title: Production of a modified peptide clavanin in Pichia pastoris: cloning, expression, purification and in vitro activities.
abstract: Antimicrobial peptides are one of the most promising peptide-based drugs due to their enormous potential as novel biopharmaceuticals in both human and animal industries. In order to develop strategies to over produce such molecules, heterologous production of a modified version of clavanin A, here named clavanin MO (clavMO), was successfully achieved in the methylothopic yeast Pichia pastoris. ClavMO was fused to thioredoxin as a carrier protein and the construction was tested using two promoters, PAOX1 and PGAP, based on either induced or constitutive expression systems, respectively. After growth in 5 L Bioreactor, clavMO-thio was recovered and purified through size exclusion chromatography. Our findings show that both constitutive and inducible expression systems produce active clavMO fused to thioredoxin against both Gram-negative Klebsiella pneumoniae and Gram-positive Staphylococcus aureus microorganisms.
authors.affiliation: Centro de Análises Proteômicas e Bioquímicas, Pós-Graduação em Ciências Genômicas e Biotecnologia, Universidade Católica de Brasília, Brasília, DF, Brazil, kellylmulder@gmail.com.
authors.forename: Kelly Cristina
authors.initials: KC
authors.lastname: Mulder
authors.forename: Loiane Alves
authors.initials: LA
authors.lastname: de Lima
authors.forename: Priscilla Santos
authors.initials: PS
authors.lastname: Aguiar
authors.forename: Fábio Correa
authors.initials: FC
authors.lastname: Carneiro
authors.forename: Octávio Luiz
authors.initials: OL
authors.lastname: Franco
authors.forename: Simoni Campos
authors.initials: SC
authors.lastname: Dias
authors.forename: Nádia Skorupa
authors.initials: NS
authors.lastname: Parachin