Production, characterization and purification of chitosanase from Bacillus cereus
Autor(a) principal: | |
---|---|
Data de Publicação: | 1999 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Brazilian Journal of Chemical Engineering |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0104-66321999000200011 |
Resumo: | A culture medium for a wild strain of Bacillus cereus was developed for chitosanase production by using an experimental design. The factors having the strongest influence on chitosanase production were ammonium sulfate concentration, aeration, pH and the interaction between the first two parameters. Optimal conditions for chitosan hydrolysis were pH 5.8 and 54 oC; however, hydrolysis activity drastically decreased at pH 7.0. The enzyme was purified (single-electrophoretic band) by partitioning in an aqueous two-phase system (ATPS), followed by cation-exchange chromatography with a 66% yield. Chitosanase was mainly collected in the top phase (K = 129) of a 22% PEG 1,500, 13% phosphate (pH = 5.8) and 12% NaCl (w/w) solution, and the main protein contaminants were evenly distributed between the phases (K = 1.07). The apparent molecular weight and the isoelectric point of the chitosanase, determined by SDS-PAGE electrophoresis and by isoelectric focalization, were 47 kDa and 8.8, respectively. |
id |
ABEQ-1_ef4bccf477320248a9d230e358840a66 |
---|---|
oai_identifier_str |
oai:scielo:S0104-66321999000200011 |
network_acronym_str |
ABEQ-1 |
network_name_str |
Brazilian Journal of Chemical Engineering |
repository_id_str |
|
spelling |
Production, characterization and purification of chitosanase from Bacillus cereusChitosanaseBacillus sp.factorial experimental designaqueous two-phase systemsA culture medium for a wild strain of Bacillus cereus was developed for chitosanase production by using an experimental design. The factors having the strongest influence on chitosanase production were ammonium sulfate concentration, aeration, pH and the interaction between the first two parameters. Optimal conditions for chitosan hydrolysis were pH 5.8 and 54 oC; however, hydrolysis activity drastically decreased at pH 7.0. The enzyme was purified (single-electrophoretic band) by partitioning in an aqueous two-phase system (ATPS), followed by cation-exchange chromatography with a 66% yield. Chitosanase was mainly collected in the top phase (K = 129) of a 22% PEG 1,500, 13% phosphate (pH = 5.8) and 12% NaCl (w/w) solution, and the main protein contaminants were evenly distributed between the phases (K = 1.07). The apparent molecular weight and the isoelectric point of the chitosanase, determined by SDS-PAGE electrophoresis and by isoelectric focalization, were 47 kDa and 8.8, respectively.Brazilian Society of Chemical Engineering1999-06-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0104-66321999000200011Brazilian Journal of Chemical Engineering v.16 n.2 1999reponame:Brazilian Journal of Chemical Engineeringinstname:Associação Brasileira de Engenharia Química (ABEQ)instacron:ABEQ10.1590/S0104-66321999000200011info:eu-repo/semantics/openAccessPIZA,F. A. T.SILOTO,A. P.CARVALHO,C. V.FRANCO,T.T.eng1999-09-15T00:00:00Zoai:scielo:S0104-66321999000200011Revistahttps://www.scielo.br/j/bjce/https://old.scielo.br/oai/scielo-oai.phprgiudici@usp.br||rgiudici@usp.br1678-43830104-6632opendoar:1999-09-15T00:00Brazilian Journal of Chemical Engineering - Associação Brasileira de Engenharia Química (ABEQ)false |
dc.title.none.fl_str_mv |
Production, characterization and purification of chitosanase from Bacillus cereus |
title |
Production, characterization and purification of chitosanase from Bacillus cereus |
spellingShingle |
Production, characterization and purification of chitosanase from Bacillus cereus PIZA,F. A. T. Chitosanase Bacillus sp. factorial experimental design aqueous two-phase systems |
title_short |
Production, characterization and purification of chitosanase from Bacillus cereus |
title_full |
Production, characterization and purification of chitosanase from Bacillus cereus |
title_fullStr |
Production, characterization and purification of chitosanase from Bacillus cereus |
title_full_unstemmed |
Production, characterization and purification of chitosanase from Bacillus cereus |
title_sort |
Production, characterization and purification of chitosanase from Bacillus cereus |
author |
PIZA,F. A. T. |
author_facet |
PIZA,F. A. T. SILOTO,A. P. CARVALHO,C. V. FRANCO,T.T. |
author_role |
author |
author2 |
SILOTO,A. P. CARVALHO,C. V. FRANCO,T.T. |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
PIZA,F. A. T. SILOTO,A. P. CARVALHO,C. V. FRANCO,T.T. |
dc.subject.por.fl_str_mv |
Chitosanase Bacillus sp. factorial experimental design aqueous two-phase systems |
topic |
Chitosanase Bacillus sp. factorial experimental design aqueous two-phase systems |
description |
A culture medium for a wild strain of Bacillus cereus was developed for chitosanase production by using an experimental design. The factors having the strongest influence on chitosanase production were ammonium sulfate concentration, aeration, pH and the interaction between the first two parameters. Optimal conditions for chitosan hydrolysis were pH 5.8 and 54 oC; however, hydrolysis activity drastically decreased at pH 7.0. The enzyme was purified (single-electrophoretic band) by partitioning in an aqueous two-phase system (ATPS), followed by cation-exchange chromatography with a 66% yield. Chitosanase was mainly collected in the top phase (K = 129) of a 22% PEG 1,500, 13% phosphate (pH = 5.8) and 12% NaCl (w/w) solution, and the main protein contaminants were evenly distributed between the phases (K = 1.07). The apparent molecular weight and the isoelectric point of the chitosanase, determined by SDS-PAGE electrophoresis and by isoelectric focalization, were 47 kDa and 8.8, respectively. |
publishDate |
1999 |
dc.date.none.fl_str_mv |
1999-06-01 |
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=S0104-66321999000200011 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0104-66321999000200011 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S0104-66321999000200011 |
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 |
Brazilian Society of Chemical Engineering |
publisher.none.fl_str_mv |
Brazilian Society of Chemical Engineering |
dc.source.none.fl_str_mv |
Brazilian Journal of Chemical Engineering v.16 n.2 1999 reponame:Brazilian Journal of Chemical Engineering instname:Associação Brasileira de Engenharia Química (ABEQ) instacron:ABEQ |
instname_str |
Associação Brasileira de Engenharia Química (ABEQ) |
instacron_str |
ABEQ |
institution |
ABEQ |
reponame_str |
Brazilian Journal of Chemical Engineering |
collection |
Brazilian Journal of Chemical Engineering |
repository.name.fl_str_mv |
Brazilian Journal of Chemical Engineering - Associação Brasileira de Engenharia Química (ABEQ) |
repository.mail.fl_str_mv |
rgiudici@usp.br||rgiudici@usp.br |
_version_ |
1754213170404130816 |