VII Simposio Internacional de Química 2019 "SIQ 2019" -XII Conferencia "La Ingeniería Química: Desarrollo, potencialidades y sus retos"

VII Simposio Internacional de Química 2019

SIQ 2019

RECOMBINANT PRODUCTION OF ENZYMES FOR USE IN THE SUGAR AND FOOD INDUSTRIES

Resumen [ES]

Abstract We will present our group results in the recombinant production of two enzymes (fructosyltransferase and invertase) that convert sucrose into value-added products and one enzyme (dextranase) used as aid to sugar production and refining. Constructed plasmids carrying several in-tandem copies of the transgene fused to a secretory signal peptide and under the transcriptional control of the constitutive glyceraldehyde 3-phosphate dehydrogenase (GAP) promoter were integrated in the Pichia pastoris genome. After fed-batch fermentations, each recombinant enzyme was recovered with purity above 85% from culture supernatants and characterized in terms of yield, posttranslational modifications, biochemical properties and catalytic performance. Fructosyltransferase converted sucrose to short-chain fructooligosaccharides (scFOS), particularly the most relevant prebiotic 1-kestose. Invertase completely hydrolyzed sucrose at elevated temperatures (60-70°C). The endo-type dextranase degraded microbial dextran and did not react on sucrose. For commercial purposes, each enzyme was recovered from the yeast culture supernatant and freeze-dried to a stable soluble powder. The recombinant P. pastoris strains developed in this study are cost-effective sources for the scaled production of three enzymes with application in the sugar and/or food industry.

Resumen [EN]

Abstract We will present our group results in the recombinant production of two enzymes (fructosyltransferase and invertase) that convert sucrose into value-added products and one enzyme (dextranase) used as aid to sugar production and refining. Constructed plasmids carrying several in-tandem copies of the transgene fused to a secretory signal peptide and under the transcriptional control of the constitutive glyceraldehyde 3-phosphate dehydrogenase (GAP) promoter were integrated in the Pichia pastoris genome. After fed-batch fermentations, each recombinant enzyme was recovered with purity above 85% from culture supernatants and characterized in terms of yield, posttranslational modifications, biochemical properties and catalytic performance. Fructosyltransferase converted sucrose to short-chain fructooligosaccharides (scFOS), particularly the most relevant prebiotic 1-kestose. Invertase completely hydrolyzed sucrose at elevated temperatures (60-70°C). The endo-type dextranase degraded microbial dextran and did not react on sucrose. For commercial purposes, each enzyme was recovered from the yeast culture supernatant and freeze-dried to a stable soluble powder. The recombinant P. pastoris strains developed in this study are cost-effective sources for the scaled production of three enzymes with application in the sugar and/or food industry.

Sobre el ponente

Carmen Menéndez Rodríguez

Dr. Carmen Menéndez Rodríguez

CIGB Habana Flag of Cuba
Información Práctica
Ponencia
Spanish / Español
No definido
30 minutos
No definido
Autores
Eulogio Pimentel
Duniesky Martínez
Dr. Carmen Menéndez Rodríguez
Enrique R. Pérez
Osmani Chacón
Alina Sobrino
Yamira Quintero
Ricardo Ramírez
Lázaro Hernández
Ana G. Martínez
Palabras clave
dextranase
fructooligosaccharide
fructosyltransferase
invert syrup
invertase
pichia pastoris