado) - Universidade Estadual Paulista, Instituto de Biociências, 2006.