Casein-based scaffold for artificial cellulosome design

Recombinant endoglucanase was prepared with the insertion of a microbial transglutaminase (MTG)-reactive lysine residue (K-tag) in its C-terminus. MTG-mediated conjugation was conducted to attach the recombinant endoglucanase to β-casein or Dimethylated casein. Two different cellulase clusters were observed after conjugation, and the impact of each distinct casein-EG conjugate was tested through enzymatic saccharification of cellulosic substrate, in order to analyze their potential as casein-based scaffold for cellulosome design. DM-casein-EG conjugates were more efficient than β-casein-EG-conjugates, with more than 2-fold increase when using the endoglucanase Cel5A.