Biochemical characterization and low-resolution SAXS structure of an exo-polygalacturonase from Bacillus licheniformis
Among the structural polymers present in the plant cell wall, pectin is the main component of the middle lamella. This heterogeneous polysaccharide has an α-1,4 galacturonic acid backbone, which can be broken by the enzymatic action of pectinases, such as exo-polygalacturonases, that sequentially cl...
| Autores: | , , , , |
|---|---|
| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2018 |
| País: | Brasil |
| Institución: | Universidade Estadual Paulista (UNESP) |
| Repositorio: | Repositório Institucional da UNESP |
| Idioma: | inglés |
| OAI Identifier: | oai:repositorio.unesp.br:11449/170249 |
| Acceso en línea: | http://dx.doi.org/10.1016/j.nbt.2017.10.001 http://hdl.handle.net/11449/170249 |
| Access Level: | acceso abierto |
| Palabra clave: | Biochemical characterization Exo-polygalacturonase Pectinase SAXS molecular envelope Thermal and pH stability |
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Biochemical characterization and low-resolution SAXS structure of an exo-polygalacturonase from Bacillus licheniformisBiochemical characterizationExo-polygalacturonasePectinaseSAXS molecular envelopeThermal and pH stabilityAmong the structural polymers present in the plant cell wall, pectin is the main component of the middle lamella. This heterogeneous polysaccharide has an α-1,4 galacturonic acid backbone, which can be broken by the enzymatic action of pectinases, such as exo-polygalacturonases, that sequentially cleave pectin from the non-reducing ends, releasing mono or di-galacturonic acid residues. Constant demand for pectinases that better suit industrial requirements has motivated identification and characterization of novel enzymes from diverse sources. Bacillus licheniformis has been used as an important source for bioprospection of several industrial biomolecules, such as surfactants and enzymes, including pectate lyases. Here we cloned, expressed, purified, and biochemically and structurally characterized an exo-polygalacturonase from B. licheniformis (BlExoPG). Its low-resolution molecular envelope was derived from experimental small-angle scattering data (SAXS). Our experimental data revealed that BlExoPG is a monomeric enzyme with optimum pH at 6.5 and optimal temperature of approximately 60 °C, at which it has considerable stability over the broad pH range from 5 to 10. After incubation of the enzyme for 30 min at pH ranging from 5 to 10, no significant loss of the original enzyme activity was observed. Furthermore, the enzyme maintained residual activity of greater than 80% at 50 °C after 15 h of incubation. BlExoPG is more active against polygalacturonic acid as compared to methylated pectin, liberating mono galacturonic acid as a unique product. Its enzymatic parameters are Vmax = 4.18 μM.s−1, Km = 3.25 mgmL−1 and kcat = 2.58 s−1.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Instituto de Física de São Carlos Universidade de São Paulo Avenida Trabalhador Sãocarlense 400Departmento de Física e Biofísica Institudo de Biociências Universidade Estadual Paulista “Júlio de Mesquita Filho” (UNESP)Departmento de Física e Biofísica Institudo de Biociências Universidade Estadual Paulista “Júlio de Mesquita Filho” (UNESP)CNPq: #158752/2015-5FAPESP: #2011/20505-4FAPESP: #2015/13684-0CNPq: #303988/2016-9CNPq: #405191/2015-4CNPq: #440977/2016-9Universidade de São Paulo (USP)Universidade Estadual Paulista (Unesp)2018-12-11T16:49:56Z2018-12-11T16:49:56Z2018-01-25info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article268-274application/pdfhttp://dx.doi.org/10.1016/j.nbt.2017.10.001New Biotechnology, v. 40, p. 268-274.1876-43471871-6784http://hdl.handle.net/11449/17024910.1016/j.nbt.2017.10.0012-s2.0-850307665262-s2.0-85030766526.pdf8213371495151651Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengNew Biotechnology0,967info:eu-repo/semantics/openAccessEvangelista, Danilo Eltonde Araújo, Evandro A.Neto, Mario Oliveira [UNESP]Kadowaki, Marco Antonio SeikiPolikarpov, Igor2023-11-15T06:16:07Zoai:repositorio.unesp.br:11449/170249Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestrepositoriounesp@unesp.bropendoar:29462023-11-15T06:16:07Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
| dc.title.none.fl_str_mv |
Biochemical characterization and low-resolution SAXS structure of an exo-polygalacturonase from Bacillus licheniformis |
| title |
Biochemical characterization and low-resolution SAXS structure of an exo-polygalacturonase from Bacillus licheniformis |
| spellingShingle |
Biochemical characterization and low-resolution SAXS structure of an exo-polygalacturonase from Bacillus licheniformis Evangelista, Danilo Elton Biochemical characterization Exo-polygalacturonase Pectinase SAXS molecular envelope Thermal and pH stability |
| title_short |
