Abstract
Lacking the system balance of Aspergillus niger cellulase limits synergistic saccharification of biomass material. To improve the overall expression level of A. niger cellulase system, eukaryotic expression vector containing Ampullaria gigas spix cellulase gene was constructed. Using the method of protoplast-mediated transformation, cellulase gene from A. gigas spix was genetically integrated into A. niger genome. The enoki mushroom glyceraldehyde-3-phosphate dehydrogenase (gpd) promoter could effectively drive gene expression of exogenous cellulase sestc gene in A.niger. Filter paper activity (total cellulase activity) of the transformant No. 17 (1.736 ± 0.051 Uml−1) induced by wheat bran was 1.21-fold higher compared with that of the A.niger wild type. Beta-endo-1-4-glucanase, beta-exo-1-4-glucanase, and xylanase produced by the engineered A.niger were 1.37-fold, 1.25-fold, and 1.3-fold higher than those of the wild-type strain. Total cellulase activity of A. niger transformant No. 17 induced by alkaline-pretreated rice straw reached 1.476 ± 0.021 FPU ml−1, which was 1.31-fold higher compared with the wild type strain.
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