Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolor

Autores
Arabolaza, Ana Lorena; Rodriguez, Eduardo Jose; Altabe, Silvia Graciela; Alvarez, Hector Manuel; Gramajo, Hugo Cesar
Año de publicación
2008
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The terminal reaction in triacylglyceride (TAG) biosynthesis is esterification of diacylglycerol (DAG) with a fatty acid molecule. To study it in Streptomyces coelicolor we analyzed three candidate genes (sco0958, sco1280, sco0123) whose products significantly resemble the recently identified wax ester synthase/acyl-CoA:diacylglycerol acyltransferase (WS/DGAT) from Acinetobacter baylyi. Deletion of either sco0123 or sco1280 resulted in no detectable decrease in TAG accumulation. In contrast, deletion of sco0958 produced a dramatic reduction in neutral lipid production, whereas over-expression of this gene yielded a significant increase in de novo TAG biosynthesis. In vitro activity assays showed that Sco0958 mediates esterification of DAG using long-chain acyl-CoAs (C14 to C18) as acyl donors. The Km and Vmax of this enzyme for myristoyl-CoA were 45 microM and 822 nmol mg(-1) min(-1), respectively. Significantly, the triple mutant strain was not completely devoid of storage lipids, indicating the existence of alternative TAG biosynthetic routes. We present strong evidence demonstrating that the residual production of TAG in this mutant strain is mediated, at least in part, by an acyl CoA-dependent pathway, since the triple mutant still exhibited DGAT activity. More importantly, there was substantial phospholipid: diacylglycerol acyltransferase activity (PDAT) in the wild type and in the triple mutant. This is the first time that a PDAT activity has been reported in bacteria, highlighting the extreme metabolic diversity of this industrially important soil microorganism.
Fil: Arabolaza, Ana Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Rodriguez, Eduardo Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Altabe, Silvia Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Alvarez, Hector Manuel. Universidad Nacional de la Patagonia "San Juan Bosco". Facultad de Ciencias Naturales - Sede Comodoro; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Gramajo, Hugo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Materia
TAG
WS/DGAT
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:11336/244790
Acceder
id CONICETDig_56fbec5c305d2066323caacfeeda8527
oai_identifier_str oai:ri.conicet.gov.ar:11336/244790
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolorArabolaza, Ana LorenaRodriguez, Eduardo JoseAltabe, Silvia GracielaAlvarez, Hector ManuelGramajo, Hugo CesarTAGWS/DGAThttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The terminal reaction in triacylglyceride (TAG) biosynthesis is esterification of diacylglycerol (DAG) with a fatty acid molecule. To study it in Streptomyces coelicolor we analyzed three candidate genes (sco0958, sco1280, sco0123) whose products significantly resemble the recently identified wax ester synthase/acyl-CoA:diacylglycerol acyltransferase (WS/DGAT) from Acinetobacter baylyi. Deletion of either sco0123 or sco1280 resulted in no detectable decrease in TAG accumulation. In contrast, deletion of sco0958 produced a dramatic reduction in neutral lipid production, whereas over-expression of this gene yielded a significant increase in de novo TAG biosynthesis. In vitro activity assays showed that Sco0958 mediates esterification of DAG using long-chain acyl-CoAs (C14 to C18) as acyl donors. The Km and Vmax of this enzyme for myristoyl-CoA were 45 microM and 822 nmol mg(-1) min(-1), respectively. Significantly, the triple mutant strain was not completely devoid of storage lipids, indicating the existence of alternative TAG biosynthetic routes. We present strong evidence demonstrating that the residual production of TAG in this mutant strain is mediated, at least in part, by an acyl CoA-dependent pathway, since the triple mutant still exhibited DGAT activity. More importantly, there was substantial phospholipid: diacylglycerol acyltransferase activity (PDAT) in the wild type and in the triple mutant. This is the first time that a PDAT activity has been reported in bacteria, highlighting the extreme metabolic diversity of this industrially important soil microorganism.Fil: Arabolaza, Ana Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Rodriguez, Eduardo Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Altabe, Silvia Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Alvarez, Hector Manuel. Universidad Nacional de la Patagonia "San Juan Bosco". Facultad de Ciencias Naturales - Sede Comodoro; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Gramajo, Hugo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaAmerican Society for Microbiology2008-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/244790Arabolaza, Ana Lorena; Rodriguez, Eduardo Jose; Altabe, Silvia Graciela; Alvarez, Hector Manuel; Gramajo, Hugo Cesar; Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolor; American Society for Microbiology; Applied and Environmental Microbiology; 74; 9; 5-2008; 2573-25820099-2240CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://journals.asm.org/doi/full/10.1128/aem.02638-07info:eu-repo/semantics/altIdentifier/doi/10.1128/AEM.02638-07info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:41:48Zoai:ri.conicet.gov.ar:11336/244790instacron: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:34982025-09-29 09:41:49.048CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolor
title Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolor
spellingShingle Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolor
Arabolaza, Ana Lorena
TAG
WS/DGAT
