Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradans

Autores
Wayllace, Natael Maximiliano; Hedin, Nicolas; Busi, María Victoria; Gomez Casati, Diego Fabian
Año de publicación
2021
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Glucoamylases (GAs) are hydrolytic enzymes also known as amyloglucosidases, glucan 1,4-alphaglucosidases or exo-1,4-1,6 bonds) from the non- -Dglucose. These are typically microbial enzymes present in archaea, bacteria and fungi but absent in animals and plants, and they are classified into the GH15 family of glycoside hydrolases (www.cazy.org).-amylases and pullulanases) occurs in the process of saccharification of partially processed starch or dextrins to obtain glucose. Currently, there is strong interest in finding GAs with a better performance at low temperatures because these enzymes would avoid the heating requirement in some industrial processes such as starch saccharification among others, and, in this way, production costs could be minimized. Saccharophagus degradans is a  gramnegative marine bacterium. It is the most versatile bacterium in terms of the degradation of complex polymers (CP) found to date. It is capable to degrade at least 10 complex polymers such as starch, agar, laminarin, cellulose, pectin, alginate, chitin, fucoidan, pectin, pullulan, and xylan at high rate. The objective of this work is to carry out the structural characterization and functional properties of SdGA, a novel glucoamylase (GA) from S. degradans. The enzyme is composed mainly of a N-terminal GH15_N domainlinked to a C-terminal catalytic domain (CD) found in the GH15 family of glycosylhydrolases with an overall structure similar to other bacterial GAs. The protein was successfully expressed in Escherichia coli cells, purified and its biochemical properties were investigated. SdGA showed maximum activity at 39°C and pH 6.0. The enzyme has high activity in a wide range, from low to mild temperatures, like cold-adapted enzymes. It showed the same maximum activity in the range of 0 1.0 M NaCl like salt-tolerant amylases.By thermal inactivation assays, we determined that SdGA is thermolabile at temperatures above 42°C and we found that glycerol 10% (V/V), acarbose 0.1 mM and NaCl 1 M stabilized the enzyme. Furthermore, we analyze the CD of SdGA, other cold-adapted, psychrophilic and thermostable GAs and we found that SdGA has a larger CD due to various amino acid insertions and a higher content of flexible residues compared to other thermostable GAs. These characteristics of SdGA allow it to be classified as a coldadaptedenzyme but also, a salt-tolerant enzyme. We propose that this novel SdGA, might have potential applications for use in different industrial processes that require an efficient alpha glucosidase activity at low/mild temperatures, such as biofuel production.
Fil: Wayllace, Natael Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina
Fil: Hedin, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina
Fil: Busi, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina
Fil: Gomez Casati, Diego Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina
Tercer Encuentro de Red Argentina de Tecnología Enzimática; Primer Workshop de la Red Argentina de Tecnología Enzimática
Rosario
Argentina
Red Argentina de Tecnología Enzimática
Materia
ENZIMAS
BACTERIAS
POLISACÁRIDOS
GLUCOAMILASA
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:11336/204462
Acceder
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network_name_str CONICET Digital (CONICET)
spelling Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradansWayllace, Natael MaximilianoHedin, NicolasBusi, María VictoriaGomez Casati, Diego FabianENZIMASBACTERIASPOLISACÁRIDOSGLUCOAMILASAhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Glucoamylases (GAs) are hydrolytic enzymes also known as amyloglucosidases, glucan 1,4-alphaglucosidases or exo-1,4-1,6 bonds) from the non- -Dglucose. These are typically microbial enzymes present in archaea, bacteria and fungi but absent in animals and plants, and they are classified into the GH15 family of glycoside hydrolases (www.cazy.org).-amylases and pullulanases) occurs in the process of saccharification of partially processed starch or dextrins to obtain glucose. Currently, there is strong interest in finding GAs with a better performance at low temperatures because these enzymes would avoid the heating requirement in some industrial processes such as starch saccharification among others, and, in this way, production costs could be minimized. Saccharophagus degradans is a  gramnegative marine bacterium. It is the most versatile bacterium in terms of the degradation of complex polymers (CP) found to date. It is capable to degrade at least 10 complex polymers such as starch, agar, laminarin, cellulose, pectin, alginate, chitin, fucoidan, pectin, pullulan, and xylan at high rate. The objective of this work is to carry out the structural characterization and functional properties of SdGA, a novel glucoamylase (GA) from S. degradans. The enzyme is composed mainly of a N-terminal GH15_N domainlinked to a C-terminal catalytic domain (CD) found in the GH15 family of glycosylhydrolases with an overall structure similar to other bacterial GAs. The protein was successfully expressed in Escherichia coli cells, purified and its biochemical properties were investigated. SdGA showed maximum activity at 39°C and pH 6.0. The enzyme has high activity in a wide range, from low to mild temperatures, like cold-adapted enzymes. It showed the same maximum activity in the range of 0 1.0 M NaCl like salt-tolerant amylases.By thermal inactivation assays, we determined that