This study aimed to verify the hydrolysis of starch raw cassava using a glucoamylase from Aspergillus awamori expressed in Saccharomyces cerevisiae. The glucoamylase activity test plate showed that S. cerevisiae recombinant is able to degrade cassava starch without the needs for either acid pretreatment or cooking. The recombinant yeast was able to form 2.7 g / L of biomass and degrades cassava starch up to 90%, releasing a large amount of glucose to the culture medium. The N-deglycosylation assay showed that the recombinant glucoamylase (75 kDa) has 10% carbohydrate in their structure compared with enzyme treated (65 kDa). Our preliminary results showed that recombinant yeast was able to produce 4.22 g/L in 96 hours fermentation. The results of starch hydrolysis shown that S. cerevisiae recombinant has a great potential for producing ethanol from cassava starch without the need for pretreatment of the substrate.
RESUMOEsse trabalho teve como objetivo verificar a hidrólise do amido bruto de mandioca, usando a glicoamilase de Aspergillus awamori expressa em Saccharomyces cerevisiae. O teste de atividade de glicoamilase em placa mostrou que a S. cerevisiae recombinante é capaz de degradar o amido de mandioca sem a necessidade de pré-tratamento acido ou cozimento. A levedura recombinante foi capaz de formar 2,7 g/L de biomassa que degradou 90% do amido, liberando uma grande quantidade de glicose para o meio de cultura. O ensaio de deglicosilação mostrou que a glicoamilase recombinante (75 kDa) possui 10% de carboidrato na sua estrutura comparado com a enzima deglicosilada (65 kDa). Nossos resultados preliminares mostraram que a levedura recombinante produziu 4,22 g/L de bioetanol em 96 horas de fermentação. Os resultados da hidrólise do amido mostraram que a S. cerevisiae recombinante utilizada nesse trabalho tem um grande potencial de produzir etanol a partir de amido de mandioca sem a necessidade de um pré-tratamento do substrato.
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