Your browser doesn't support javascript.
loading
Thermal Stabilization of a Bacterial Zn(II)-Dependent Phospholipase C through Consensus Sequence Design.
Val, Diego S; Di Nardo, Luisina; Marchisio, Fiorela; Peiru, Salvador; Castelli, María Eugenia; Abriata, Luciano Andrés; Menzella, Hugo G; Rasia, Rodolfo M.
Afiliação
  • Val DS; Instituto de Procesos Biotecnológicos y Químicos (IPROBYQ), FbioyF-UNR-CONICET. Mitre 1998, 2000 Rosario, Argentina.
  • Di Nardo L; Instituto de Biología Celular y Molecular de Rosario (IBR), FbioyF-UNR-CONICET. Ocampo y Esmeralda, 2000 Rosario, Argentina.
  • Marchisio F; Instituto de Procesos Biotecnológicos y Químicos (IPROBYQ), FbioyF-UNR-CONICET. Mitre 1998, 2000 Rosario, Argentina.
  • Peiru S; Keclon S.A., Cerrito 847, 2000 Rosario, Argentina.
  • Castelli ME; Instituto de Procesos Biotecnológicos y Químicos (IPROBYQ), FbioyF-UNR-CONICET. Mitre 1998, 2000 Rosario, Argentina.
  • Abriata LA; Ecole Polytechnique Fédérale de Lausanne, CH-1015 Lausanne, Switzerland.
  • Menzella HG; Instituto de Procesos Biotecnológicos y Químicos (IPROBYQ), FbioyF-UNR-CONICET. Mitre 1998, 2000 Rosario, Argentina.
  • Rasia RM; Instituto de Biología Celular y Molecular de Rosario (IBR), FbioyF-UNR-CONICET. Ocampo y Esmeralda, 2000 Rosario, Argentina.
Biochemistry ; 63(3): 348-354, 2024 02 06.
Article em En | MEDLINE | ID: mdl-38206322
ABSTRACT
Proteins' extraordinary performance in recognition and catalysis has led to their use in a range of applications. However, proteins obtained from natural sources are oftentimes not suitable for direct use in industrial or diagnostic setups. Natural proteins, evolved to optimally perform a task in physiological conditions, usually lack the stability required to be used in harsher conditions. Therefore, the alteration of the stability of proteins is commonly pursued in protein engineering studies. Here, we achieved a substantial thermal stabilization of a bacterial Zn(II)-dependent phospholipase C by consensus sequence design. We retrieved and analyzed sequenced homologues from different sources, selecting a subset of examples for expression and characterization. A non-natural consensus sequence showed the highest stability and activity among those tested. Comparison of the stability parameters of this stabilized mutant and other natural variants bearing similar mutations allows us to pinpoint the sites most likely to be responsible for the enhancement. Point mutations in these sites alter the unfolding process of the consensus sequence. We show that the stabilized version of the protein retains full activity even in harsh oil degumming conditions, making it suitable for industrial applications.
Assuntos
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Zinco / Proteínas Idioma: En Revista: Biochemistry Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Argentina País de publicação: Estados Unidos
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Zinco / Proteínas Idioma: En Revista: Biochemistry Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Argentina País de publicação: Estados Unidos