A Single Mutation in the Carbohydrate-Binding Module Enhances Cellulase Activity in Bacillus Amyloliquefaciens Mutant

From our earlier work, we modified the carbohydrate-binding module (CBM) of Bacillus amyloliquefaciens to increase cellulase activity using cold plasma technology. The gene (BglC-M) from mutant was expressed in Escherichia coli BL21(DE3) under T7 promoter. hydrolysis approximately 2.5-fold higher than control (BglC-W) over a wide range pH and temperature conditions. amino acid sequence BglC-M contained 471 residues that were almost identical BglC-W. Only single acid, lysine, replaced by glutamic at position 370 (K370E) within (CBM). Structure prediction substrate docking indicated mutation might involve cellulose binding β-sandwich facilitated hydrogen bonding. study cellopentaose with model structure replacement lysine-370 led formation bond 436Y, which has shorter distance (2.6 Å) compared (5.4 Å). As result, becomes more compact stable, resulting increased catalytic efficiency. Finally, biomass ability investigated on lignocellulosic wastes such as pineapple peel, corncob, durian peel. enzyme showed significant amount reducing sugar released all control. This first evidence altering base composition enhanced activity.
 HIGHLIGHTS
 
 Increasing technology
 Mutation cellulose-binding enhance activity
 Greater wastes
 GRAPHICAL ABSTRACT