Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coli
- Autores
- Correa Aragunde, Maria Natalia; Nejamkin, Andres; del Castello, Fiorella Paola; Foresi, Noelia Pamela; Lamattina, Lorenzo
- Año de publicación
- 2020
- Idioma
- inglés
- Tipo de recurso
- documento de conferencia
- Estado
- versión publicada
- Descripción
- Nitric oxide synthase (NOS) catalyzes the oxidation of the substrate L-Arginine (Arg) to produce citrulline and nitric oxide (NO). We have characterized NOS from two photosynthetic microorganisms: the NOS from the alga Ostreococcus tauri (OtNOS) and the cyanobacteria Synechococcus PCC 7335 (SyNOS). OtNOS and SyNOS possess distinct biochemical properties. OtNOS is a canonical NOS similar to animal NOS with an ultrafast NO producing activity. On the contrary, an extra globin domain present in SyNOS enzyme oxidizes over 70 % of the NO-produced to nitrate (NO3-). Here we describethe expression of recombinant OtNOS and SyNOS in Escherichia coli BL12 strain and analyze bacterial growth and tolerance to nitrosative stress. Results show that the E. coli cultures expressing OtNOS and SyNOS reach a higher OD at the exponential phase with respect to bacteria transformed with the empty vector (EV). This result correlates with higher NOS protein levels assayed by immunoblot, total protein and nitrate content in NOS recombinant strain cultures. Moreover, the expression of SyNOS and at less extent of OtNOS confers the ability to grow in minimal medium with Arg as a sole N source (and plenty C-source), suggesting that NOS enzymes are active in E. coli. The high NO producing activity reported in OtNOS correlates with the flavohemoglobin hmp induction in E. coli strain expressing OtNOS, suggesting that this strain senses nitrosative stress. Furthermore, nitrosative stress generated by the addition of 1 mM of the NO donor sodium nitroprusside (SNP) reduced growth rate (0.4-fold respect to no SNP addition) in bacterial culture expressing EV. However, the expression of recombinant OtNOS and at less extent SyNOS, attenuated SNP toxicity (0.8- and 0.6-fold, respectively, compared to no SNP addition). E. coli does not synthesize the major NOS cofactor tetrahydrobiopterin (BH4). Bioinformatics tools and ligand docking analysis were used to provide evidence supporting tetrahydromonapterin (MH4) as a possible pterin cofactor required for NOS catalytic activity in E. coli. These results open an exciting new window about the versatility of pterin cofactor working in the different NOSs dispersed in distant organisms along the life tree. In summary, our results show that NOS from photosynthetic microorganisms increases the growth and confers nitrosative stress tolerance in E. coli. Supported by AGENCIA, CONICET and UNMdP
Fil: Correa Aragunde, Maria Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Nejamkin, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: del Castello, Fiorella Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Foresi, Noelia Pamela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Lamattina, Lorenzo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Reunión Conjunta SAIB-SAMIGE 2020. LVI Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular. LVI Reunión Anual Sociedad Argentina de Bioquímica y Biología Molecular (SAIB). XV Reunión Anual Sociedad Argentina de Microbiología General (SAMIGE)
Buenos Aires
Argentina
Sociedad Argentina de Investigación en Bioquímica y Biología Molecular
Asociación Civil de Microbiología General - Materia
-
BACTERIA
GROWTH
NITRIC OXIDE SYNTHASE
OXIDATIVE STRESS - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/217194
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Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coliCorrea Aragunde, Maria NataliaNejamkin, Andresdel Castello, Fiorella PaolaForesi, Noelia PamelaLamattina, LorenzoBACTERIAGROWTHNITRIC OXIDE SYNTHASEOXIDATIVE STRESShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Nitric oxide synthase (NOS) catalyzes the oxidation of the substrate L-Arginine (Arg) to produce citrulline and nitric oxide (NO). We have characterized NOS from two photosynthetic microorganisms: the NOS from the alga Ostreococcus tauri (OtNOS) and the cyanobacteria Synechococcus PCC 7335 (SyNOS). OtNOS and SyNOS possess distinct biochemical properties. OtNOS is a canonical NOS similar to animal NOS with an ultrafast NO producing activity. On the contrary, an extra globin domain present in SyNOS enzyme oxidizes over 70 % of the NO-produced to nitrate (NO3-). Here we describethe expression of recombinant OtNOS and SyNOS in Escherichia coli BL12 strain and analyze bacterial growth and tolerance to nitrosative stress. Results show that the E. coli cultures expressing OtNOS and SyNOS reach a higher OD at the exponential phase with respect to bacteria transformed with the empty vector (EV). This result correlates with higher NOS protein levels assayed by immunoblot, total protein and nitrate content in NOS recombinant strain cultures. Moreover, the expression of SyNOS and at less extent of OtNOS confers the ability to grow in minimal medium with Arg as a sole N source (and plenty C-source), suggesting that NOS enzymes are active in E. coli. The high NO producing activity reported in OtNOS correlates with the flavohemoglobin hmp induction in E. coli strain expressing OtNOS, suggesting that this strain senses nitrosative stress. Furthermore, nitrosative stress generated by the addition of 1 mM of the NO donor sodium nitroprusside (SNP) reduced growth rate (0.4-fold respect to no SNP addition) in bacterial culture expressing EV. However, the expression of recombinant OtNOS