Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in <italic toggle="yes">Streptomyces coelicolor</italic> M145

ABSTRACT Streptomyces coelicolor is a Gram-positive soil bacterium with a high metabolic and adaptive potential that is able to utilize a variety of nitrogen sources. However, little is known about the utilization of the alternative nitrogen source ethanolamine. Our study revealed that S. coelicolor...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Sergii Krysenko, Arne Matthews, Nicole Okoniewski, Andreas Kulik, Melis G. Girbas, Olga Tsypik, Christian Stephan Meyners, Felix Hausch, Wolfgang Wohlleben, Agnieszka Bera
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Streptomyces ethanolamine utilization gamma-glutamylation nitrogen metabolism Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/01dd08b98bc744a2b1d7d75139ece7d2
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:01dd08b98bc744a2b1d7d75139ece7d2
record_format	dspace
spelling	oai:doaj.org-article:01dd08b98bc744a2b1d7d75139ece7d22021-11-15T15:55:24ZInitial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in <italic toggle="yes">Streptomyces coelicolor</italic> M14510.1128/mBio.00326-192150-7511https://doaj.org/article/01dd08b98bc744a2b1d7d75139ece7d22019-06-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00326-19https://doaj.org/toc/2150-7511ABSTRACT Streptomyces coelicolor is a Gram-positive soil bacterium with a high metabolic and adaptive potential that is able to utilize a variety of nitrogen sources. However, little is known about the utilization of the alternative nitrogen source ethanolamine. Our study revealed that S. coelicolor can utilize ethanolamine as a sole nitrogen or carbon (N/C) source, although it grows poorly on this nitrogen source due to the absence of a specific ethanolamine permease. Heterologous expression of a putative ethanolamine permease (SPRI_5940) from Streptomycespristinaespiralis positively influenced the biomass accumulation of the overexpression strain grown in defined medium with ethanolamine. In this study, we demonstrated that a glutamine synthetase-like protein, GlnA4 (SCO1613), is involved in the initial metabolic step of a novel ethanolamine utilization pathway in S. coelicolor M145. GlnA4 acts as a gamma-glutamylethanolamide synthetase. Transcriptional analysis revealed that expression of glnA4 was induced by ethanolamine and repressed in the presence of ammonium. Regulation of glnA4 is governed by the transcriptional repressor EpuRI (SCO1614). The ΔglnA4 mutant strain was unable to grow on defined liquid Evans medium supplemented with ethanolamine. High-performance liquid chromatography (HPLC) analysis demonstrated that strain ΔglnA4 is unable to utilize ethanolamine. GlnA4-catalyzed glutamylation of ethanolamine was confirmed in an enzymatic in vitro assay, and the GlnA4 reaction product, gamma-glutamylethanolamide, was detected by HPLC/electrospray ionization-mass spectrometry (HPLC/ESI-MS). In this work, the first step of ethanolamine utilization in S. coelicolor M145 was elucidated, and a putative ethanolamine utilization pathway was deduced based on the sequence similarity and genomic localization of homologous genes. IMPORTANCE Until now, knowledge of the utilization of ethanolamine in Streptomyces was limited. Our work represents the first attempt to reveal a novel ethanolamine utilization pathway in the actinobacterial model organism S. coelicolor through the characterization of the key enzyme gamma-glutamylethanolamide synthetase GlnA4, which is absolutely required for growth in the presence of ethanolamine. The novel ethanolamine utilization pathway is dissimilar to the currently known ethanolamine utilization pathway, which occurs in metabolome. The novel ethanolamine utilization pathway does not result in the production of toxic by-products (such as acetaldehyde); thus, it is not encapsulated. We believe that this contribution is a milestone in understanding the ecology of Streptomyces and the utilization of alternative nitrogen sources. Our report provides new insight into bacterial primary metabolism, which remains complex and partially unexplored.Sergii KrysenkoArne MatthewsNicole OkoniewskiAndreas KulikMelis G. GirbasOlga TsypikChristian Stephan MeynersFelix HauschWolfgang WohllebenAgnieszka BeraAmerican Society for MicrobiologyarticleStreptomycesethanolamine utilizationgamma-glutamylationnitrogen metabolismMicrobiologyQR1-502ENmBio, Vol 10, Iss 3 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Streptomyces ethanolamine utilization gamma-glutamylation nitrogen metabolism Microbiology QR1-502
spellingShingle	Streptomyces ethanolamine utilization gamma-glutamylation nitrogen metabolism Microbiology QR1-502 Sergii Krysenko Arne Matthews Nicole Okoniewski Andreas Kulik Melis G. Girbas Olga Tsypik Christian Stephan Meyners Felix Hausch Wolfgang Wohlleben Agnieszka Bera Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in <italic toggle="yes">Streptomyces coelicolor</italic> M145
