Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34

Autores
Hashim, Suhaila O.; Delgado, Osvaldo Daniel; Martinez, Maria Alejandra; Kaul, Rajni-Hatti; Mulaa, Francis J.; Mattiasson, Bo
Año de publicación
2005
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The gene encoding Amy 34, a maltohexaose-forming -amylase from Bacillus halodurans LBK 34 isolated from Lake Bogoria, Kenya, was cloned and sequenced. The mature peptide consists of 958 amino acids with a theoretical molecular weight of 107.2 kDa and pI 4.41, respectively. The gene was expressed in Escherichia coli and the recombinant enzyme purified to homogeneity by a combination of metal chelate affinity and size exclusion chromatography. The pure enzyme exhibited optimum activity at 60 ◦C and pH 10.5–11.5. The enzyme retained over 60% activity after incubation at 55 ◦C for 4 h and was most stable at pH 9.0. Complete inhibition of enzyme activity was observed in presence of 5 mM Cu2+, Fe2+, Fe3+, Mn2+ and 5 mM EDTA. The enzyme displayed 80% of its original activity in presence of 1% (w/v) SDS and was stable in presence of up to 5 mM DTT. Maltohexaose (G6) was the main initial product of starch hydrolysis while other products formed were G4 > G2 > G5 > G3 and G1. The main end product of the enzyme’s action on amylose, amylopectin and maltodextrin is maltotetraose. Amy 34 could not hydrolyse pullulan, and -cyclodextrin but could hydrolyse -cyclodextrin to produce glucose, maltose and maltotetraose. Maltotetraose was the smallest -(1–4) linked maltooligosaccharide that could be hydrolysed by the enzyme
Fil: Hashim, Suhaila O.. Lund University; Suecia
Fil: Delgado, Osvaldo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina
Fil: Martinez, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina
Fil: Kaul, Rajni-Hatti. Lund University; Suecia
Fil: Mulaa, Francis J.. University of Nairobi; Kenia
Fil: Mattiasson, Bo. Lund University; Suecia
Materia
Alkaliphile
Amylase
B. Halodurans
Maltohexaose
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/41857
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/41857
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34Hashim, Suhaila O.Delgado, Osvaldo DanielMartinez, Maria AlejandraKaul, Rajni-HattiMulaa, Francis J.Mattiasson, BoAlkaliphileAmylaseB. HaloduransMaltohexaosehttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The gene encoding Amy 34, a maltohexaose-forming -amylase from Bacillus halodurans LBK 34 isolated from Lake Bogoria, Kenya, was cloned and sequenced. The mature peptide consists of 958 amino acids with a theoretical molecular weight of 107.2 kDa and pI 4.41, respectively. The gene was expressed in Escherichia coli and the recombinant enzyme purified to homogeneity by a combination of metal chelate affinity and size exclusion chromatography. The pure enzyme exhibited optimum activity at 60 ◦C and pH 10.5–11.5. The enzyme retained over 60% activity after incubation at 55 ◦C for 4 h and was most stable at pH 9.0. Complete inhibition of enzyme activity was observed in presence of 5 mM Cu2+, Fe2+, Fe3+, Mn2+ and 5 mM EDTA. The enzyme displayed 80% of its original activity in presence of 1% (w/v) SDS and was stable in presence of up to 5 mM DTT. Maltohexaose (G6) was the main initial product of starch hydrolysis while other products formed were G4 > G2 > G5 > G3 and G1. The main end product of the enzyme’s action on amylose, amylopectin and maltodextrin is maltotetraose. Amy 34 could not hydrolyse pullulan, and -cyclodextrin but could hydrolyse -cyclodextrin to produce glucose, maltose and maltotetraose. Maltotetraose was the smallest -(1–4) linked maltooligosaccharide that could be hydrolysed by the enzymeFil: Hashim, Suhaila O.. Lund University; SueciaFil: Delgado, Osvaldo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Martinez, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; ArgentinaFil: Kaul, Rajni-Hatti. Lund University; SueciaFil: Mulaa, Francis J.. University of Nairobi; KeniaFil: Mattiasson, Bo. Lund University; SueciaElsevier Science Inc2005-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/41857Hashim, Suhaila O.; Delgado, Osvaldo Daniel; Martinez, Maria Alejandra; Kaul, Rajni-Hatti; Mulaa, Francis J.; et al.; Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34; Elsevier Science Inc; Enzyme and Microbial Technology; 36; 1; 12-2005; 139-1460141-0229CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.enzmictec.2004.07.017info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0141022904002947info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:57:51Zoai:ri.conicet.gov.ar:11336/41857instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 09:57:51.363CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34
title Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34
spellingShingle Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34
Hashim, Suhaila O.
Alkaliphile
Amylase
B. Halodurans
Maltohexaose
title_short Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34
title_full Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34
title_fullStr Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34
title_full_unstemmed Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34
title_sort Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34
dc.creator.none.fl_str_mv Hashim, Suhaila O.
