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In molecular docking, the frequencies of amino acid residue sites within 5 \u00c5 of CEL3A with ligand cellobiose, EG1 with ligand CMC, and CBH2 with ligand \u03b2-glucan are determined.Table S4. PCR primer of EG1,CBH2,CEL3A and its mutants.Table S5. The signal peptides from Saccharomyces cerevisiae which are used for the expression of exogenous proteins in Pichia pastoris and the intrinsic signal peptides of C. glycerinogenes.Fig. S1. Ramachandran plot assessment analysis of the modeling of CEL3A,EG1,CBH2. The (a), (c), (e) analyses were performed by PyMOL, the (b), (d), (f) were performed by AlphaFold 2Fig. S2. The protein concentration, total cellulase enzymatic activity, and specific enzymatic activity of the recombinant strains of the C.glycerinogenes complex cellulase after multi-strategy modification;Cg1 serves as the initial strain of the complex cellulase; Cg2 is the recombinant strain following signal peptide mining; Cg3 is the recombinant strain after semi-rational design modification of each cellulase; Cg4 is the recombinant strain integrating the signal peptide mining and semi-rational design strategyFig. S3. Determination of extracellular CEL3A-WT and Q91, R156 and D267 mutant residues activity; Determination of extracellular EG1-WT and T272 and C385 mutant residues activity; Determination of extracellular CBH-WT and V251, G289 and L294 mutant residues activityFig. S4. Superposition effect of CEL3A,EG1,CBH2. (a), (c), (e): Preliminary screening of mutants and superimposition schemes. (b), (d), (f): Catalytic activity of key mutants in the superposition processFig. S5. 10% SDS-PAGE analysis of composite cellulase and its engineered strain. Lane M, the molecular mass standard; Lanes 1 to 4, crude enzyme solution of wild composite cellulase; signal peptide-engineered composite cellulase; composite cellulase modified by site-directed mutagenesis; composite cellulase engineered based on signal peptide engineering and site-directed mutagenesis","order":4,"name":"Ethics","group":{"name":"EthicsHeading","label":"Support information"}}],"article-number":"68"}}