Production, Purification, and Characterization of a Major Penicillium glabrum Xylanase Using Brewer's Spent Grain as Substrate

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 thi...

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Main Authors: Adriana Knob, Susan Michelz Beitel, Diana Fortkamp, César Rafael Fanchini Terrasan, Alex Fernando de Almeida
Format: Article
Language:English
Published: Hindawi Limited 2013-01-01
Series:BioMed Research International
Online Access:http://dx.doi.org/10.1155/2013/728735
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spelling doaj-0075af0a05e54188ac73685ab9f01b572020-11-24T23:02:58ZengHindawi LimitedBioMed Research International2314-61332314-61412013-01-01201310.1155/2013/728735728735Production, Purification, and Characterization of a Major Penicillium glabrum Xylanase Using Brewer's Spent Grain as SubstrateAdriana Knob0Susan Michelz Beitel1Diana Fortkamp2César Rafael Fanchini Terrasan3Alex Fernando de Almeida4Department of Biological Sciences, Midwest State University, Camargo Varela de Sá Street 03, 85040-080 Guarapuava, PR, BrazilDepartment of Biological Sciences, Midwest State University, Camargo Varela de Sá Street 03, 85040-080 Guarapuava, PR, BrazilDepartment of Biological Sciences, Midwest State University, Camargo Varela de Sá Street 03, 85040-080 Guarapuava, PR, BrazilDepartment of Chemical Engineering, Federal University of São Carlos, Rodovia Washington Luís, km 235, SP-310, 13565-905 São Carlos, SP, BrazilDepartment of Biochemistry and Microbiology, São Paulo State University, 24-A Avenue 1515, 13506-900 Rio Claro, SP, BrazilIn 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.http://dx.doi.org/10.1155/2013/728735
collection DOAJ
language English
format Article
sources DOAJ
author Adriana Knob
Susan Michelz Beitel
Diana Fortkamp
César Rafael Fanchini Terrasan
Alex Fernando de Almeida
spellingShingle Adriana Knob
Susan Michelz Beitel
Diana Fortkamp
César Rafael Fanchini Terrasan
Alex Fernando de Almeida
Production, Purification, and Characterization of a Major Penicillium glabrum Xylanase Using Brewer's Spent Grain as Substrate
BioMed Research International
author_facet Adriana Knob
Susan Michelz Beitel
Diana Fortkamp
César Rafael Fanchini Terrasan
Alex Fernando de Almeida
author_sort Adriana Knob
title Production, Purification, and Characterization of a Major Penicillium glabrum Xylanase Using Brewer's Spent Grain as Substrate
title_short Production, Purification, and Characterization of a Major Penicillium glabrum Xylanase Using Brewer's Spent Grain as Substrate
title_full Production, Purification, and Characterization of a Major Penicillium glabrum Xylanase Using Brewer's Spent Grain as Substrate
title_fullStr Production, Purification, and Characterization of a Major Penicillium glabrum Xylanase Using Brewer's Spent Grain as Substrate
title_full_unstemmed Production, Purification, and Characterization of a Major Penicillium glabrum Xylanase Using Brewer's Spent Grain as Substrate
title_sort production, purification, and characterization of a major penicillium glabrum xylanase using brewer's spent grain as substrate
publisher Hindawi Limited
series BioMed Research International
issn 2314-6133
2314-6141
publishDate 2013-01-01
description 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.
url http://dx.doi.org/10.1155/2013/728735
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