title: Production, purification, and characterization of a major Penicillium glabrum xylanase using Brewer's spent grain as substrate.
abstract: 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 Mn(2+) and the reducing agents β -mercaptoethanol and DTT enhanced xylanase activity, while the ions Hg(2+), Zn(2+), and Cu(2+) 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.
authors.affiliation: Department of Biological Sciences, Midwest State University, Camargo Varela de Sá Street 03, 85040-080 Guarapuava, PR, Brazil.
authors.forename: Adriana
authors.initials: A
authors.lastname: Knob
authors.forename: Susan Michelz
authors.initials: SM
authors.lastname: Beitel
authors.forename: Diana
authors.initials: D
authors.lastname: Fortkamp
authors.forename: César Rafael Fanchini
authors.initials: CR
authors.lastname: Terrasan
authors.forename: Alex Fernando
authors.initials: AF
authors.lastname: de Almeida
mesh_terms: D002241:Carbohydrates / Q000494:pharmacology; D002244:Carbon / Q000494:pharmacology; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D043364:Endo-1,4-beta Xylanases / Q000096:biosynthesis* / Q000302:isolation & purification* / Q000378:metabolism; D004795:Enzyme Stability / Q000187:drug effects; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D007220:Industrial Waste / Q000032:analysis*; D010407:Penicillium / Q000201:enzymology*; D013379:Substrate Specificity / Q000187:drug effects; D013696:Temperature; D013997:Time Factors; D014866:Waste Products / Q000032:analysis*
chemical_list: D002241:Carbohydrates; D007220:Industrial Waste; D014866:Waste Products; D002244:Carbon; D043364:Endo-1,4-beta Xylanases