Deflavination of Bacillus sp. Sarcosine Oxidase and Reconstitution of Flavoproteins

Abstract:

Due to the unsatisfactory effects of deflavination from sarcosine oxidase（SOX），the SOX inclusion protein without cofactor was dialysed and renatured for testing the reconstruction effects between SOX apoprotein and the coenzymes. The comparatively suitable conditions for the binding of flavoprotein to SOX apoprotein were shown as following：the concentration of enzyme protein 0.3 g/L， pH 8.5， temperature 4 ℃， the optimal redox state（GSH / GSSG ） GSH / GSSG with the value 2，and the performing time was 24 h. The reconstitution effects of SOX protein binding to different natural flavins were compared. Through CD，FTIR，and DSC analysis，the secondary structure，enzyme specific activity and stability of the reconstituted SOX were also studied.