title_short Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolor
title_full Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolor
title_fullStr Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolor
title_full_unstemmed Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolor
title_sort Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolor
dc.creator.none.fl_str_mv Arabolaza, Ana Lorena
Rodriguez, Eduardo Jose
Altabe, Silvia Graciela
Alvarez, Hector Manuel
Gramajo, Hugo Cesar
author Arabolaza, Ana Lorena
author_facet Arabolaza, Ana Lorena
Rodriguez, Eduardo Jose
Altabe, Silvia Graciela
Alvarez, Hector Manuel
Gramajo, Hugo Cesar
author_role author
author2 Rodriguez, Eduardo Jose
Altabe, Silvia Graciela
Alvarez, Hector Manuel
Gramajo, Hugo Cesar
author2_role author
author
author
author
dc.subject.none.fl_str_mv TAG
WS/DGAT
topic TAG
WS/DGAT
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 terminal reaction in triacylglyceride (TAG) biosynthesis is esterification of diacylglycerol (DAG) with a fatty acid molecule. To study it in Streptomyces coelicolor we analyzed three candidate genes (sco0958, sco1280, sco0123) whose products significantly resemble the recently identified wax ester synthase/acyl-CoA:diacylglycerol acyltransferase (WS/DGAT) from Acinetobacter baylyi. Deletion of either sco0123 or sco1280 resulted in no detectable decrease in TAG accumulation. In contrast, deletion of sco0958 produced a dramatic reduction in neutral lipid production, whereas over-expression of this gene yielded a significant increase in de novo TAG biosynthesis. In vitro activity assays showed that Sco0958 mediates esterification of DAG using long-chain acyl-CoAs (C14 to C18) as acyl donors. The Km and Vmax of this enzyme for myristoyl-CoA were 45 microM and 822 nmol mg(-1) min(-1), respectively. Significantly, the triple mutant strain was not completely devoid of storage lipids, indicating the existence of alternative TAG biosynthetic routes. We present strong evidence demonstrating that the residual production of TAG in this mutant strain is mediated, at least in part, by an acyl CoA-dependent pathway, since the triple mutant still exhibited DGAT activity. More importantly, there was substantial phospholipid: diacylglycerol acyltransferase activity (PDAT) in the wild type and in the triple mutant. This is the first time that a PDAT activity has been reported in bacteria, highlighting the extreme metabolic diversity of this industrially important soil microorganism.
Fil: Arabolaza, Ana Lorena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Rodriguez, Eduardo Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Altabe, Silvia Graciela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
Fil: Alvarez, Hector Manuel. Universidad Nacional de la Patagonia "San Juan Bosco". Facultad de Ciencias Naturales - Sede Comodoro; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Gramajo, Hugo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; Argentina
description The terminal reaction in triacylglyceride (TAG) biosynthesis is esterification of diacylglycerol (DAG) with a fatty acid molecule. To study it in Streptomyces coelicolor we analyzed three candidate genes (sco0958, sco1280, sco0123) whose products significantly resemble the recently identified wax ester synthase/acyl-CoA:diacylglycerol acyltransferase (WS/DGAT) from Acinetobacter baylyi. Deletion of either sco0123 or sco1280 resulted in no detectable decrease in TAG accumulation. In contrast, deletion of sco0958 produced a dramatic reduction in neutral lipid production, whereas over-expression of this gene yielded a significant increase in de novo TAG biosynthesis. In vitro activity assays showed that Sco0958 mediates esterification of DAG using long-chain acyl-CoAs (C14 to C18) as acyl donors. The Km and Vmax of this enzyme for myristoyl-CoA were 45 microM and 822 nmol mg(-1) min(-1), respectively. Significantly, the triple mutant strain was not completely devoid of storage lipids, indicating the existence of alternative TAG biosynthetic routes. We present strong evidence demonstrating that the residual production of TAG in this mutant strain is mediated, at least in part, by an acyl CoA-dependent pathway, since the triple mutant still exhibited DGAT activity. More importantly, there was substantial phospholipid: diacylglycerol acyltransferase activity (PDAT) in the wild type and in the triple mutant. This is the first time that a PDAT activity has been reported in bacteria, highlighting the extreme metabolic diversity of this industrially important soil microorganism.
publishDate 2008
dc.date.none.fl_str_mv 2008-05
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/11336/244790
Arabolaza, Ana Lorena; Rodriguez, Eduardo Jose; Altabe, Silvia Graciela; Alvarez, Hector Manuel; Gramajo, Hugo Cesar; Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolor; American Society for Microbiology; Applied and Environmental Microbiology; 74; 9; 5-2008; 2573-2582
0099-2240
CONICET Digital
CONICET
url http://hdl.handle.net/11336/244790
identifier_str_mv Arabolaza, Ana Lorena; Rodriguez, Eduardo Jose; Altabe, Silvia Graciela; Alvarez, Hector Manuel; Gramajo, Hugo Cesar; Multiple Pathways for Triacylglycerol Biosynthesis in Streptomyces coelicolor; American Society for Microbiology; Applied and Environmental Microbiology; 74; 9; 5-2008; 2573-2582
0099-2240
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://journals.asm.org/doi/full/10.1128/aem.02638-07
info:eu-repo/semantics/altIdentifier/doi/10.1128/AEM.02638-07
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
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
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 13.070432

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