Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1

Autores
Cereijo, Antonela Estefanía; Ferretti, María Victoria; Iglesias, Alberto Alvaro; Alvarez, Hector Manuel; Asención Diez, Matías Damián
Año de publicación
2023
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The bacterial genus Rhodococcus comprises organisms that perform an oleaginous behavior under certain growth conditions and the ratio of carbon and nitrogen availability. Thus, Rhodococcus spp have outstanding biotechnological features as microbial producers of biofuel precursors, which would be used instead of lipids from crops. It was postulated that lipid and glycogen metabolism in Rhodococci are closely related. Thus, a better understanding of rhodococcal carbon partitioning requires identifying the catalytic steps redirecting sugar moieties to temporal storage molecules, such as glycogen and trehalose. In this work, we analyzed two glycosyl-transferases GT4 from R. jostii, RjoGlgAb and RjoGlgAc, which were annotated as α-glucan-α-1,4-glucosyl transferases, putatively involved in glycogen synthesis. Both enzymes were recombinantly produced in E. coli BL21 (DE3) cells, purified to near homogeneity, and kinetically characterized. RjoGlgAb and RjoGlgAc presented the “canonical” glycogen synthase (EC 2.4.1.21) activity. Besides, both enzymes were actives as maltose-1P synthases (GlgM, EC 2.4.1.342), although to a different extent. In this scenario, RjoGlgAc is a homologous enzyme to the mycobacterial GlgM, with similar behavior regarding kinetic parameters and glucosyl-donor (ADP-glucose) preference. RjoGlgAc was two orders of magnitude more efficient to glucosylate glucose-1P than glycogen. Also, this rhodococcal enzyme used glucosamine-1P as a catalytically efficient aglycon. On the other hand, both activities exhibited by RjoGlgAb depicted similar kinetic efficiency and a preference for short-branched α-1,4-glucans. Curiously, RjoGlgAb presented a super-oligomeric conformation (higher than 15 subunits), representing a novel enzyme with a unique structure to function relationships. Results presented herein constitute a milestone regarding polysaccharide biosynthesis in Actinobacteria, leading to (re)discovery of methyl-glucose lipo-polysaccharide metabolism in Rhodococci.
Fil: Cereijo, Antonela Estefanía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Ferretti, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Alvarez, Hector Manuel. Universidad Nacional de la Patagonia "San Juan Bosco". Instituto de Biociencias de la Patagonia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biociencias de la Patagonia; Argentina
Fil: Asención Diez, Matías Damián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Materia
METHYL-GLUCOSE LIPOPOLYSACCHARIDE
GLUCOSAMINE-1P
GLYCOGEN
LIPIDS
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:11336/226735
Acceder
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network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1Cereijo, Antonela EstefaníaFerretti, María VictoriaIglesias, Alberto AlvaroAlvarez, Hector ManuelAsención Diez, Matías DamiánMETHYL-GLUCOSE LIPOPOLYSACCHARIDEGLUCOSAMINE-1PGLYCOGENLIPIDShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The bacterial genus Rhodococcus comprises organisms that perform an oleaginous behavior under certain growth conditions and the ratio of carbon and nitrogen availability. Thus, Rhodococcus spp have outstanding biotechnological features as microbial producers of biofuel precursors, which would be used instead of lipids from crops. It was postulated that lipid and glycogen metabolism in Rhodococci are closely related. Thus, a better understanding of rhodococcal carbon partitioning requires identifying the catalytic steps redirecting sugar moieties to temporal storage molecules, such as glycogen and trehalose. In this work, we analyzed two glycosyl-transferases GT4 from R. jostii, RjoGlgAb and RjoGlgAc, which were annotated as α-glucan-α-1,4-glucosyl transferases, putatively involved in glycogen synthesis. Both enzymes were recombinantly produced in E. coli BL21 (DE3) cells, purified to near homogeneity, and kinetically characterized. RjoGlgAb and RjoGlgAc presented the “canonical” glycogen synthase (EC 2.4.1.21) activity. Besides, both enzymes were actives as maltose-1P synthases (GlgM, EC 2.4.1.342), although to a different extent. In this scenario, RjoGlgAc is a homologous enzyme to the mycobacterial GlgM, with similar behavior regarding kinetic parameters and glucosyl-donor (ADP-glucose) preference. RjoGlgAc was two orders of magnitude more efficient to glucosylate glucose-1P than glycogen. Also, this rhodococcal enzyme used glucosamine-1P as a catalytically efficient aglycon. On the other hand, both activities exhibited by RjoGlgAb depicted similar kinetic efficiency and a preference for short-branched α-1,4-glucans. Curiously, RjoGlgAb presented a