Isolation and characterization of a β-galactosidase from a new Amazon forest strain of Aspergillus niger as a potential accessory enzyme for biomass conversion
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.3109/10242422.2013.801018 http://hdl.handle.net/11449/220044 |
Resumo: | The selection of enzyme-producing fungi is useful to obtain enzymes required to hydrolyze lignocellulosic material and thereby contribute to biomass conversion into fuels and chemicals. Besides cellulases, the presence of accessory enzymes in enzyme cocktails is necessary to enhance hydrolysis efficiency. This study evaluates the production, purification, and biochemical-kinetic characterization of β-galactosidase produced by a new strain of Aspergillus niger (P47C3) isolated from the Amazon Forest. The A. niger (P47C3) was cultured under SmF conditions and β-galactosidase was purified in a three-step purification, using an ultrafiltration membrane, ion exchange (TSK-SP), and gel filtration (Sephacryl S-200). The calculated molecular weight of the purified enzyme was 125 kDa. Optimum pH (4.0) and temperature (55°C) of β-galactosidase activity were determined. The values of the kinetic parameters obtained from p-nitrophenyl-β-D- galactopyranoside (PNPG) hydrolysis were 2.2 mM and 0.285 mM/min for Km and Vmax, respectively. The inhibition of PNPG hydrolysis by β-galactosidase in the presence of the inhibitor galactose gave a Ki value of 5.01 mM. As a precursor to elucidating the tertiary structure using X-ray diffraction, the β-galactosidase was crystallized using 0.2 M Tris-HCl buffer, with 12% PEG 4000 as the precipitation agent; the largest crystals were formed at pH 8.6. These results provide the basis for further structural and functional studies of this accessory enzyme to evaluate its potential biotechnological applications. © 2014 Informa UK, Ltd. |
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Isolation and characterization of a β-galactosidase from a new Amazon forest strain of Aspergillus niger as a potential accessory enzyme for biomass conversionAccessory enzymesAmazonAspergillus nigerBiomassβ-galactosidaseThe selection of enzyme-producing fungi is useful to obtain enzymes required to hydrolyze lignocellulosic material and thereby contribute to biomass conversion into fuels and chemicals. Besides cellulases, the presence of accessory enzymes in enzyme cocktails is necessary to enhance hydrolysis efficiency. This study evaluates the production, purification, and biochemical-kinetic characterization of β-galactosidase produced by a new strain of Aspergillus niger (P47C3) isolated from the Amazon Forest. The A. niger (P47C3) was cultured under SmF conditions and β-galactosidase was purified in a three-step purification, using an ultrafiltration membrane, ion exchange (TSK-SP), and gel filtration (Sephacryl S-200). The calculated molecular weight of the purified enzyme was 125 kDa. Optimum pH (4.0) and temperature (55°C) of β-galactosidase activity were determined. The values of the kinetic parameters obtained from p-nitrophenyl-β-D- galactopyranoside (PNPG) hydrolysis were 2.2 mM and 0.285 mM/min for Km and Vmax, respectively. The inhibition of PNPG hydrolysis by β-galactosidase in the presence of the inhibitor galactose gave a Ki value of 5.01 mM. As a precursor to elucidating the tertiary structure using X-ray diffraction, the β-galactosidase was crystallized using 0.2 M Tris-HCl buffer, with 12% PEG 4000 as the precipitation agent; the largest crystals were formed at pH 8.6. These results provide the basis for further structural and functional studies of this accessory enzyme to evaluate its potential biotechnological applications. © 2014 Informa UK, Ltd.Programa de Pós-graduação em Biotecnologia Universidade Federal de São Carlos (UFSCar), Rod. Washington Luis, Km 235, 13565-905, São Carlos/SPEmbrapa Instrumentação, Rua XV de Novembro, 1452, 13560-970, São Carlos/SPPetersburg Nuclear Physics Institute PNPI RAS Gatchina, Leningrad District 188300Universidade do Estado de São Paulo (USP), Av. Trabalhador São-Carlense 400 Arnold Schimidt, 13566-590, São Carlos/SPUniversidade Federal de São Carlos (UFSCar)Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)PNPI RAS GatchinaUniversidade de São Paulo (USP)Tonelotto, MarianaPirota, Rosangela Donizete Perpetua BuzonDelabona, Priscila Da SilvaBarros, Georgia De Oliveira FigueiredoGolubev, Alexander M.Polikarpov, IgorFarinas, Cristiane Sanchez2022-04-28T18:59:18Z2022-04-28T18:59:18Z2014-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article13-22http://dx.doi.org/10.3109/10242422.2013.801018Biocatalysis and Biotransformation, v. 32, n. 1, p. 13-22, 2014.1029-24461024-2422http://hdl.handle.net/11449/22004410.3109/10242422.2013.8010182-s2.0-84893154287Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengBiocatalysis and Biotransformationinfo:eu-repo/semantics/openAccess2022-04-28T18:59:18Zoai:repositorio.unesp.br:11449/220044Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462022-04-28T18:59:18Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Isolation and characterization of a β-galactosidase from a new Amazon forest strain of Aspergillus niger as a potential accessory enzyme for biomass conversion |