SdGA is thermolabile at temperatures above 42°C and we found that glycerol 10% (V/V), acarbose 0.1 mM and NaCl 1 M stabilized the enzyme. Furthermore, we analyze the CD of SdGA, other cold-adapted, psychrophilic and thermostable GAs and we found that SdGA has a larger CD due to various amino acid insertions and a higher content of flexible residues compared to other thermostable GAs. These characteristics of SdGA allow it to be classified as a coldadaptedenzyme but also, a salt-tolerant enzyme. We propose that this novel SdGA, might have potential applications for use in different industrial processes that require an efficient alpha glucosidase activity at low/mild temperatures, such as biofuel production.Fil: Wayllace, Natael Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaFil: Hedin, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaFil: Busi, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaFil: Gomez Casati, Diego Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; ArgentinaTercer Encuentro de Red Argentina de Tecnología Enzimática; Primer Workshop de la Red Argentina de Tecnología EnzimáticaRosarioArgentinaRed Argentina de Tecnología EnzimáticaRed Argentina de Tecnología Enzimática2021info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectEncuentroBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/204462Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradans; Tercer Encuentro de Red Argentina de Tecnología Enzimática; Primer Workshop de la Red Argentina de Tecnología Enzimática; Rosario; Argentina; 2021; 61-62CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.redtez.com.ar/wp-content/uploads/WorkshopRedTEz2021_BookAbstracts.pdfInternacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:23:47Zoai:ri.conicet.gov.ar:11336/204462instacron: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:34982025-09-10 13:23:47.393CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradans
title Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradans
spellingShingle Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradans
Wayllace, Natael Maximiliano
ENZIMAS
BACTERIAS
POLISACÁRIDOS
GLUCOAMILASA
title_short Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradans
title_full Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradans
title_fullStr Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradans
title_full_unstemmed Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradans
title_sort Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradans
dc.creator.none.fl_str_mv Wayllace, Natael Maximiliano
Hedin, Nicolas
Busi, María Victoria
Gomez Casati, Diego Fabian
author Wayllace, Natael Maximiliano
author_facet Wayllace, Natael Maximiliano
Hedin, Nicolas
Busi, María Victoria
Gomez Casati, Diego Fabian
author_role author
author2 Hedin, Nicolas
Busi, María Victoria
Gomez Casati, Diego Fabian
author2_role author
author
author
dc.subject.none.fl_str_mv ENZIMAS
BACTERIAS
POLISACÁRIDOS
GLUCOAMILASA
topic ENZIMAS
BACTERIAS
POLISACÁRIDOS
GLUCOAMILASA
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Glucoamylases (GAs) are hydrolytic enzymes also known as amyloglucosidases, glucan 1,4-alphaglucosidases or exo-1,4-1,6 bonds) from the non- -Dglucose. These are typically microbial enzymes present in archaea, bacteria and fungi but absent in animals and plants, and they are classified into the GH15 family of glycoside hydrolases (www.cazy.org).-amylases and pullulanases) occurs in the process of saccharification of partially processed starch or dextrins to obtain glucose. Currently, there is strong interest in finding GAs with a better performance at low temperatures because these enzymes would avoid the heating requirement in some industrial processes such as starch saccharification among others, and, in this way, production costs could be minimized. Saccharophagus degradans is a  gramnegative marine bacterium. It is the most versatile bacterium in terms of the degradation of complex polymers (CP) found to date. It is capable to degrade at least 10 complex polymers such as starch, agar, laminarin, cellulose, pectin, alginate, chitin, fucoidan, pectin, pullulan, and xylan at high rate. The objective of this work is to carry out the structural characterization and functional properties of SdGA, a novel glucoamylase (GA) from S. degradans. The enzyme is composed mainly of a N-terminal GH15_N domainlinked to a C-terminal catalytic domain (CD) found in the GH15 family of glycosylhydrolases with an overall structure similar to other bacterial GAs. The protein was successfully expressed in Escherichia coli cells, purified and its biochemical properties were investigated. SdGA showed maximum activity at 39°C and pH 6.0. The enzyme has high activity in a wide range, from low to mild temperatures, like cold-adapted enzymes. It showed the same maximum activity in the range of 0 1.0 M NaCl like salt-tolerant amylases.By thermal inactivation assays, we determined that SdGA is thermolabile at temperatures above 42°C and we found that glycerol 10% (V/V), acarbose 0.1 mM and NaCl 1 M stabilized the enzyme. Furthermore, we analyze the CD of SdGA, other cold-adapted, psychrophilic and thermostable GAs and we found that SdGA has a larger CD due to various amino acid insertions and a higher content of flexible residues compared to other thermostable GAs. These characteristics of SdGA allow it to be classified as a coldadaptedenzyme but also, a salt-tolerant enzyme. We propose that this novel SdGA, might have potential applications for use in different industrial processes that require an efficient alpha glucosidase activity at low/mild temperatures, such as biofuel production.