super-oligomeric conformation (higher than 15 subunits), representing a novel enzyme with a unique structure to function relationships. Results presented herein constitute a milestone regarding polysaccharide biosynthesis in Actinobacteria, leading to (re)discovery of methyl-glucose lipo-polysaccharide metabolism in Rhodococci.Fil: Cereijo, Antonela Estefanía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; ArgentinaFil: Ferretti, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; ArgentinaFil: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; ArgentinaFil: Alvarez, Hector Manuel. Universidad Nacional de la Patagonia "San Juan Bosco". Instituto de Biociencias de la Patagonia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biociencias de la Patagonia; ArgentinaFil: Asención Diez, Matías Damián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; ArgentinaCold Spring Harbor Laboratory Press2023-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/226735Cereijo, Antonela Estefanía; Ferretti, María Victoria; Iglesias, Alberto Alvaro; Alvarez, Hector Manuel; Asención Diez, Matías Damián; Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1; Cold Spring Harbor Laboratory Press; Bioxriv; 2023; 2-2023; 1-492692-8205CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.1101/2023.02.13.525685v1info:eu-repo/semantics/altIdentifier/doi/10.1101/2023.02.13.525685info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-10T13:00:57Zoai:ri.conicet.gov.ar:11336/226735instacron: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 13:00:57.608CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1
title Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1
spellingShingle Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1
Cereijo, Antonela Estefanía
METHYL-GLUCOSE LIPOPOLYSACCHARIDE
GLUCOSAMINE-1P
GLYCOGEN
LIPIDS
title_short Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1
title_full Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1
title_fullStr Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1
title_full_unstemmed Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1
title_sort Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1
dc.creator.none.fl_str_mv Cereijo, Antonela Estefanía
Ferretti, María Victoria
Iglesias, Alberto Alvaro
Alvarez, Hector Manuel
Asención Diez, Matías Damián
author Cereijo, Antonela Estefanía
author_facet Cereijo, Antonela Estefanía
Ferretti, María Victoria
Iglesias, Alberto Alvaro
Alvarez, Hector Manuel
Asención Diez, Matías Damián
author_role author
author2 Ferretti, María Victoria
Iglesias, Alberto Alvaro
Alvarez, Hector Manuel
Asención Diez, Matías Damián
author2_role author
author
author
author
dc.subject.none.fl_str_mv METHYL-GLUCOSE LIPOPOLYSACCHARIDE
GLUCOSAMINE-1P
GLYCOGEN
LIPIDS
topic METHYL-GLUCOSE LIPOPOLYSACCHARIDE
GLUCOSAMINE-1P
GLYCOGEN
LIPIDS
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 bacterial genus Rhodococcus comprises organisms that perform an oleaginous behavior under certain growth conditions and the ratio of carbon and nitrogen availability. Thus, Rhodococcus spp have outstanding biotechnological features as microbial producers of biofuel precursors, which would be used instead of lipids from crops. It was postulated that lipid and glycogen metabolism in Rhodococci are closely related. Thus, a better understanding of rhodococcal carbon partitioning requires identifying the catalytic steps redirecting sugar moieties to temporal storage molecules, such as glycogen and trehalose. In this work, we analyzed two glycosyl-transferases GT4 from R. jostii, RjoGlgAb and RjoGlgAc, which were annotated as α-glucan-α-1,4-glucosyl transferases, putatively involved in glycogen synthesis. Both enzymes were recombinantly produced in E. coli BL21 (DE3) cells, purified to near homogeneity, and kinetically characterized. RjoGlgAb and RjoGlgAc presented the “canonical” glycogen synthase (EC 2.4.1.21) activity. Besides, both enzymes were actives as maltose-1P synthases (GlgM, EC 2.4.1.342), although to a different extent. In this scenario, RjoGlgAc is a homologous enzyme to the mycobacterial GlgM, with similar behavior regarding kinetic parameters and glucosyl-donor (ADP-glucose) preference. RjoGlgAc was two orders of magnitude more efficient to glucosylate glucose-1P than glycogen. Also, this rhodococcal enzyme used glucosamine-1P as a catalytically efficient aglycon. On the other hand, both activities exhibited by RjoGlgAb depicted similar kinetic efficiency and a preference for short-branched α-1,4-glucans. Curiously, RjoGlgAb presented a super-oligomeric conformation (higher than 15 subunits), representing a novel enzyme with a unique structure to function relationships. Results presented herein constitute a milestone regarding polysaccharide biosynthesis in Actinobacteria, leading to (re)discovery of methyl-glucose lipo-polysaccharide metabolism in Rhodococci.
