Bg10: A novel metagenomics alcohol-tolerant and glucose-stimulated gh1 β-glucosidase suitable for lactose-free milk preparation
Autor(a) principal: | |
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Data de Publicação: | 2016 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1371/journal.pone.0167932 http://hdl.handle.net/11449/173999 |
Resumo: | New β-glucosidases with product (glucose) or ethanol tolerances are greatly desired to make industrial processes more marketable and efficient. Therefore, this report describes the in silico/vitro characterization of Bg10, a metagenomically derived homodimeric β-glucosidase that exhibited a Vmax of 10.81 ± 0.43 μM min-1 , Kcat of 175.1± 6.91 min-1 , and Km of 0.49 ± 0.12 mM at a neutral pH and 37°C when pNP-β-D-glucopyranoside was used as the substrate, and the enzyme retained greater than 80% activity within the respective pH and temperature ranges of 6.5 to 8.0 and 35 to 40°C. The enzyme was stimulated by its product, glucose; consequently, the Bg10 activity against 50 and 100 mM of glucose were increased by 36.8% and 22%, respectively, while half of the activity was retained at 350 mM. Moreover, the Bg10 was able to hydrolyse 55% (milk sample) and 100% (purified sugar) of the lactose at low (6°C) and optimum (37°C) temperatures, respectively, suggesting the possibility of further optimization of the reaction for lactose-free dairy production. In addition, the enzyme was able to fully hydrolyse 40 mM of cellobiose at one hour and was tolerant to ethanol up to concentrations of 500 mM (86% of activity), while a 1 M concentration still resulted in 41% residual activity, which could be interesting for biofuel production. |
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spelling |
Bg10: A novel metagenomics alcohol-tolerant and glucose-stimulated gh1 β-glucosidase suitable for lactose-free milk preparationNew β-glucosidases with product (glucose) or ethanol tolerances are greatly desired to make industrial processes more marketable and efficient. Therefore, this report describes the in silico/vitro characterization of Bg10, a metagenomically derived homodimeric β-glucosidase that exhibited a Vmax of 10.81 ± 0.43 μM min-1 , Kcat of 175.1± 6.91 min-1 , and Km of 0.49 ± 0.12 mM at a neutral pH and 37°C when pNP-β-D-glucopyranoside was used as the substrate, and the enzyme retained greater than 80% activity within the respective pH and temperature ranges of 6.5 to 8.0 and 35 to 40°C. The enzyme was stimulated by its product, glucose; consequently, the Bg10 activity against 50 and 100 mM of glucose were increased by 36.8% and 22%, respectively, while half of the activity was retained at 350 mM. Moreover, the Bg10 was able to hydrolyse 55% (milk sample) and 100% (purified sugar) of the lactose at low (6°C) and optimum (37°C) temperatures, respectively, suggesting the possibility of further optimization of the reaction for lactose-free dairy production. In addition, the enzyme was able to fully hydrolyse 40 mM of cellobiose at one hour and was tolerant to ethanol up to concentrations of 500 mM (86% of activity), while a 1 M concentration still resulted in 41% residual activity, which could be interesting for biofuel production.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Department of Technology São Paulo State University (Unesp) School of Agricultural and Veterinarian Sciences Jaboticabal, Via de Acesso Prof. Paulo Donato Castellane S/NMolecular Biology Laboratory Institute for Research in Bioenergy (IPBEN) UNESPAgricultural Microbiology Postgraduate Program of UNESPDepartment of Technology São Paulo State University (Unesp) School of Agricultural and Veterinarian Sciences Jaboticabal, Via de Acesso Prof. Paulo Donato Castellane S/NMolecular Biology Laboratory Institute for Research in Bioenergy (IPBEN) UNESPAgricultural Microbiology Postgraduate Program of UNESPFAPESP: ESGP 2011/10981-3Universidade Estadual Paulista (Unesp)Gomes-Pepe, Elisângela Soares [UNESP]Sierra, Elwi Guillermo Machado [UNESP]Pereira, Mariana Rangel [UNESP]Castellane, Tereza Cristina Luque [UNESP]De Lemos, Eliana Gertrudes Macedo [UNESP]2018-12-11T17:08:41Z2018-12-11T17:08:41Z2016-12-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://dx.doi.org/10.1371/journal.pone.0167932PLoS ONE, v. 11, n. 12, 2016.1932-6203http://hdl.handle.net/11449/17399910.1371/journal.pone.01679322-s2.0-850074223352-s2.0-85007422335.pdfScopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengPLoS ONE1,164info:eu-repo/semantics/openAccess2024-06-07T15:31:35Zoai:repositorio.unesp.br:11449/173999Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-06-07T15:31:35Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Bg10: A novel metagenomics alcohol-tolerant and glucose-stimulated gh1 β-glucosidase suitable for lactose-free milk preparation |
