Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin

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 an...

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Autores principales: 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
Formato: Articulo
Lenguaje:Inglés
Publicado: 2014
Materias:
Ciencias Exactas
Levaduras
Proteasas de Ácido Aspártico
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/67029
http://aem.asm.org/content/80/1/86.full
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-67029
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Exactas
Levaduras
Proteasas de Ácido Aspártico
spellingShingle Ciencias Exactas
Levaduras
Proteasas de Ácido Aspártico
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
Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin
topic_facet Ciencias Exactas
Levaduras
Proteasas de Ácido Aspártico
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.
format Articulo
Articulo
author 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_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_sort Curto, Pedro
title Establishing the Yeast <i>Kluyveromyces lactis</i> as an Expression Host for Production of the Saposin-Like Domain of the Aspartic Protease Cirsin
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
publishDate 2014
url http://sedici.unlp.edu.ar/handle/10915/67029
http://aem.asm.org/content/80/1/86.full
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