Síntese e caracterização da lipase de Aspergillus oryzae
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Data
2023-05-18
Autores
Barbosa, Izabela
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Universidade Estadual Paulista (Unesp)
Resumo
Tendo em vista que enzimas possuem grande importância industrial, esse trabalho
teve como objetivo estudar a síntese e caracterização de lipase de Aspergillus oryzae,
uma vez que são biocatalisadores versáteis e com grande potencial em diversas
aplicações. Para isso, foi necessário avaliar as características morfológicas do fungo
e estudar algumas condições de cultivo, como o pH, tempo de fermentação e
composição do meio de cultivo em relação as fontes de carbono e nitrogênio, bem
como caracterizar a lipase quanto ao pH e a temperatura ideais. Na primeira etapa, o
cultivo do Aspergillus oryzae em placas de Petri possibilitou a caracterização
qualitativa do mesmo, que cresceu formando colônias de coloração amarelo esverdeada. Nas etapas seguintes, foram realizados planejamentos experimentais de
forma a aprimorar o processo e otimizar o tempo e custos de análise. Com base no
delineamento composto central (DCC) 2
3
realizado em shaker, o melhor tempo de
fermentação foi em 120h, com pH 6, 4,5 g/L de sulfato de amônio e 14 g/L de sacarose,
obtendo-se lipase com atividade hidrolítica de 94,33 ± 5,51 U/mL. Por fim, através de
um delineamento composto central rotacional (DCCR) de 2 fatores realizado em
shaker, a lipase apresentou pH ideal de 7,5 e temperatura ideal de 30ºC. Diante disso,
este trabalho permitiu a definição de melhores parâmetros para síntese da lipase de
Aspergillus oryzae e sua caracterização, recomendando-se que trabalhos futuros
estudem a especificidade da enzima e suas possíveis aplicações.
Considering that enzymes are of great industrial importance, this work aimed to study the synthesis and characterization of lipase from Aspergillus oryzae, as they are versatile biocatalysts with great potential in various applications. For this, it was necessary to evaluate the morphological characteristics of the fungus and study culture parameters, such as pH, fermentation time and composition of the production medium in relation to carbon and nitrogen sources, as well as to characterize the lipase as to its ideal pH and temperature. In the first stage, the cultivation of Aspergillus oryzae in Petri dishes allowed the qualitative characterization of it, which grew forming colonies of greenish yellow color. In the following stages, experimental designs were made in order to improve the process and optimize time and cost of the analysis. Based on the central composite design (CCD) 2 3 performed in a shaker, the best fermentation time was 120 hours, with pH 6, 4.5 g/L of ammonium sulfate and 14 g/L of sucrose, obtaining lipase with hydrolytic activity of 94.33 ± 5.51 U/mL. Finally, through a central composite rotational design (CCRD) with two factors performed in a shaker, the lipase had an ideal pH of 7.5 and an ideal temperature of 30ºC. Thus, this work allowed the definition of the best parameters for the synthesis of lipase from Aspergillus oryzae and its characterization, recommending that future studies investigate the enzyme’s specificity and its potential applications.
Considering that enzymes are of great industrial importance, this work aimed to study the synthesis and characterization of lipase from Aspergillus oryzae, as they are versatile biocatalysts with great potential in various applications. For this, it was necessary to evaluate the morphological characteristics of the fungus and study culture parameters, such as pH, fermentation time and composition of the production medium in relation to carbon and nitrogen sources, as well as to characterize the lipase as to its ideal pH and temperature. In the first stage, the cultivation of Aspergillus oryzae in Petri dishes allowed the qualitative characterization of it, which grew forming colonies of greenish yellow color. In the following stages, experimental designs were made in order to improve the process and optimize time and cost of the analysis. Based on the central composite design (CCD) 2 3 performed in a shaker, the best fermentation time was 120 hours, with pH 6, 4.5 g/L of ammonium sulfate and 14 g/L of sucrose, obtaining lipase with hydrolytic activity of 94.33 ± 5.51 U/mL. Finally, through a central composite rotational design (CCRD) with two factors performed in a shaker, the lipase had an ideal pH of 7.5 and an ideal temperature of 30ºC. Thus, this work allowed the definition of the best parameters for the synthesis of lipase from Aspergillus oryzae and its characterization, recommending that future studies investigate the enzyme’s specificity and its potential applications.
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Palavras-chave
Lipase, Aspergillus oryzae, Sacarose, Sucrose, Sulfato de amônio, Ammonium sulphate