Upgraded cellulose and xylan digestions for synergistic enhancements of biomass enzymatic saccharification and bioethanol conversion using engineered Trichoderma reesei strains overproducing mushroom LeGH7 enzyme.

Crop straws provide enormous lignocellulose resources transformable for sustainable biofuels and valuable bioproducts. However, lignocellulose recalcitrance basically restricts essential biomass enzymatic saccharification at large scale. In this study, the mushroom-derived cellobiohydrolase (LeGH7) was introduced into Trichoderma reesei (Rut-C30) to generate two desirable strains, namely GH7-5 and GH7-6. Compared to the Rut-C30 strain, both engineered strains exhibited significantly enhanced enzymatic activities, with ß-glucosidases, endocellulases, cellobiohydrolases, and xylanase activities increasing by 113 %, 140 %, 241 %, and 196 %, respectively. By performing steam explosion and mild alkali pretreatments with mature straws of five bioenergy crops, diverse lignocellulose substrates were effectively digested by the crude enzymes secreted from the engineered strains, leading to the high-yield hexoses released for bioethanol production. Notably, the LeGH7 enzyme purified from engineered strain enabled to act as multiple cellulases and xylanase at higher activities, interpreting how synergistic enhancement of enzymatic saccharification was achieved for distinct lignocellulose substrates in major bioenergy crops. Therefore, this study has identified a novel enzyme that is active for simultaneous hydrolyses of cellulose and xylan, providing an applicable strategy for high biomass enzymatic saccharification and bioethanol conversion in bioenergy crops.

Cellulose de-polymerization is selective for bioethanol refinery and multi-functional biochar assembly using brittle stalk of corn mutant.

As lignocellulose recalcitrance principally restricts for a cost-effective conversion into biofuels and bioproducts, this study re-selected the brittle stalk of corn mutant by MuDR-transposon insertion, and detected much reduced cellulose polymerization and crystallinity. Using recyclable CaO chemical for biomass pretreatment, we determined a consistently enhanced enzymatic saccharification of pretreated corn brittle stalk for higher-yield bioethanol conversion. Furthermore, the enzyme-undigestible lignocellulose was treated with two-step thermal-chemical processes via FeCl2 catalysis and KOH activation to generate the biochar with significantly raised adsorption capacities with two industry dyes (methylene blue and Congo red). However, the desirable biochar was attained from one-step KOH treatment with the entire brittle stalk, which was characterized as the highly-porous nanocarbon that is of the largest specific surface area at 1697.34 m2/g and 2-fold higher dyes adsorption. Notably, this nanocarbon enabled to eliminate the most toxic compounds released from CaO pretreatment and enzymatic hydrolysis, and also showed much improved electrochemical performance with specific capacitance at 205 F/g. Hence, this work has raised a mechanism model to interpret how the recalcitrance-reduced lignocellulose is convertible for high-yield bioethanol and multiple-function biochar with high performance.

A mechanism for efficient cadmium phytoremediation and high bioethanol production by combined mild chemical pretreatments with desirable rapeseed stalks.

Cadmium (Cd) is one of the most hazardous trace metals, and rapeseed is a major oil crop over the world with considerable lignocellulose residues applicable for trace metal phytoremediation and cellulosic ethanol co-production. In this study, we examined that two distinct rapeseed cultivars could accumulate Cd at 72.48 and 43.70 ug/g dry stalk, being the highest Cd accumulation among all major agricultural food crops as previously reported. The Cd accumulation significantly increased pectin deposition as a major factor for trace metal association with lignocellulose. Meanwhile, the Cd-accumulated rapeseed stalks contained much reduced wall polymers (hemicellulose, lignin) and cellulose degree of polymerization, leading to improved lignocellulose enzymatic hydrolysis. Notably, three optimal chemical pretreatments were performed for enhanced biomass enzymatic saccharification and bioethanol production by significantly increasing cellulose accessibility and lignocellulose porosity, along with a complete Cd release for collection and recycling. Hence, this study proposed a mechanism model interpreting why rapeseed stalks are able to accumulate much Cd and how the Cd-accumulated stalks are of enhanced biomass saccharification. It has also provided a powerful technology for both cost-effective Cd phytoremediation and value-added bioethanol co-production with minimum waste release.