description	ABSTRACT Streptomyces coelicolor is a Gram-positive soil bacterium with a high metabolic and adaptive potential that is able to utilize a variety of nitrogen sources. However, little is known about the utilization of the alternative nitrogen source ethanolamine. Our study revealed that S. coelicolor can utilize ethanolamine as a sole nitrogen or carbon (N/C) source, although it grows poorly on this nitrogen source due to the absence of a specific ethanolamine permease. Heterologous expression of a putative ethanolamine permease (SPRI_5940) from Streptomycespristinaespiralis positively influenced the biomass accumulation of the overexpression strain grown in defined medium with ethanolamine. In this study, we demonstrated that a glutamine synthetase-like protein, GlnA4 (SCO1613), is involved in the initial metabolic step of a novel ethanolamine utilization pathway in S. coelicolor M145. GlnA4 acts as a gamma-glutamylethanolamide synthetase. Transcriptional analysis revealed that expression of glnA4 was induced by ethanolamine and repressed in the presence of ammonium. Regulation of glnA4 is governed by the transcriptional repressor EpuRI (SCO1614). The ΔglnA4 mutant strain was unable to grow on defined liquid Evans medium supplemented with ethanolamine. High-performance liquid chromatography (HPLC) analysis demonstrated that strain ΔglnA4 is unable to utilize ethanolamine. GlnA4-catalyzed glutamylation of ethanolamine was confirmed in an enzymatic in vitro assay, and the GlnA4 reaction product, gamma-glutamylethanolamide, was detected by HPLC/electrospray ionization-mass spectrometry (HPLC/ESI-MS). In this work, the first step of ethanolamine utilization in S. coelicolor M145 was elucidated, and a putative ethanolamine utilization pathway was deduced based on the sequence similarity and genomic localization of homologous genes. IMPORTANCE Until now, knowledge of the utilization of ethanolamine in Streptomyces was limited. Our work represents the first attempt to reveal a novel ethanolamine utilization pathway in the actinobacterial model organism S. coelicolor through the characterization of the key enzyme gamma-glutamylethanolamide synthetase GlnA4, which is absolutely required for growth in the presence of ethanolamine. The novel ethanolamine utilization pathway is dissimilar to the currently known ethanolamine utilization pathway, which occurs in metabolome. The novel ethanolamine utilization pathway does not result in the production of toxic by-products (such as acetaldehyde); thus, it is not encapsulated. We believe that this contribution is a milestone in understanding the ecology of Streptomyces and the utilization of alternative nitrogen sources. Our report provides new insight into bacterial primary metabolism, which remains complex and partially unexplored.
format	article
author	Sergii Krysenko Arne Matthews Nicole Okoniewski Andreas Kulik Melis G. Girbas Olga Tsypik Christian Stephan Meyners Felix Hausch Wolfgang Wohlleben Agnieszka Bera
author_facet	Sergii Krysenko Arne Matthews Nicole Okoniewski Andreas Kulik Melis G. Girbas Olga Tsypik Christian Stephan Meyners Felix Hausch Wolfgang Wohlleben Agnieszka Bera
author_sort	Sergii Krysenko
title	Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in <italic toggle="yes">Streptomyces coelicolor</italic> M145
title_short	Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in <italic toggle="yes">Streptomyces coelicolor</italic> M145
title_full	Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in <italic toggle="yes">Streptomyces coelicolor</italic> M145
title_fullStr	Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in <italic toggle="yes">Streptomyces coelicolor</italic> M145
title_full_unstemmed	Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in <italic toggle="yes">Streptomyces coelicolor</italic> M145
title_sort	initial metabolic step of a novel ethanolamine utilization pathway and its regulation in <italic toggle="yes">streptomyces coelicolor</italic> m145
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/01dd08b98bc744a2b1d7d75139ece7d2
work_keys_str_mv	AT sergiikrysenko initialmetabolicstepofanovelethanolamineutilizationpathwayanditsregulationinitalictoggleyesstreptomycescoelicoloritalicm145 AT arnematthews initialmetabolicstepofanovelethanolamineutilizationpathwayanditsregulationinitalictoggleyesstreptomycescoelicoloritalicm145 AT nicoleokoniewski initialmetabolicstepofanovelethanolamineutilizationpathwayanditsregulationinitalictoggleyesstreptomycescoelicoloritalicm145 AT andreaskulik initialmetabolicstepofanovelethanolamineutilizationpathwayanditsregulationinitalictoggleyesstreptomycescoelicoloritalicm145 AT melisggirbas initialmetabolicstepofanovelethanolamineutilizationpathwayanditsregulationinitalictoggleyesstreptomycescoelicoloritalicm145 AT olgatsypik initialmetabolicstepofanovelethanolamineutilizationpathwayanditsregulationinitalictoggleyesstreptomycescoelicoloritalicm145 AT christianstephanmeyners initialmetabolicstepofanovelethanolamineutilizationpathwayanditsregulationinitalictoggleyesstreptomycescoelicoloritalicm145 AT felixhausch initialmetabolicstepofanovelethanolamineutilizationpathwayanditsregulationinitalictoggleyesstreptomycescoelicoloritalicm145 AT wolfgangwohlleben initialmetabolicstepofanovelethanolamineutilizationpathwayanditsregulationinitalictoggleyesstreptomycescoelicoloritalicm145 AT agnieszkabera initialmetabolicstepofanovelethanolamineutilizationpathwayanditsregulationinitalictoggleyesstreptomycescoelicoloritalicm145
_version_	1718427193683476480

Initial Metabolic Step of a Novel Ethanolamine Utilization Pathway and Its Regulation in <italic toggle="yes">Streptomyces coelicolor</italic> M145

Ejemplares similares