Delgado, Osvaldo Daniel
Martinez, Maria Alejandra
Kaul, Rajni-Hatti
Mulaa, Francis J.
Mattiasson, Bo
author Hashim, Suhaila O.
author_facet Hashim, Suhaila O.
Delgado, Osvaldo Daniel
Martinez, Maria Alejandra
Kaul, Rajni-Hatti
Mulaa, Francis J.
Mattiasson, Bo
author_role author
author2 Delgado, Osvaldo Daniel
Martinez, Maria Alejandra
Kaul, Rajni-Hatti
Mulaa, Francis J.
Mattiasson, Bo
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Alkaliphile
Amylase
B. Halodurans
Maltohexaose
topic Alkaliphile
Amylase
B. Halodurans
Maltohexaose
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The gene encoding Amy 34, a maltohexaose-forming -amylase from Bacillus halodurans LBK 34 isolated from Lake Bogoria, Kenya, was cloned and sequenced. The mature peptide consists of 958 amino acids with a theoretical molecular weight of 107.2 kDa and pI 4.41, respectively. The gene was expressed in Escherichia coli and the recombinant enzyme purified to homogeneity by a combination of metal chelate affinity and size exclusion chromatography. The pure enzyme exhibited optimum activity at 60 ◦C and pH 10.5–11.5. The enzyme retained over 60% activity after incubation at 55 ◦C for 4 h and was most stable at pH 9.0. Complete inhibition of enzyme activity was observed in presence of 5 mM Cu2+, Fe2+, Fe3+, Mn2+ and 5 mM EDTA. The enzyme displayed 80% of its original activity in presence of 1% (w/v) SDS and was stable in presence of up to 5 mM DTT. Maltohexaose (G6) was the main initial product of starch hydrolysis while other products formed were G4 > G2 > G5 > G3 and G1. The main end product of the enzyme’s action on amylose, amylopectin and maltodextrin is maltotetraose. Amy 34 could not hydrolyse pullulan, and -cyclodextrin but could hydrolyse -cyclodextrin to produce glucose, maltose and maltotetraose. Maltotetraose was the smallest -(1–4) linked maltooligosaccharide that could be hydrolysed by the enzyme
Fil: Hashim, Suhaila O.. Lund University; Suecia
Fil: Delgado, Osvaldo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina
Fil: Martinez, Maria Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Planta Piloto de Procesos Industriales Microbiológicos; Argentina
Fil: Kaul, Rajni-Hatti. Lund University; Suecia
Fil: Mulaa, Francis J.. University of Nairobi; Kenia
Fil: Mattiasson, Bo. Lund University; Suecia
description The gene encoding Amy 34, a maltohexaose-forming -amylase from Bacillus halodurans LBK 34 isolated from Lake Bogoria, Kenya, was cloned and sequenced. The mature peptide consists of 958 amino acids with a theoretical molecular weight of 107.2 kDa and pI 4.41, respectively. The gene was expressed in Escherichia coli and the recombinant enzyme purified to homogeneity by a combination of metal chelate affinity and size exclusion chromatography. The pure enzyme exhibited optimum activity at 60 ◦C and pH 10.5–11.5. The enzyme retained over 60% activity after incubation at 55 ◦C for 4 h and was most stable at pH 9.0. Complete inhibition of enzyme activity was observed in presence of 5 mM Cu2+, Fe2+, Fe3+, Mn2+ and 5 mM EDTA. The enzyme displayed 80% of its original activity in presence of 1% (w/v) SDS and was stable in presence of up to 5 mM DTT. Maltohexaose (G6) was the main initial product of starch hydrolysis while other products formed were G4 > G2 > G5 > G3 and G1. The main end product of the enzyme’s action on amylose, amylopectin and maltodextrin is maltotetraose. Amy 34 could not hydrolyse pullulan, and -cyclodextrin but could hydrolyse -cyclodextrin to produce glucose, maltose and maltotetraose. Maltotetraose was the smallest -(1–4) linked maltooligosaccharide that could be hydrolysed by the enzyme
publishDate 2005
dc.date.none.fl_str_mv 2005-12
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/41857
Hashim, Suhaila O.; Delgado, Osvaldo Daniel; Martinez, Maria Alejandra; Kaul, Rajni-Hatti; Mulaa, Francis J.; et al.; Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34; Elsevier Science Inc; Enzyme and Microbial Technology; 36; 1; 12-2005; 139-146
0141-0229
CONICET Digital
CONICET
url http://hdl.handle.net/11336/41857
identifier_str_mv Hashim, Suhaila O.; Delgado, Osvaldo Daniel; Martinez, Maria Alejandra; Kaul, Rajni-Hatti; Mulaa, Francis J.; et al.; Alkaline active maltohexaose-forming α-amylase from Bacillus halodurans LBK 34; Elsevier Science Inc; Enzyme and Microbial Technology; 36; 1; 12-2005; 139-146
0141-0229
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.enzmictec.2004.07.017
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0141022904002947
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science Inc
publisher.none.fl_str_mv Elsevier Science Inc
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1842269488369631232
score 13.13397

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