title |
Isolation and characterization of a β-galactosidase from a new Amazon forest strain of Aspergillus niger as a potential accessory enzyme for biomass conversion |
spellingShingle |
Isolation and characterization of a β-galactosidase from a new Amazon forest strain of Aspergillus niger as a potential accessory enzyme for biomass conversion Tonelotto, Mariana Accessory enzymes Amazon Aspergillus niger Biomass β-galactosidase |
title_short |
Isolation and characterization of a β-galactosidase from a new Amazon forest strain of Aspergillus niger as a potential accessory enzyme for biomass conversion |
title_full |
Isolation and characterization of a β-galactosidase from a new Amazon forest strain of Aspergillus niger as a potential accessory enzyme for biomass conversion |
title_fullStr |
Isolation and characterization of a β-galactosidase from a new Amazon forest strain of Aspergillus niger as a potential accessory enzyme for biomass conversion |
title_full_unstemmed |
Isolation and characterization of a β-galactosidase from a new Amazon forest strain of Aspergillus niger as a potential accessory enzyme for biomass conversion |
title_sort |
Isolation and characterization of a β-galactosidase from a new Amazon forest strain of Aspergillus niger as a potential accessory enzyme for biomass conversion |
author |
Tonelotto, Mariana |
author_facet |
Tonelotto, Mariana Pirota, Rosangela Donizete Perpetua Buzon Delabona, Priscila Da Silva Barros, Georgia De Oliveira Figueiredo Golubev, Alexander M. Polikarpov, Igor Farinas, Cristiane Sanchez |
author_role |
author |
author2 |
Pirota, Rosangela Donizete Perpetua Buzon Delabona, Priscila Da Silva Barros, Georgia De Oliveira Figueiredo Golubev, Alexander M. Polikarpov, Igor Farinas, Cristiane Sanchez |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Federal de São Carlos (UFSCar) Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) PNPI RAS Gatchina Universidade de São Paulo (USP) |
dc.contributor.author.fl_str_mv |
Tonelotto, Mariana Pirota, Rosangela Donizete Perpetua Buzon Delabona, Priscila Da Silva Barros, Georgia De Oliveira Figueiredo Golubev, Alexander M. Polikarpov, Igor Farinas, Cristiane Sanchez |
dc.subject.por.fl_str_mv |
Accessory enzymes Amazon Aspergillus niger Biomass β-galactosidase |
topic |
Accessory enzymes Amazon Aspergillus niger Biomass β-galactosidase |
description |
The selection of enzyme-producing fungi is useful to obtain enzymes required to hydrolyze lignocellulosic material and thereby contribute to biomass conversion into fuels and chemicals. Besides cellulases, the presence of accessory enzymes in enzyme cocktails is necessary to enhance hydrolysis efficiency. This study evaluates the production, purification, and biochemical-kinetic characterization of β-galactosidase produced by a new strain of Aspergillus niger (P47C3) isolated from the Amazon Forest. The A. niger (P47C3) was cultured under SmF conditions and β-galactosidase was purified in a three-step purification, using an ultrafiltration membrane, ion exchange (TSK-SP), and gel filtration (Sephacryl S-200). The calculated molecular weight of the purified enzyme was 125 kDa. Optimum pH (4.0) and temperature (55°C) of β-galactosidase activity were determined. The values of the kinetic parameters obtained from p-nitrophenyl-β-D- galactopyranoside (PNPG) hydrolysis were 2.2 mM and 0.285 mM/min for Km and Vmax, respectively. The inhibition of PNPG hydrolysis by β-galactosidase in the presence of the inhibitor galactose gave a Ki value of 5.01 mM. As a precursor to elucidating the tertiary structure using X-ray diffraction, the β-galactosidase was crystallized using 0.2 M Tris-HCl buffer, with 12% PEG 4000 as the precipitation agent; the largest crystals were formed at pH 8.6. These results provide the basis for further structural and functional studies of this accessory enzyme to evaluate its potential biotechnological applications. © 2014 Informa UK, Ltd. |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014-01-01 2022-04-28T18:59:18Z 2022-04-28T18:59:18Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.3109/10242422.2013.801018 Biocatalysis and Biotransformation, v. 32, n. 1, p. 13-22, 2014. 1029-2446 1024-2422 http://hdl.handle.net/11449/220044 10.3109/10242422.2013.801018 2-s2.0-84893154287 |
url |
http://dx.doi.org/10.3109/10242422.2013.801018 http://hdl.handle.net/11449/220044 |
identifier_str_mv |
Biocatalysis and Biotransformation, v. 32, n. 1, p. 13-22, 2014. 1029-2446 1024-2422 10.3109/10242422.2013.801018 2-s2.0-84893154287 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Biocatalysis and Biotransformation |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
13-22 |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
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1803047461106745344 |