Biochemical characterization and low-resolution SAXS structure of an exo-polygalacturonase from Bacillus licheniformis |
| title_full |
Biochemical characterization and low-resolution SAXS structure of an exo-polygalacturonase from Bacillus licheniformis |
| title_fullStr |
Biochemical characterization and low-resolution SAXS structure of an exo-polygalacturonase from Bacillus licheniformis |
| title_full_unstemmed |
Biochemical characterization and low-resolution SAXS structure of an exo-polygalacturonase from Bacillus licheniformis |
| title_sort |
Biochemical characterization and low-resolution SAXS structure of an exo-polygalacturonase from Bacillus licheniformis |
| dc.creator.none.fl_str_mv |
Evangelista, Danilo Elton de Araújo, Evandro A. Neto, Mario Oliveira [UNESP] Kadowaki, Marco Antonio Seiki Polikarpov, Igor |
| author |
Evangelista, Danilo Elton |
| author_facet |
Evangelista, Danilo Elton de Araújo, Evandro A. Neto, Mario Oliveira [UNESP] Kadowaki, Marco Antonio Seiki Polikarpov, Igor |
| author_role |
author |
| author2 |
de Araújo, Evandro A. Neto, Mario Oliveira [UNESP] Kadowaki, Marco Antonio Seiki Polikarpov, Igor |
| author2_role |
author author author author |
| dc.contributor.none.fl_str_mv |
Universidade de São Paulo (USP) Universidade Estadual Paulista (Unesp) |
| dc.subject.por.fl_str_mv |
Biochemical characterization Exo-polygalacturonase Pectinase SAXS molecular envelope Thermal and pH stability |
| topic |
Biochemical characterization Exo-polygalacturonase Pectinase SAXS molecular envelope Thermal and pH stability |
| description |
Among the structural polymers present in the plant cell wall, pectin is the main component of the middle lamella. This heterogeneous polysaccharide has an α-1,4 galacturonic acid backbone, which can be broken by the enzymatic action of pectinases, such as exo-polygalacturonases, that sequentially cleave pectin from the non-reducing ends, releasing mono or di-galacturonic acid residues. Constant demand for pectinases that better suit industrial requirements has motivated identification and characterization of novel enzymes from diverse sources. Bacillus licheniformis has been used as an important source for bioprospection of several industrial biomolecules, such as surfactants and enzymes, including pectate lyases. Here we cloned, expressed, purified, and biochemically and structurally characterized an exo-polygalacturonase from B. licheniformis (BlExoPG). Its low-resolution molecular envelope was derived from experimental small-angle scattering data (SAXS). Our experimental data revealed that BlExoPG is a monomeric enzyme with optimum pH at 6.5 and optimal temperature of approximately 60 °C, at which it has considerable stability over the broad pH range from 5 to 10. After incubation of the enzyme for 30 min at pH ranging from 5 to 10, no significant loss of the original enzyme activity was observed. Furthermore, the enzyme maintained residual activity of greater than 80% at 50 °C after 15 h of incubation. BlExoPG is more active against polygalacturonic acid as compared to methylated pectin, liberating mono galacturonic acid as a unique product. Its enzymatic parameters are Vmax = 4.18 μM.s−1, Km = 3.25 mgmL−1 and kcat = 2.58 s−1. |
| publishDate |
2018 |
| dc.date.none.fl_str_mv |
2018-12-11T16:49:56Z 2018-12-11T16:49:56Z 2018-01-25 |
| 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/10.1016/j.nbt.2017.10.001 New Biotechnology, v. 40, p. 268-274. 1876-4347 1871-6784 http://hdl.handle.net/11449/170249 10.1016/j.nbt.2017.10.001 2-s2.0-85030766526 2-s2.0-85030766526.pdf 8213371495151651 |
| url |
http://dx.doi.org/10.1016/j.nbt.2017.10.001 http://hdl.handle.net/11449/170249 |
| identifier_str_mv |
New Biotechnology, v. 40, p. 268-274. 1876-4347 1871-6784 10.1016/j.nbt.2017.10.001 2-s2.0-85030766526 2-s2.0-85030766526.pdf 8213371495151651 |
| dc.language.iso.fl_str_mv |
eng |
| language |
eng |
| dc.relation.none.fl_str_mv |
New Biotechnology 0,967 |
| dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
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openAccess |
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268-274 application/pdf |
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Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
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Universidade Estadual Paulista (UNESP) |
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UNESP |
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UNESP |
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Repositório Institucional da UNESP |
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Repositório Institucional da UNESP |
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Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
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repositoriounesp@unesp.br |
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