Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin
- Autores
- Curto, Pedro; Lufrano, Daniela; Pinto, Cátia; Custódio, Valeria; Gomes, Ana Catarina; Trejo, Sebastián Alejandro; Bakás, Laura Susana; Vairo Cavalli, Sandra Elizabeth; Faro, Carlos; Simões, Isaura
- Año de publicación
- 2014
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- Typical plant aspartic protease zymogens comprise a characteristic and plant-specific insert (PSI). PSI domains can interact with membranes, and a role as a defensive weapon against pathogens has been proposed. However, the potential of PSIs as antimicrobial agents has not been fully investigated and explored yet due to problems in producing sufficient amounts of these domains in bacteria. Here, we report the development of an expression platform for the production of the PSI domain of cirsin in the generally regarded as safe (GRAS) yeast Kluyveromyces lactis. We successfully generated K. lactis transformants expressing and secreting significant amounts of correctly processed and glycosylated PSI, as well as its nonglycosylated mutant. A purification protocol with protein yields of;4.0 mg/liter was established for both wild-type and nonglycosylated PSIs, which represents the highest reported yield for a nontagged PSI domain. Subsequent bioactivity assays targeting phytopathogenic fungi indicated that the PSI of cirsin is produced in a biologically active form in K. lactis and provided clear evidence for its antifungal activity. This yeast expression system thereby emerges as a promising production platform for further exploring the biotechnological potential of these plant saposin-like proteins.
Facultad de Ciencias Exactas - Materia
-
Ciencias Exactas
Levaduras
Proteasas de Ácido Aspártico - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- http://creativecommons.org/licenses/by-nc-nd/4.0/
- Repositorio
- Institución
- Universidad Nacional de La Plata
- OAI Identificador
- oai:sedici.unlp.edu.ar:10915/67029
Ver los metadatos del registro completo
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Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease CirsinCurto, PedroLufrano, DanielaPinto, CátiaCustódio, ValeriaGomes, Ana CatarinaTrejo, Sebastián AlejandroBakás, Laura SusanaVairo Cavalli, Sandra ElizabethFaro, CarlosSimões, IsauraCiencias ExactasLevadurasProteasas de Ácido AspárticoTypical plant aspartic protease zymogens comprise a characteristic and plant-specific insert (PSI). PSI domains can interact with membranes, and a role as a defensive weapon against pathogens has been proposed. However, the potential of PSIs as antimicrobial agents has not been fully investigated and explored yet due to problems in producing sufficient amounts of these domains in bacteria. Here, we report the development of an expression platform for the production of the PSI domain of cirsin in the generally regarded as safe (GRAS) yeast Kluyveromyces lactis. We successfully generated K. lactis transformants expressing and secreting significant amounts of correctly processed and glycosylated PSI, as well as its nonglycosylated mutant. A purification protocol with protein yields of;4.0 mg/liter was established for both wild-type and nonglycosylated PSIs, which represents the highest reported yield for a nontagged PSI domain. Subsequent bioactivity assays targeting phytopathogenic fungi indicated that the PSI of cirsin is produced in a biologically active form in K. lactis and provided clear evidence for its antifungal activity. This yeast expression system thereby emerges as a promising production platform for further exploring the biotechnological potential of these plant saposin-like proteins.Facultad de Ciencias Exactas2014info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdf86-96http://sedici.unlp.edu.ar/handle/10915/67029enginfo:eu-repo/semantics/altIdentifier/url/http://aem.asm.org/content/80/1/86.fullinfo:eu-repo/semantics/altIdentifier/issn/1098-5336info:eu-repo/semantics/altIdentifier/doi/10.1128/AEM.03151-13info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-nd/4.0/Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2025-09-29T11:10:10Zoai:sedici.unlp.edu.ar:10915/67029Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-29 11:10:10.495SEDICI (UNLP) - Universidad Nacional de La Platafalse |
dc.title.none.fl_str_mv |
Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin |
title |
Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin |
spellingShingle |
Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin Curto, Pedro Ciencias Exactas Levaduras Proteasas de Ácido Aspártico |
title_short |
Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin |
title_full |
Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin |
title_fullStr |
Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin |
title_full_unstemmed |
Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin |
title_sort |
Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin |
dc.creator.none.fl_str_mv |
Curto, Pedro Lufrano, Daniela Pinto, Cátia Custódio, Valeria Gomes, Ana Catarina Trejo, Sebastián Alejandro Bakás, Laura Susana Vairo Cavalli, Sandra Elizabeth Faro, Carlos Simões, Isaura |
author |
Curto, Pedro |
author_facet |
Curto, Pedro Lufrano, Daniela Pinto, Cátia Custódio, Valeria Gomes, Ana Catarina Trejo, Sebastián Alejandro Bakás, Laura Susana Vairo Cavalli, Sandra Elizabeth Faro, Carlos Simões, Isaura |
author_role |
author |
author2 |
Lufrano, Daniela Pinto, Cátia Custódio, Valeria Gomes, Ana Catarina Trejo, Sebastián Alejandro Bakás, Laura Susana Vairo Cavalli, Sandra Elizabeth Faro, Carlos Simões, Isaura |
author2_role |
author author author author author author author author author |
dc.subject.none.fl_str_mv |
Ciencias Exactas Levaduras Proteasas de Ácido Aspártico |
topic |
Ciencias Exactas Levaduras Proteasas de Ácido Aspártico |
dc.description.none.fl_txt_mv |
Typical plant aspartic protease zymogens comprise a characteristic and plant-specific insert (PSI). PSI domains can interact with membranes, and a role as a defensive weapon against pathogens has been proposed. However, the potential of PSIs as antimicrobial agents has not been fully investigated and explored yet due to problems in producing sufficient amounts of these domains in bacteria. Here, we report the development of an expression platform for the production of the PSI domain of cirsin in the generally regarded as safe (GRAS) yeast Kluyveromyces lactis. We successfully generated K. lactis transformants expressing and secreting significant amounts of correctly processed and glycosylated PSI, as well as its nonglycosylated mutant. A purification protocol with protein yields of;4.0 mg/liter was established for both wild-type and nonglycosylated PSIs, which represents the highest reported yield for a nontagged PSI domain. Subsequent bioactivity assays targeting phytopathogenic fungi indicated that the PSI of cirsin is produced in a biologically active form in K. lactis and provided clear evidence for its antifungal activity. This yeast expression system thereby emerges as a promising production platform for further exploring the biotechnological potential of these plant saposin-like proteins. Facultad de Ciencias Exactas |
description |
Typical plant aspartic protease zymogens comprise a characteristic and plant-specific insert (PSI). PSI domains can interact with membranes, and a role as a defensive weapon against pathogens has been proposed. However, the potential of PSIs as antimicrobial agents has not been fully investigated and explored yet due to problems in producing sufficient amounts of these domains in bacteria. Here, we report the development of an expression platform for the production of the PSI domain of cirsin in the generally regarded as safe (GRAS) yeast Kluyveromyces lactis. We successfully generated K. lactis transformants expressing and secreting significant amounts of correctly processed and glycosylated PSI, as well as its nonglycosylated mutant. A purification protocol with protein yields of;4.0 mg/liter was established for both wild-type and nonglycosylated PSIs, which represents the highest reported yield for a nontagged PSI domain. Subsequent bioactivity assays targeting phytopathogenic fungi indicated that the PSI of cirsin is produced in a biologically active form in K. lactis and provided clear evidence for its antifungal activity. This yeast expression system thereby emerges as a promising production platform for further exploring the biotechnological potential of these plant saposin-like proteins. |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014 |
dc.type.none.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Articulo http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
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article |
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publishedVersion |
dc.identifier.none.fl_str_mv |
http://sedici.unlp.edu.ar/handle/10915/67029 |
url |
http://sedici.unlp.edu.ar/handle/10915/67029 |
dc.language.none.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
info:eu-repo/semantics/altIdentifier/url/http://aem.asm.org/content/80/1/86.full info:eu-repo/semantics/altIdentifier/issn/1098-5336 info:eu-repo/semantics/altIdentifier/doi/10.1128/AEM.03151-13 |
dc.rights.none.fl_str_mv |
info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/4.0/ Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) |
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openAccess |
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http://creativecommons.org/licenses/by-nc-nd/4.0/ Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) |
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