One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ash

Detalhes bibliográficos
Autor(a) principal: Sanghi,Ashwani
Data de Publicação: 2010
Outros Autores: Garg,Neelam, Gupta,V.K., Mittal,Ashwani, Kuhad,R.C.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Journal of Microbiology
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822010000200029
Resumo: The present study describes the one-step purification and characterization of an extracellular cellulase-free xylanase from a newly isolated alkalophilic and moderately thermophilic strain of Bacillus subtilis ASH. Xylanase was purified to homogeneity by 10.5-fold with ~43% recovery using ion-exchange chromatography through CM-Sephadex C-50. The purified enzyme revealed a single band on SDS-PAGE gel with a molecular mass of 23 kDa. It showed an optimum pH at 7.0 and was stable over the pH range 6.0-9.0. The optimum temperature for enzyme activity was 55 ºC. The purified xylanase did not lose any activity up to 45 ºC, however, it retained 80% and 51% of its activity after pre-incubation at 55 ºC and 60 ºC, respectively. The enzyme obeyed Michaelis-Menton kinetics towards birch wood xylan with apparent Km 3.33 mg/ml and Vmax 100 IU/ml. The enzyme was strongly inhibited by Hg2+ and Cu2+ while enhanced by Co2+ and Mn2+. The purified enzyme could be stored at 4 ºC for six weeks without any loss of catalytic activity. The faster and economical purification of the cellulase-free xylanase from B. subtilis ASH by one-step procedure together with its appreciable stability at high temperature and alkaline pH makes it potentially effective for industrial applications.
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spelling One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ashAlkalophilicBacillus subtilisPurificationXylanaseThe present study describes the one-step purification and characterization of an extracellular cellulase-free xylanase from a newly isolated alkalophilic and moderately thermophilic strain of Bacillus subtilis ASH. Xylanase was purified to homogeneity by 10.5-fold with ~43% recovery using ion-exchange chromatography through CM-Sephadex C-50. The purified enzyme revealed a single band on SDS-PAGE gel with a molecular mass of 23 kDa. It showed an optimum pH at 7.0 and was stable over the pH range 6.0-9.0. The optimum temperature for enzyme activity was 55 ºC. The purified xylanase did not lose any activity up to 45 ºC, however, it retained 80% and 51% of its activity after pre-incubation at 55 ºC and 60 ºC, respectively. The enzyme obeyed Michaelis-Menton kinetics towards birch wood xylan with apparent Km 3.33 mg/ml and Vmax 100 IU/ml. The enzyme was strongly inhibited by Hg2+ and Cu2+ while enhanced by Co2+ and Mn2+. The purified enzyme could be stored at 4 ºC for six weeks without any loss of catalytic activity. The faster and economical purification of the cellulase-free xylanase from B. subtilis ASH by one-step procedure together with its appreciable stability at high temperature and alkaline pH makes it potentially effective for industrial applications.Sociedade Brasileira de Microbiologia2010-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822010000200029Brazilian Journal of Microbiology v.41 n.2 2010reponame:Brazilian Journal of Microbiologyinstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1590/S1517-83822010000200029info:eu-repo/semantics/openAccessSanghi,AshwaniGarg,NeelamGupta,V.K.Mittal,AshwaniKuhad,R.C.eng2010-04-16T00:00:00Zoai:scielo:S1517-83822010000200029Revistahttps://www.scielo.br/j/bjm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||mbmartin@usp.br1678-44051517-8382opendoar:2010-04-16T00:00Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)false
dc.title.none.fl_str_mv One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ash
title One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ash
spellingShingle One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ash
Sanghi,Ashwani
Alkalophilic
Bacillus subtilis
Purification
Xylanase
title_short One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ash
title_full One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ash
title_fullStr One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ash
title_full_unstemmed One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ash
title_sort One-step purification and characterization of cellulase-free xylanase produced by alkalophilic Bacillus subtilis ash
author Sanghi,Ashwani
author_facet Sanghi,Ashwani
Garg,Neelam
Gupta,V.K.
Mittal,Ashwani
Kuhad,R.C.
author_role author
author2 Garg,Neelam
Gupta,V.K.
Mittal,Ashwani
Kuhad,R.C.
author2_role author
author
author
author
dc.contributor.author.fl_str_mv Sanghi,Ashwani
Garg,Neelam
Gupta,V.K.
Mittal,Ashwani
Kuhad,R.C.
dc.subject.por.fl_str_mv Alkalophilic
Bacillus subtilis
Purification
Xylanase
topic Alkalophilic
Bacillus subtilis
Purification
Xylanase
description The present study describes the one-step purification and characterization of an extracellular cellulase-free xylanase from a newly isolated alkalophilic and moderately thermophilic strain of Bacillus subtilis ASH. Xylanase was purified to homogeneity by 10.5-fold with ~43% recovery using ion-exchange chromatography through CM-Sephadex C-50. The purified enzyme revealed a single band on SDS-PAGE gel with a molecular mass of 23 kDa. It showed an optimum pH at 7.0 and was stable over the pH range 6.0-9.0. The optimum temperature for enzyme activity was 55 ºC. The purified xylanase did not lose any activity up to 45 ºC, however, it retained 80% and 51% of its activity after pre-incubation at 55 ºC and 60 ºC, respectively. The enzyme obeyed Michaelis-Menton kinetics towards birch wood xylan with apparent Km 3.33 mg/ml and Vmax 100 IU/ml. The enzyme was strongly inhibited by Hg2+ and Cu2+ while enhanced by Co2+ and Mn2+. The purified enzyme could be stored at 4 ºC for six weeks without any loss of catalytic activity. The faster and economical purification of the cellulase-free xylanase from B. subtilis ASH by one-step procedure together with its appreciable stability at high temperature and alkaline pH makes it potentially effective for industrial applications.
publishDate 2010
dc.date.none.fl_str_mv 2010-06-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822010000200029
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S1517-83822010000200029
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S1517-83822010000200029
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv text/html
dc.publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
dc.source.none.fl_str_mv Brazilian Journal of Microbiology v.41 n.2 2010
reponame:Brazilian Journal of Microbiology
instname:Sociedade Brasileira de Microbiologia (SBM)
instacron:SBM
instname_str Sociedade Brasileira de Microbiologia (SBM)
instacron_str SBM
institution SBM
reponame_str Brazilian Journal of Microbiology
collection Brazilian Journal of Microbiology
repository.name.fl_str_mv Brazilian Journal of Microbiology - Sociedade Brasileira de Microbiologia (SBM)
repository.mail.fl_str_mv bjm@sbmicrobiologia.org.br||mbmartin@usp.br
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