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Background  It remains a challenge for recombinant  S. cerevisiae  to convert xylose in lignocellulosic biomass hydrolysates to ethanol. Although industrial diploid strains are more robust compared to laboratory haploid strains, however, industrial diploid  S. cerevisiae  strains have been less pursued in previous studies. This work aims to construct fast xylose-fermenting yeast using an industrial ethanol-producing diploid  S. cerevisiae  strain as a host.    Results  Fast xylose-fermenting yeast was constructed by genome integration of xylose-utilizing genes and adaptive evolution, including 1)  Piromyces XYLA  was introduced to enable the host strain to convert xylose to xylulose; 2) endogenous genes ( XKS1, RKI1, RPE1, TKL1 , and  TAL1 ) were overexpressed to accelerate conversion of xylulose to ethanol; 3)  Candida intermedia GXF1 , which encodes a xylose transporter, was introduced at the  GRE3  locus to improve xylose uptake; 4) aerobic evolution in rich xylose media was carried out to increase growth and xylose consumption rates. The best evolved strain CIBTS0735 consumed 80 g/l glucose and 40 g/l xylose in rich media within 24 hours at an initial OD 600  of 1.0 (0.63 g DCW/l) and produced 53 g/l ethanol.    Conclusions  Based on the above fermentation performance, we conclude that CIBTS0735 shows great potential for ethanol production from lignocellulosic biomass.

bmc biotechnology

Construction of fast xylose-fermenting yeast based on industrial ethanol-producing diploid Saccharomyces cerevisiaeby rational design and adaptive evolution