Development of a novel tailor-made cocktail from recombinant crude enzymes for efficient saccharification of...

A novel tailor-made enzyme cocktail comprising five recombinant crude enzymes was designed for hydrolysis of alkaline hydrogen peroxide pretreated elephant grass (AHPpEG). D-optimal design approach is employed to optimize the proportion of individual enzymes within the enzyme cocktail. The enzyme cocktail comprising chimera (endoglucanase and β-glucosidase), cellobiohydrolase, endo-1,4-β-xylanase, β-xylosidase and arabinofuranosidase having ratio (9:4:2:2:3) was developed. The crude enzyme cocktail was stable at 35 °C for 24 h, retaining over 60 % relative activity and displayed t_1/2 of 55 h. The saccharification of AHPpEG at 15 % (w/v) solid loading, the developed enzyme cocktail gave total reducing sugar yield of 465 mg/g_.AHPpEG. The addition of 2.0 g/L TritonX-100 to the crude enzyme cocktail for saccharification of 15 % (w/v) AHPpEG caused a significant enhancement (17 %) in TRS yield (544 mg/g_.AHPpEG) with glucose 435 mg/g_.AHPpEG and xylose 106 mg/g._AHPpEG yields. The formulated tailor-made crude enzyme cocktail for AHPpEG gave saccharification efficiency of 76 % underscoring the potential of the enzyme cocktail for application in lignocellulosic biomass based biorefineries.