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Production of 2-phenylethanol by biotransformation of 2-phenylnitroethane using different fungi strains
Darlisson de Alexandria Santos
Universidade Federal do Sul e Sudeste do Pará
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2-Phenylethanol is a well know alcohol that is characterized by its rose-like odor, being used as component in fragrance, cosmetic, and food industries1. It occurs widely in nature, mainly in essential oils extracted from rose, jasmine, carnation, and hyacinths2. However, their low concentration makes the recovery complex and expensive1,2. 2-Phenylethanol was also described as showing antimicrobial and antityrosinase activities, which aggregates more attention to its production3. 2-Phenylnitroethane, used in this work as substrate for 2-phenylethanol, is the main component of Aniba canelilla essential oil, corresponding to approximately 80% of the total amount. Then, it was used as substrate without any previous purification. To produce 2-phenylethanol through biotransformation of 2-phenylnitroethane, four different fungi were applied: Aspergillus flavus and Rigidoporus lineatus (isolated from copper mining residue from Mina do Sossego, Canaã dos Carajás, Pará, Brazil), Lasiodiplodia caatinguensis (phytopathogenic fungus from Citrus sinensis), and Colletotrichum sp. (phytopathogenic fungus from Euterpe oleracea). The fungi were cultivated in Erlenmeyer flasks (250 mL) containing 100 mL of malt extract solution (2%) for seven days under orbital agitation (132 rpm) at 30 ºC. Then, 50 mg of A. canelilla essential oil were diluted in 100 µL of dimethylsulfoxide (DMSO) and added to each fungi cultivation. Each experiment had its control by fungi cultivation without the essential oil addition. Every day (1 - 7 days), aliquots (2 mL) were removed from each experiment and extracted using ethyl acetate (2 mL) and analyzed by GC-MS in order to determine the percentage of 2-phenylnitroethane biotransformation in 2-phenylethanol. After 7 days, it was observed that the two fungi (A. flavus, and L. caatinguensis) performed the complete biotransformation of 2-phenylnitroethane in 2-phenylethanol, with special attention to L. caatinguensis that achieved the complete biotransformation after two days of reaction. The other two fungi (R. lineatus, and Colletotrichum sp.) partially performed the biotransformation, but Colletotrichum sp. also produced a not-identified compound, which will the object of future studies. Therefore, it was possible to assign that A. flavus and L. caatinguensis fungi are very promisors in 2-phenylethanol production using 2-phenylnitroethane as substrate and can be the object of study in the next steps of this work, that includes the improvement of the reactional conditions and the production of 2-phenylethanol using other substrates such as hydrolyzed lignin and cinnamyl alcohol.
Emmanuel Dias da Silva
Olá, Darlisson Santos
Boa apresentação e bom trabalho, parabéns. Minhas dúvidas: Qual a quantidade de material de partida e qual a massa/volume/mmol de produto final (álcool)? Vcs pensam em investir em novos fungos? Vi que a diferença no tempo de reação foi de 7d para um fungo e 2d para outro e, por isso, pergunto: vcs fizeram varreduras de temperatura/solvente/quantidade de substrato? Abraço
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Darlisson de Alexandria Santos
Olá, muito obrigado pelas perguntas.
Na verdade este trabalho é fruto de um screening inicial e não tivemos tempo de estudar outros parâmetros devido à pandemia. Faz parte dos objetivos variarmos a concentração de substrato, temperatura, pH e até mudar a concentração do meio de cultura. Feitas estas variações, então determinaremos a produtividade, em um estudo cinético ao longo do tempo de fermentação.
Tenho a impressão que outros substratos com estrutura química parecida possam ser usados também por estes microrganismos para a produção do composto de interesse. Por isso, vamos avaliar a lignina hidrolisada como um desses prováveis substratos.