Construction of recombinant sestc Saccharomyces cerevisiae for consolidated bioprocessing, cellulase characterization, and ethanol production by in situ fermentation.

Bioethanol is an important oil substitute produced by the sugar fermentation process. To improve the efficiency of cellulase expression of Saccharomyces cerevisiae, a eukaryotic expression vector harboring a single-enzyme-system-three-cellulase gene (sestc) was integrated into the S. cerevisiae genome by the protoplast method. Using PCR screening, RT-PCR, and "transparent circle" detection, several recombinant S. cerevisiae strains, capable of efficiently expressing the heterogeneous cellulase, were selected. The total activity of cellulase, endo-β-D-glucanase, exo-β-D-glucanase, and xylanase of the recombinant S. cerevisiae transformant (designated number 14) was 1.1, 378, 1.44, and 164 U ml, respectively, which was 27.5-, 63-, 24-, and 19-fold higher than that of the wild-type strain. The concentration of ethanol produced by the engineered S. cerevisiae strain was 8.1 gl, with wheat bran as the carbon source, under submerged conditions; this was 57.86-fold higher than that produced by the wild-type strain (0.14 gl).