title |
Bg10: A novel metagenomics alcohol-tolerant and glucose-stimulated gh1 β-glucosidase suitable for lactose-free milk preparation |
spellingShingle |
Bg10: A novel metagenomics alcohol-tolerant and glucose-stimulated gh1 β-glucosidase suitable for lactose-free milk preparation Gomes-Pepe, Elisângela Soares [UNESP] |
title_short |
Bg10: A novel metagenomics alcohol-tolerant and glucose-stimulated gh1 β-glucosidase suitable for lactose-free milk preparation |
title_full |
Bg10: A novel metagenomics alcohol-tolerant and glucose-stimulated gh1 β-glucosidase suitable for lactose-free milk preparation |
title_fullStr |
Bg10: A novel metagenomics alcohol-tolerant and glucose-stimulated gh1 β-glucosidase suitable for lactose-free milk preparation |
title_full_unstemmed |
Bg10: A novel metagenomics alcohol-tolerant and glucose-stimulated gh1 β-glucosidase suitable for lactose-free milk preparation |
title_sort |
Bg10: A novel metagenomics alcohol-tolerant and glucose-stimulated gh1 β-glucosidase suitable for lactose-free milk preparation |
author |
Gomes-Pepe, Elisângela Soares [UNESP] |
author_facet |
Gomes-Pepe, Elisângela Soares [UNESP] Sierra, Elwi Guillermo Machado [UNESP] Pereira, Mariana Rangel [UNESP] Castellane, Tereza Cristina Luque [UNESP] De Lemos, Eliana Gertrudes Macedo [UNESP] |
author_role |
author |
author2 |
Sierra, Elwi Guillermo Machado [UNESP] Pereira, Mariana Rangel [UNESP] Castellane, Tereza Cristina Luque [UNESP] De Lemos, Eliana Gertrudes Macedo [UNESP] |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Gomes-Pepe, Elisângela Soares [UNESP] Sierra, Elwi Guillermo Machado [UNESP] Pereira, Mariana Rangel [UNESP] Castellane, Tereza Cristina Luque [UNESP] De Lemos, Eliana Gertrudes Macedo [UNESP] |
description |
New β-glucosidases with product (glucose) or ethanol tolerances are greatly desired to make industrial processes more marketable and efficient. Therefore, this report describes the in silico/vitro characterization of Bg10, a metagenomically derived homodimeric β-glucosidase that exhibited a Vmax of 10.81 ± 0.43 μM min-1 , Kcat of 175.1± 6.91 min-1 , and Km of 0.49 ± 0.12 mM at a neutral pH and 37°C when pNP-β-D-glucopyranoside was used as the substrate, and the enzyme retained greater than 80% activity within the respective pH and temperature ranges of 6.5 to 8.0 and 35 to 40°C. The enzyme was stimulated by its product, glucose; consequently, the Bg10 activity against 50 and 100 mM of glucose were increased by 36.8% and 22%, respectively, while half of the activity was retained at 350 mM. Moreover, the Bg10 was able to hydrolyse 55% (milk sample) and 100% (purified sugar) of the lactose at low (6°C) and optimum (37°C) temperatures, respectively, suggesting the possibility of further optimization of the reaction for lactose-free dairy production. In addition, the enzyme was able to fully hydrolyse 40 mM of cellobiose at one hour and was tolerant to ethanol up to concentrations of 500 mM (86% of activity), while a 1 M concentration still resulted in 41% residual activity, which could be interesting for biofuel production. |
publishDate |
2016 |
dc.date.none.fl_str_mv |
2016-12-01 2018-12-11T17:08:41Z 2018-12-11T17:08:41Z |
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/10.1371/journal.pone.0167932 PLoS ONE, v. 11, n. 12, 2016. 1932-6203 http://hdl.handle.net/11449/173999 10.1371/journal.pone.0167932 2-s2.0-85007422335 2-s2.0-85007422335.pdf |
url |
http://dx.doi.org/10.1371/journal.pone.0167932 http://hdl.handle.net/11449/173999 |
identifier_str_mv |
PLoS ONE, v. 11, n. 12, 2016. 1932-6203 10.1371/journal.pone.0167932 2-s2.0-85007422335 2-s2.0-85007422335.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
PLoS ONE 1,164 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
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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