Fil: Wayllace, Natael Maximiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina
Fil: Hedin, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina
Fil: Busi, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina
Fil: Gomez Casati, Diego Fabian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Centro de Estudios Fotosintéticos y Bioquímicos. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Centro de Estudios Fotosintéticos y Bioquímicos; Argentina
Tercer Encuentro de Red Argentina de Tecnología Enzimática; Primer Workshop de la Red Argentina de Tecnología Enzimática
Rosario
Argentina
Red Argentina de Tecnología Enzimática
description Glucoamylases (GAs) are hydrolytic enzymes also known as amyloglucosidases, glucan 1,4-alphaglucosidases or exo-1,4-1,6 bonds) from the non- -Dglucose. These are typically microbial enzymes present in archaea, bacteria and fungi but absent in animals and plants, and they are classified into the GH15 family of glycoside hydrolases (www.cazy.org).-amylases and pullulanases) occurs in the process of saccharification of partially processed starch or dextrins to obtain glucose. Currently, there is strong interest in finding GAs with a better performance at low temperatures because these enzymes would avoid the heating requirement in some industrial processes such as starch saccharification among others, and, in this way, production costs could be minimized. Saccharophagus degradans is a  gramnegative marine bacterium. It is the most versatile bacterium in terms of the degradation of complex polymers (CP) found to date. It is capable to degrade at least 10 complex polymers such as starch, agar, laminarin, cellulose, pectin, alginate, chitin, fucoidan, pectin, pullulan, and xylan at high rate. The objective of this work is to carry out the structural characterization and functional properties of SdGA, a novel glucoamylase (GA) from S. degradans. The enzyme is composed mainly of a N-terminal GH15_N domainlinked to a C-terminal catalytic domain (CD) found in the GH15 family of glycosylhydrolases with an overall structure similar to other bacterial GAs. The protein was successfully expressed in Escherichia coli cells, purified and its biochemical properties were investigated. SdGA showed maximum activity at 39°C and pH 6.0. The enzyme has high activity in a wide range, from low to mild temperatures, like cold-adapted enzymes. It showed the same maximum activity in the range of 0 1.0 M NaCl like salt-tolerant amylases.By thermal inactivation assays, we determined that SdGA is thermolabile at temperatures above 42°C and we found that glycerol 10% (V/V), acarbose 0.1 mM and NaCl 1 M stabilized the enzyme. Furthermore, we analyze the CD of SdGA, other cold-adapted, psychrophilic and thermostable GAs and we found that SdGA has a larger CD due to various amino acid insertions and a higher content of flexible residues compared to other thermostable GAs. These characteristics of SdGA allow it to be classified as a coldadaptedenzyme but also, a salt-tolerant enzyme. We propose that this novel SdGA, might have potential applications for use in different industrial processes that require an efficient alpha glucosidase activity at low/mild temperatures, such as biofuel production.
publishDate 2021
dc.date.none.fl_str_mv 2021
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
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http://purl.org/coar/resource_type/c_5794
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status_str publishedVersion
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dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/204462
Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradans; Tercer Encuentro de Red Argentina de Tecnología Enzimática; Primer Workshop de la Red Argentina de Tecnología Enzimática; Rosario; Argentina; 2021; 61-62
CONICET Digital
CONICET
url http://hdl.handle.net/11336/204462
identifier_str_mv Characterization of SdGA, a cold-adapted and salt-tolerant glucoamylase from Saccharophagus degradans; Tercer Encuentro de Red Argentina de Tecnología Enzimática; Primer Workshop de la Red Argentina de Tecnología Enzimática; Rosario; Argentina; 2021; 61-62
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.redtez.com.ar/wp-content/uploads/WorkshopRedTEz2021_BookAbstracts.pdf
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dc.format.none.fl_str_mv application/pdf
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dc.coverage.none.fl_str_mv Internacional
dc.publisher.none.fl_str_mv Red Argentina de Tecnología Enzimática
publisher.none.fl_str_mv Red Argentina de Tecnología Enzimática
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repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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