Fil: Cereijo, Antonela Estefanía. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Ferretti, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Iglesias, Alberto Alvaro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
Fil: Alvarez, Hector Manuel. Universidad Nacional de la Patagonia "San Juan Bosco". Instituto de Biociencias de la Patagonia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Centro Nacional Patagónico. Instituto de Biociencias de la Patagonia; Argentina
Fil: Asención Diez, Matías Damián. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Agrobiotecnología del Litoral. Universidad Nacional del Litoral. Instituto de Agrobiotecnología del Litoral; Argentina
description The bacterial genus Rhodococcus comprises organisms that perform an oleaginous behavior under certain growth conditions and the ratio of carbon and nitrogen availability. Thus, Rhodococcus spp have outstanding biotechnological features as microbial producers of biofuel precursors, which would be used instead of lipids from crops. It was postulated that lipid and glycogen metabolism in Rhodococci are closely related. Thus, a better understanding of rhodococcal carbon partitioning requires identifying the catalytic steps redirecting sugar moieties to temporal storage molecules, such as glycogen and trehalose. In this work, we analyzed two glycosyl-transferases GT4 from R. jostii, RjoGlgAb and RjoGlgAc, which were annotated as α-glucan-α-1,4-glucosyl transferases, putatively involved in glycogen synthesis. Both enzymes were recombinantly produced in E. coli BL21 (DE3) cells, purified to near homogeneity, and kinetically characterized. RjoGlgAb and RjoGlgAc presented the “canonical” glycogen synthase (EC 2.4.1.21) activity. Besides, both enzymes were actives as maltose-1P synthases (GlgM, EC 2.4.1.342), although to a different extent. In this scenario, RjoGlgAc is a homologous enzyme to the mycobacterial GlgM, with similar behavior regarding kinetic parameters and glucosyl-donor (ADP-glucose) preference. RjoGlgAc was two orders of magnitude more efficient to glucosylate glucose-1P than glycogen. Also, this rhodococcal enzyme used glucosamine-1P as a catalytically efficient aglycon. On the other hand, both activities exhibited by RjoGlgAb depicted similar kinetic efficiency and a preference for short-branched α-1,4-glucans. Curiously, RjoGlgAb presented a super-oligomeric conformation (higher than 15 subunits), representing a novel enzyme with a unique structure to function relationships. Results presented herein constitute a milestone regarding polysaccharide biosynthesis in Actinobacteria, leading to (re)discovery of methyl-glucose lipo-polysaccharide metabolism in Rhodococci.
publishDate 2023
dc.date.none.fl_str_mv 2023-02
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/11336/226735
Cereijo, Antonela Estefanía; Ferretti, María Victoria; Iglesias, Alberto Alvaro; Alvarez, Hector Manuel; Asención Diez, Matías Damián; Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1; Cold Spring Harbor Laboratory Press; Bioxriv; 2023; 2-2023; 1-49
2692-8205
CONICET Digital
CONICET
url http://hdl.handle.net/11336/226735
identifier_str_mv Cereijo, Antonela Estefanía; Ferretti, María Victoria; Iglesias, Alberto Alvaro; Alvarez, Hector Manuel; Asención Diez, Matías Damián; Comparative analysis between two GT4 glycosyltransferases related to polysaccharide biosynthesis in Rhodococcus jostii RHA1; Cold Spring Harbor Laboratory Press; Bioxriv; 2023; 2-2023; 1-49
2692-8205
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.biorxiv.org/content/10.1101/2023.02.13.525685v1
info:eu-repo/semantics/altIdentifier/doi/10.1101/2023.02.13.525685
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
dc.publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
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
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score 12.48226

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