Characterization and synergistic activity of heterologously expressed microbial-derived endoglucanase and bifunctional cellulase on wheat straw.

Cellulases are divided into endoglucanase, exoglucanase, and β-glucosidase based on their catalytic activity. Eight cellulases were recombinantly expressed in Escherichia coli BL21 to investigate their effects on the enzymatic hydrolysis of wheat straw. Among them, cellulase 2006 exhibited the highest endoglucanase activity (432.25 U/mg), while Bf1 displayed superior exoglucanase and β-glucosidase activities (577.46 and 1991.63 U/mg respectively). Bioinformatic and enzymatic analyses revealed that both cellulases displayed notable thermal and pH stability. The enzyme kinetics parameters revealed that Km values for cellulases 2006 and Bf1 were 15.98 and 14.19 mg/mL, respectively, with Vmax values of 20.78 and 16.38 μmol/min/mg. In a prokaryotic co-expression system, the mixed cellulase Bf2006 exhibited endoglucanase, exoglucanase, and β-glucosidase activities (130.78, 1406.36, and 1119.25 U/mg, respectively). The enzymatic hydrolysis assay revealed that these three cellulases acted on the cellulose macromolecules in wheat straw, increasing reducing sugar content and decreasing crystallinity. Endoglucanase 2006 acted on various organic compounds rich in phenols and aromatic heterocycles, while Bf1 primarily acted on compounds containing glucose units. Bf2006 significantly affected the content of lignin, neutral detergent fiber, acid detergent fiber, and the microstructure of wheat straw, with degradation products primarily consisting of disaccharides, oligosaccharides, polysaccharides, glycosides, and other carbohydrates. This study provides theoretical guidance for the production and application of mixed cellulase Bf2006, serving as a reference for the industrial use of different cellulase types.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
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