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
SEDICI (UNLP)
Institución
Universidad Nacional de La Plata
OAI Identificador
oai:sedici.unlp.edu.ar:10915/67029

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network_name_str SEDICI (UNLP)
spelling 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
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info:ar-repo/semantics/articulo
format article
status_str 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)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0)
dc.format.none.fl_str_mv application/pdf
86-96
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instname_str Universidad Nacional de La Plata
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repository.name.fl_str_mv SEDICI (UNLP) - Universidad Nacional de La Plata
repository.mail.fl_str_mv alira@sedici.unlp.edu.ar
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