Production, purification, and characterization of a major penicillium glabrum xylanase using brewer's spent grain as substrate

• Autor(a) principal: Knob, Adriana
• Data de Publicação: 2013
• Outros Autores: Beitel, Susan Michelz, Fortkamp, Diana, Terrasan, César Rafael Fanchini, Almeida, Alex Fernando de [UNESP]
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UNESP
• Texto Completo: http://dx.doi.org/10.1155/2013/728735; http://hdl.handle.net/11449/75641
• topic: ammonium sulfate; cupric ion; magnesium ion; mercaptoethanol; mercury; xylan endo 1,3 beta xylosidase; zinc ion; controlled study; culture medium; enzyme activity; enzyme analysis; enzyme purification; enzyme stability; enzyme substrate; enzyme synthesis; fungus culture; fungus growth; grain; ion exclusion chromatography; molecular size; nonhuman; Penicillium; Penicillium glabrum; pH; polyacrylamide gel electrophoresis; process optimization

In recent decades, xylanases have been used in many processing industries. This study describes the xylanase production by Penicillium glabrum using brewer's spent grain as substrate. Additionally, this is the first work that reports the purification and characterization of a xylanase using this agroindustrial waste. Optimal production was obtained when P. glabrum was grown in liquid medium in pH 5.5, at 25 °C, under stationary condition for six days. The xylanase from P. glabrum was purified to homogeneity by a rapid and inexpensive procedure, using ammonium sulfate fractionation and molecular exclusion chromatography. SDS-PAGE analysis revealed one band with estimated molecular mass of 18.36 kDa. The optimum activity was observed at 60 °C, in pH 3.0. The enzyme was very stable at 50 °C, and high pH stability was verified from pH 2.5 to 5.0. The ion Mn2+ and the reducing agents β-mercaptoethanol and DTT enhanced xylanase activity, while the ions Hg2+, Zn2+, and Cu2+ as well as the detergent SDS were strong inhibitors of the enzyme. The use of brewer's spent grain as substrate for xylanase production cannot only add value and decrease the amount of this waste but also reduce the xylanase production cost. © 2013 Adriana Knob et al.