and at less extent SyNOS, attenuated SNP toxicity (0.8- and 0.6-fold, respectively, compared to no SNP addition). E. coli does not synthesize the major NOS cofactor tetrahydrobiopterin (BH4). Bioinformatics tools and ligand docking analysis were used to provide evidence supporting tetrahydromonapterin (MH4) as a possible pterin cofactor required for NOS catalytic activity in E. coli. These results open an exciting new window about the versatility of pterin cofactor working in the different NOSs dispersed in distant organisms along the life tree. In summary, our results show that NOS from photosynthetic microorganisms increases the growth and confers nitrosative stress tolerance in E. coli. Supported by AGENCIA, CONICET and UNMdPFil: Correa Aragunde, Maria Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Nejamkin, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: del Castello, Fiorella Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Foresi, Noelia Pamela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Lamattina, Lorenzo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaReunión Conjunta SAIB-SAMIGE 2020. LVI Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular. LVI Reunión Anual Sociedad Argentina de Bioquímica y Biología Molecular (SAIB). XV Reunión Anual Sociedad Argentina de Microbiología General (SAMIGE)Buenos AiresArgentinaSociedad Argentina de Investigación en Bioquímica y Biología MolecularAsociación Civil de Microbiología GeneralTech Science Press2020info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectCongresoJournalhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/217194Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coli; Reunión Conjunta SAIB-SAMIGE 2020. LVI Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular. LVI Reunión Anual Sociedad Argentina de Bioquímica y Biología Molecular (SAIB). XV Reunión Anual Sociedad Argentina de Microbiología General (SAMIGE); Buenos Aires; Argentina; 2020; 108-1080327-95451667-5746CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.samige.org.ar/admin/news/files/170-TSP_BIOCELL_42376.pdfNacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:26:51Zoai:ri.conicet.gov.ar:11336/217194instacron: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 10:26:51.802CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coli |
title |
Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coli |
spellingShingle |
Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coli Correa Aragunde, Maria Natalia BACTERIA GROWTH NITRIC OXIDE SYNTHASE OXIDATIVE STRESS |
title_short |
Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coli |
title_full |
Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coli |
title_fullStr |
Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coli |
title_full_unstemmed |
Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coli |
title_sort |
Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coli |
dc.creator.none.fl_str_mv |
Correa Aragunde, Maria Natalia Nejamkin, Andres del Castello, Fiorella Paola Foresi, Noelia Pamela Lamattina, Lorenzo |
author |
Correa Aragunde, Maria Natalia |
author_facet |
Correa Aragunde, Maria Natalia Nejamkin, Andres del Castello, Fiorella Paola Foresi, Noelia Pamela Lamattina, Lorenzo |
author_role |
author |
author2 |
Nejamkin, Andres del Castello, Fiorella Paola Foresi, Noelia Pamela Lamattina, Lorenzo |
author2_role |
author author author author |
dc.subject.none.fl_str_mv |
BACTERIA GROWTH NITRIC OXIDE SYNTHASE OXIDATIVE STRESS |
topic |
BACTERIA GROWTH NITRIC OXIDE SYNTHASE OXIDATIVE STRESS |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
dc.description.none.fl_txt_mv |
Nitric oxide synthase (NOS) catalyzes the oxidation of the substrate L-Arginine (Arg) to produce citrulline and nitric oxide (NO). We have characterized NOS from two photosynthetic microorganisms: the NOS from the alga Ostreococcus tauri (OtNOS) and the cyanobacteria Synechococcus PCC 7335 (SyNOS). OtNOS and SyNOS possess distinct biochemical properties. OtNOS is a canonical NOS similar to animal NOS with an ultrafast NO producing activity. On the contrary, an extra globin domain present in SyNOS enzyme oxidizes over 70 % of the NO-produced to nitrate (NO3-). Here we describethe expression of recombinant OtNOS and SyNOS in Escherichia coli BL12 strain and analyze bacterial growth and tolerance to nitrosative stress. Results show that the E. coli cultures expressing OtNOS and SyNOS reach a higher OD at the exponential phase with respect to bacteria transformed with the empty vector (EV). This result correlates with higher NOS protein levels assayed by immunoblot, total protein and nitrate content in NOS recombinant strain cultures. Moreover, the expression of SyNOS and at less extent of OtNOS confers the ability to grow in minimal medium with Arg as a sole N source (and plenty C-source), suggesting that NOS enzymes are active in E. coli. The high NO producing activity reported in OtNOS correlates with the flavohemoglobin hmp induction in E. coli strain expressing OtNOS, suggesting that this strain senses nitrosative stress. Furthermore, nitrosative stress generated by the addition of 1 mM of the NO donor sodium nitroprusside (SNP) reduced growth rate (0.4-fold respect to no SNP addition) in bacterial culture expressing EV. However, the expression of recombinant OtNOS and at less extent SyNOS, attenuated SNP toxicity (0.8- and 0.6-fold, respectively, compared to no SNP addition). E. coli does not synthesize the major NOS cofactor tetrahydrobiopterin (BH4). Bioinformatics tools and ligand docking analysis were used to provide evidence supporting tetrahydromonapterin (MH4) as a possible pterin cofactor required for NOS catalytic activity in E. coli. These results open an exciting new window about the versatility of pterin cofactor working in the different NOSs dispersed in distant organisms along the life tree. In summary, our results show that NOS from photosynthetic microorganisms increases the growth and confers nitrosative stress tolerance in E. coli. Supported by AGENCIA, CONICET and UNMdP Fil: Correa Aragunde, Maria Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina Fil: Nejamkin, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina Fil: del Castello, Fiorella Paola. