title: Production, purification and characterization of a thermostable β-1,3-glucanase produced by Moniliophthora perniciosa.
abstract: The enzyme glucanase from Moniliophthora perniciosa was produced in liquid medium and purified from the culture supernatant. A multivariate statistical approach (Response Surface Methodology - RSM) was employed to evaluate the effect of variables, including inducer (yeast extract) and fermentation time, on secreted glucanase activities M. perniciosa detected in the culture medium. The crude enzyme present in the supernatant was purified in two steps: precipitation with ammonium sulfate (70%) and gel filtration chromatography on Sephacryl S-200. The best inducer and fermentation time for glucanase activities were 5.9 g L(-1) and 13 days, respectively. The results revealed three different isoforms (GLUI, GLUII and GLUIII) with purification factors of 4.33, 1.86 and 3.03, respectively. The partially purified enzymatic extract showed an optimum pH of 5.0 and an optimum temperature of 40°C. The enzymatic activity increased in the presence of KCl at all concentrations studied. The glucanase activity was highest in the presence of 0.2 M NaCl. The enzyme showed high thermal stability, losing only 10.20% of its specific activity after 40 minutes of incubation at 90°C. A purified enzyme with relatively good thermostability that is stable at low pH might be used in future industrial applications.
authors.affiliation: Departamento de Saúde, Universidade Estadual de Feira de Santana, BA, Brasil.
authors.forename: Amanda R
authors.initials: AR
authors.lastname: Sena
authors.forename: Gildomar L V
authors.initials: GL
authors.lastname: Júnior
authors.forename: Aristóteles
authors.initials: A
authors.lastname: Góes Neto
authors.forename: Alex G
authors.initials: AG
authors.lastname: Taranto
authors.forename: Carlos P
authors.initials: CP
authors.lastname: Pirovani
authors.forename: Júlio C M
authors.initials: JC
authors.lastname: Cascardo
authors.forename: Russolina B
authors.initials: RB
authors.lastname: Zingali
authors.forename: Marcos A
authors.initials: MA
authors.lastname: Bezerra
authors.forename: Sandra A
authors.initials: SA
authors.lastname: Assis
mesh_terms: D000363:Agaricales / Q000201:enzymology*; D002850:Chromatography, Gel; D004795:Enzyme Stability; D005285:Fermentation; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D004693:Glucan Endo-1,3-beta-D-Glucosidase