Optimization of the culture medium composition using response surface methodology for new recombinant cyprosin B production in bioreactor for cheese production

The optimization of culture medium compositionwas carried out for improvement the recombinantcyprosin B production, an enzyme with high milk-clottingactivity. Response surface methodology (RSM) wasapplied to evaluate the effect of variables namely glucose,yeast extract (YE) and bactopeptone present...

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Bibliographic Details
Main Author: Sampaio, Pedro N. (author)
Other Authors: Calado, Cecília R. C. (author), Sousa, Lisete (author), Bressler, David C. (author), Pais, Maria Salomé (author), Fonseca, Luís Pina (author)
Format: article
Language:eng
Published: 2011
Subjects:
Online Access:http://hdl.handle.net/10400.14/6422
Country:Portugal
Oai:oai:repositorio.ucp.pt:10400.14/6422
Description
Summary:The optimization of culture medium compositionwas carried out for improvement the recombinantcyprosin B production, an enzyme with high milk-clottingactivity. Response surface methodology (RSM) wasapplied to evaluate the effect of variables namely glucose,yeast extract (YE) and bactopeptone present in the culturemedium, used for recombinant cyprosin B production bytransformed Saccharomyces cerevisiae BJ1991 strain inshake-flask and bioreactor culture conditions. The centracomposite experimental design (CCD) was adopted toderive a statistical model for optimizing the composition ofthe fermentation medium. The optimal concentration estimatedfor each variable related to a theoretical maximumof cyprosin B activity (488 U mL-1) was 30 g L-1 glucose,15 g L-1 YE and 27 g L-1 bactopeptone. The optimizedmedium composition, based on empirical model, ledto a cyprosin B activity of 519 U mL-1, which correspondsto an increase of 46%. The fermentation using optimizedculture medium in a 5-L bioreactor allowed a significantincrease in biomass (82%) and recombinant cyprosin Bproduction (139%). The improvement in the recombinantcyprosin B production after optimization process can beconsidered adequate for large-scale applications, and theclotting activity of cyprosin B account for their use inindustrial cheese making.