Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning

Detalhes bibliográficos
Autor(a) principal: Silva,Maria Estela da
Data de Publicação: 1999
Outros Autores: Franco,Telma Teixeira
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Revista de Microbiologia
Texto Completo: http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37141999000400006
Resumo: This work investigated the partitioning of b-galactosidase from Kluyveromyces fragilis in aqueous two-phase systems (ATPS) by bioaffinity. PEG 4000 was chemically activated with thresyl chloride, and the biospecific ligand p-aminophenyl 1-thio-b-D-galactopyranoside (APGP) was attached to the activated PEG 4000. A new two-step method for extraction and purification of the enzyme b-galactosidase from Kluyveromyces fragilis was developed. In the first step, a system composed of 6% PEG 4000-APGP and 8% dextran 505 was used, where b-galactosidase was strongly partitioned to the top phase (K = 2,330). In the second step, a system formed of 13% PEG-APGP and 9% phosphate salt was used to revert the value of the partition coefficient of b-galactosidase (K = 2 x 10-5) in order to provide the purification and recovery of 39% of the enzyme in the bottom salt-rich phase.
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spelling Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioningb-galactosidaseaqueous two-phase systemsprotein purificationdownstream-processingaffinityThis work investigated the partitioning of b-galactosidase from Kluyveromyces fragilis in aqueous two-phase systems (ATPS) by bioaffinity. PEG 4000 was chemically activated with thresyl chloride, and the biospecific ligand p-aminophenyl 1-thio-b-D-galactopyranoside (APGP) was attached to the activated PEG 4000. A new two-step method for extraction and purification of the enzyme b-galactosidase from Kluyveromyces fragilis was developed. In the first step, a system composed of 6% PEG 4000-APGP and 8% dextran 505 was used, where b-galactosidase was strongly partitioned to the top phase (K = 2,330). In the second step, a system formed of 13% PEG-APGP and 9% phosphate salt was used to revert the value of the partition coefficient of b-galactosidase (K = 2 x 10-5) in order to provide the purification and recovery of 39% of the enzyme in the bottom salt-rich phase.Sociedade Brasileira de Microbiologia1999-12-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37141999000400006Revista de Microbiologia v.30 n.4 1999reponame:Revista de Microbiologiainstname:Sociedade Brasileira de Microbiologia (SBM)instacron:SBM10.1590/S0001-37141999000400006info:eu-repo/semantics/openAccessSilva,Maria Estela daFranco,Telma Teixeiraeng2000-04-20T00:00:00Zoai:scielo:S0001-37141999000400006Revistahttps://www.scielo.br/j/rm/ONGhttps://old.scielo.br/oai/scielo-oai.phpbjm@sbmicrobiologia.org.br||revmicro@icb.usp.br0001-37140001-3714opendoar:2000-04-20T00:00Revista de Microbiologia - Sociedade Brasileira de Microbiologia (SBM)false
dc.title.none.fl_str_mv Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning
title Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning
spellingShingle Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning
Silva,Maria Estela da
b-galactosidase
aqueous two-phase systems
protein purification
downstream-processing
affinity
title_short Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning
title_full Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning
title_fullStr Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning
title_full_unstemmed Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning
title_sort Purification of microbial b-galactosidase from Kluyveromyces fragilis by bioaffinity partitioning
author Silva,Maria Estela da
author_facet Silva,Maria Estela da
Franco,Telma Teixeira
author_role author
author2 Franco,Telma Teixeira
author2_role author
dc.contributor.author.fl_str_mv Silva,Maria Estela da
Franco,Telma Teixeira
dc.subject.por.fl_str_mv b-galactosidase
aqueous two-phase systems
protein purification
downstream-processing
affinity
topic b-galactosidase
aqueous two-phase systems
protein purification
downstream-processing
affinity
description This work investigated the partitioning of b-galactosidase from Kluyveromyces fragilis in aqueous two-phase systems (ATPS) by bioaffinity. PEG 4000 was chemically activated with thresyl chloride, and the biospecific ligand p-aminophenyl 1-thio-b-D-galactopyranoside (APGP) was attached to the activated PEG 4000. A new two-step method for extraction and purification of the enzyme b-galactosidase from Kluyveromyces fragilis was developed. In the first step, a system composed of 6% PEG 4000-APGP and 8% dextran 505 was used, where b-galactosidase was strongly partitioned to the top phase (K = 2,330). In the second step, a system formed of 13% PEG-APGP and 9% phosphate salt was used to revert the value of the partition coefficient of b-galactosidase (K = 2 x 10-5) in order to provide the purification and recovery of 39% of the enzyme in the bottom salt-rich phase.
publishDate 1999
dc.date.none.fl_str_mv 1999-12-01
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=S0001-37141999000400006
url http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0001-37141999000400006
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 10.1590/S0001-37141999000400006
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 Revista de Microbiologia v.30 n.4 1999
reponame:Revista de Microbiologia
instname:Sociedade Brasileira de Microbiologia (SBM)
instacron:SBM
instname_str Sociedade Brasileira de Microbiologia (SBM)
instacron_str SBM
institution SBM
reponame_str Revista de Microbiologia
collection Revista de Microbiologia
repository.name.fl_str_mv Revista de Microbiologia - Sociedade Brasileira de Microbiologia (SBM)
repository.mail.fl_str_mv bjm@sbmicrobiologia.org.br||revmicro@icb.usp.br
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