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina Fil: Foresi, Noelia Pamela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina Fil: Lamattina, Lorenzo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina Reunión Conjunta SAIB-SAMIGE 2020. LVI Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular. LVI Reunión Anual Sociedad Argentina de Bioquímica y Biología Molecular (SAIB). XV Reunión Anual Sociedad Argentina de Microbiología General (SAMIGE) Buenos Aires Argentina Sociedad Argentina de Investigación en Bioquímica y Biología Molecular Asociación Civil de Microbiología General |
description |
Nitric oxide synthase (NOS) catalyzes the oxidation of the substrate L-Arginine (Arg) to produce citrulline and nitric oxide (NO). We have characterized NOS from two photosynthetic microorganisms: the NOS from the alga Ostreococcus tauri (OtNOS) and the cyanobacteria Synechococcus PCC 7335 (SyNOS). OtNOS and SyNOS possess distinct biochemical properties. OtNOS is a canonical NOS similar to animal NOS with an ultrafast NO producing activity. On the contrary, an extra globin domain present in SyNOS enzyme oxidizes over 70 % of the NO-produced to nitrate (NO3-). Here we describethe expression of recombinant OtNOS and SyNOS in Escherichia coli BL12 strain and analyze bacterial growth and tolerance to nitrosative stress. Results show that the E. coli cultures expressing OtNOS and SyNOS reach a higher OD at the exponential phase with respect to bacteria transformed with the empty vector (EV). This result correlates with higher NOS protein levels assayed by immunoblot, total protein and nitrate content in NOS recombinant strain cultures. Moreover, the expression of SyNOS and at less extent of OtNOS confers the ability to grow in minimal medium with Arg as a sole N source (and plenty C-source), suggesting that NOS enzymes are active in E. coli. The high NO producing activity reported in OtNOS correlates with the flavohemoglobin hmp induction in E. coli strain expressing OtNOS, suggesting that this strain senses nitrosative stress. Furthermore, nitrosative stress generated by the addition of 1 mM of the NO donor sodium nitroprusside (SNP) reduced growth rate (0.4-fold respect to no SNP addition) in bacterial culture expressing EV. However, the expression of recombinant OtNOS and at less extent SyNOS, attenuated SNP toxicity (0.8- and 0.6-fold, respectively, compared to no SNP addition). E. coli does not synthesize the major NOS cofactor tetrahydrobiopterin (BH4). Bioinformatics tools and ligand docking analysis were used to provide evidence supporting tetrahydromonapterin (MH4) as a possible pterin cofactor required for NOS catalytic activity in E. coli. These results open an exciting new window about the versatility of pterin cofactor working in the different NOSs dispersed in distant organisms along the life tree. In summary, our results show that NOS from photosynthetic microorganisms increases the growth and confers nitrosative stress tolerance in E. coli. Supported by AGENCIA, CONICET and UNMdP |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020 |
dc.type.none.fl_str_mv |
info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/conferenceObject Congreso Journal http://purl.org/coar/resource_type/c_5794 info:ar-repo/semantics/documentoDeConferencia |
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publishedVersion |
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conferenceObject |
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http://hdl.handle.net/11336/217194 Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coli; Reunión Conjunta SAIB-SAMIGE 2020. LVI Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular. LVI Reunión Anual Sociedad Argentina de Bioquímica y Biología Molecular (SAIB). XV Reunión Anual Sociedad Argentina de Microbiología General (SAMIGE); Buenos Aires; Argentina; 2020; 108-108 0327-9545 1667-5746 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/217194 |
identifier_str_mv |
Expression of nitric oxide synthases from photosynthetic microorganisms improves growth and stress tolerance in E. coli; Reunión Conjunta SAIB-SAMIGE 2020. LVI Reunión Anual de la Sociedad Argentina de Investigación en Bioquímica y Biología Molecular. LVI Reunión Anual Sociedad Argentina de Bioquímica y Biología Molecular (SAIB). XV Reunión Anual Sociedad Argentina de Microbiología General (SAMIGE); Buenos Aires; Argentina; 2020; 108-108 0327-9545 1667-5746 CONICET Digital CONICET |
dc.language.none.fl_str_mv |
eng |
language |
eng |
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info:eu-repo/semantics/altIdentifier/url/https://www.samige.org.ar/admin/news/files/170-TSP_BIOCELL_42376.pdf |
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Tech Science Press |
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Tech Science Press |
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dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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