Expression of Glucose gene from Aspergilllus niger within pYES2/CT vector in Saccharomyces cereviceae inVSc1 for Glucose Biosensor Application.

Abstract

EXPRESSIONOFGLUCOSEOXIDASEGENEFROM ASPERGILLUSNIGERWITHINpYES2/ CT VECTOR IN SACCHAROMYCES CEREVISIAE INVSc1 FOR GLUCOSE BIOSENSOR APPLICATION. Glucose oxidase (GOX) enzyme has been applied as a raw material of glucose biosensor. GOX enzyme catalyses the oxidation of β-D-glucose into D-glucono-δ-lactone and hydrogen peroxide (H2O2) using oxygen as an electron acceptor. The purpose of this study was to express recombinant GOX gene originated from Indonesia’s Aspergillus niger strain in Saccharomyces cerevisiae INVSc1. Recombinant plasmid pYES2/CT containing GOX gene was successfully transformed into S. cerevisiae INVSc1 using the LiAc method. The total proteins induced by of 2, 4 and 8 % galactose inducer were extracted using various protocolswithglass beadslysesmethodastheoptimalextractiontechnique.Then,theproteinswereanalyzed foritsactivityusingglucoseliquidassay.Subsequently,theproteinswereusedasmaterialforglucosebiosensor, and the biosensor was tested for its electrochemical potential. The result showed that the recombinant GOX protein induced with 8 % galactose for 48 hours had a higher activity than commercial GOX. The Scanning ElectronMicroscopyanalysisshowedthattherecombinantGOXproteinwasdistributedmorehomogeneously on biosensor strip than that of commercial GOX. In the future, this recombinant GOX protein could be used asmaterialofglucosebiosensor.