title: Characterization of β -Glucosidase Produced by Aspergillus niger under Solid-State Fermentation and Partially Purified Using MANAE-Agarose.
abstract: β -Glucosidase (BGL) is a hydrolytic enzyme with specificity for a wide variety of glycoside substrates, being an enzyme with a large range of biotechnological applications. However, enzyme properties can be different depending both on the microorganism and the cultivation procedure employed. Therefore, in order to explore potential biocatalytical applications of novel enzymes, their characterization is essential. In this work, a BGL synthesized by a selected strain of Aspergillus niger cultivated under solid-state fermentation (SSF) was partially purified and fully characterized in terms of optimum pH, temperature, and thermostability. The single-step purification using MANAE-agarose in a chromatographic column yielded an enzyme solution with specific activity (17.1 IU/mg protein) adequate for the characterization procedures. Electrophoresis SDS-PAGE and size-exclusion chromatography analysis resulted in an estimated molecular mass of 60 kDa. Higher enzyme activities were found in the range between 40 and 65°C and between pH 4 and 5.5, indicating an interesting characteristic for application in the hydrolysis of lignocellulosic biomass for biofuels production. Thermostability studies of purified BGL resulted in half-lives at 37°C of 56.3 h and at 50°C of 5.4 h. These results provide support for further studies of this enzyme towards revealing its potential biotechnological applications.
authors.affiliation: Departamento de Engenharia Química, Universidade Federal de São Carlos, Rodovia Washington Luiz km 235, 13565-905 São Carlos, SP, Brazil ; Embrapa Instrumentação, Rua XV de Novembro 1452, 13560-970 São Carlos, São Paulo, SP, Brazil.
authors.forename: Anderson
authors.identifier: 0000-0003-3927-4538
authors.initials: A
authors.lastname: Baraldo Junior
authors.affiliation: Departamento de Engenharia Química, Universidade Federal de São Carlos, Rodovia Washington Luiz km 235, 13565-905 São Carlos, SP, Brazil.
authors.forename: Diogo G
authors.initials: DG
authors.lastname: Borges
authors.affiliation: Departamento de Engenharia Química, Universidade Federal de São Carlos, Rodovia Washington Luiz km 235, 13565-905 São Carlos, SP, Brazil.
authors.forename: Paulo W
authors.initials: PW
authors.lastname: Tardioli
authors.affiliation: Departamento de Engenharia Química, Universidade Federal de São Carlos, Rodovia Washington Luiz km 235, 13565-905 São Carlos, SP, Brazil ; Embrapa Instrumentação, Rua XV de Novembro 1452, 13560-970 São Carlos, São Paulo, SP, Brazil.
authors.forename: Cristiane S
authors.initials: CS
authors.lastname: Farinas