Effective one-step saccharification of lignocellulosic biomass using magnetite-biocatalysts containing saccharifying enzymes.

Lignocellulosic biomass, packed with sugars, is one of the most available renewable resources for biofuels and bioproducts production. To release the sugars for the production, enzymatic hydrolysis (saccharification) of pretreated lignocellulosic biomass are required. However, the saccharification process is costly, inefficient, and requires multi-step operations. This is in part due to the high cost and the limited selection of commercial enzymes which commonly have different optimal pH and temperatures. Here we reported a one-step saccharification of pretreated lignocellulosic biomass using immobilized biocatalysts containing five different saccharifying enzymes (SEs) with a similar optimum pH and temperature. The five SEs - endo-1,4-β-d-glucanase (an endoglucanase, eglS), cellobiohydrolase (an exoglucanase, cbhA), and β-glucosidase (bglH), endo-1,4-β-xylanase (an endoxylanase, xynC) and β-xylosidase (bxlB) - were successfully expressed and produced by E. coli BL21. Better saccharification of pretreated corn husks was observed when using the five crude SE enzymes than those using two commonly used SEs, endo-1,4-β-d-glucanase and β-glucosidase. The five SEs were cross-linked in the absence or the presence of magnetic nanoparticles (hereafter referred as SE-CLEAs and M-SE-CLEAs, respectively). By using SE-CLEAs, the highest amount of reduced sugar (250 mg/g biomass) was measured. The activity of immobilized SEs is better than free crude SEs. The M-SE-CLEAs can be reused at least 3 times for effective saccharification of pretreated lignocellulosic biomass.