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Optimización de la expresión de la metaloproteinasa recombinante BaP1 en Pichia pastoris

Authors: Retana Lobo, Arturo José;

Optimización de la expresión de la metaloproteinasa recombinante BaP1 en Pichia pastoris

Abstract

La proteína BaP1 es una metaloproteinasa de la clase P1 del veneno de la serpiente Bothrops asper, la cual contribuye al daño tisular local al inducir inflamación, alteraciones de la matriz extracelular, mionecrosis, dermonecrosis y hemorragia local. La tecnología de ADN recombinante supone una herramienta para realizar modificaciones en su secuencia codificante de proteínas y así estudiar las relaciones entre su estructura y función. El sistema de expresión en la levadura Pichia pastoris es una alternativa rápida, poco costosa, de fácil cultivo, con disponibilidad comercial y capaz de altos niveles de proteínas expresadas con apropiadas modificaciones postraduccionales. En el presente trabajo se optimizó la cepa de Pichia pastoris, la temperatura y el pH de expresión; hallando un mayor rendimiento en la cepa KM71H que presenta un metabolismo lento del metanol (MutS) y utilizando un pH de 6. Variaciones en la temperatura permiten diferentes patrones de corte, encontrándose un predominio de la proenzima a 20°C y un predominio de la enzima madura a 30°C.

Vicerrectoría de Docencia::Salud::Facultad de Microbiología

Country
Costa Rica
Related Organizations
Keywords

modificaciones postraduccionales, pH, ADN recombinante, Bothrops asper, secuencia codificante de proteínas, X33, metaloproteniasa, SVMPs, expresión recombinante, biología molecular, KM71H, Pichia pastoris, BaP1, fenotipo Mut, metaloproteinasas de venenos de serpientes, metaloproteinasa, enzimología, tratamiento antiofídico, cepa, GS115, sistema de expresión, Metaloproteinasas de venenos de serpientes, toxinología

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Powered by OpenAIRE graph
Found an issue? Give us feedback
selected citations
These citations are derived from selected sources.
This is an alternative to the "Influence" indicator, which also reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Citations provided by BIP!
popularity
This indicator reflects the "current" impact/attention (the "hype") of an article in the research community at large, based on the underlying citation network.
BIP!Popularity provided by BIP!
influence
This indicator reflects the overall/total impact of an article in the research community at large, based on the underlying citation network (diachronically).
BIP!Influence provided by BIP!
impulse
This indicator reflects the initial momentum of an article directly after its publication, based on the underlying citation network.
BIP!Impulse provided by BIP!
0
Average
Average
Average
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