Abstract
The depolymerization of xylo-oligosaccharides to xylose by β-xylosidase and the subsequent ethanol production from xylose are important for reducing inhibition towards cellulase and hemicellulase and for taking full advantage of all sugars in lignocellulose hydrolysate. The activity of β-xylosidase in most cellulases produced by filamentous fungi is usually deficient, and thus recombinant xylose-fermenting Saccharomyces cerevisiae capable of secreting highly active β-xylosidase is increasingly recognized as an excellent candidate for addressing this issue. To this end, a prominent chassis cell, BSPX042, constructed in our previous work with a well-modified downstream metabolic pathway of xylose through multiple chromosome manipulations and beneficial mutations after evolution engineering, was selected as the host. The xylose isomerase coding gene Ru-xylA (where Ru represents the rumen), which was screened from the metagenomics library of bovine rumen contents, was shown to exhibit a high activity level in S. cerevisiae, and the glycoside hydrolase family 3 β-xylosidase gene xyl3A from Penicillium oxalicum was coexpressed in BSPX042. The recombinant strain BSGIBX with the highest extracellular activity of β-xylosidase was determined by comparing different signal peptides. The hydrolysis of xylobiose and xylotriose by BSGIBX culture further confirmed its β-xylosidase activity. BSGIBX can grow and produce ethanol with xylo-oligosaccharides as the sole carbon source. When xylo-oligosaccharides were pretreated by effective xylanase, the majority of the xylo-oligosaccharides were consumed rapidly and produced ~ 19.4 g L−1 ethanol at 48 h with a yield of 0.473 g g−1 consumed sugars in mixture with glucose. These results indicate the successful coexpression of β-xylosidase and xylose isomerase in ethanol-producing S. cerevisiae, and provide theoretical support for our future work in robust industrial yeast strains.
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Acknowledgments
This work was supported by the National Key R&D Program of China (2018YFB1501401, 2018YFB1501702), the National Natural Science Foundation of China (31870063), the Key R&D Project of Shandong Province (2017CXGC1105), the Major Program of Shandong Province Natural Science Foundation (ZR2018ZB0209) and the China Postdoctoral Science Foundation (2019M652374).
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Yaping Niu and Longhao Wu have contributed equally to this work and should be considered as co-first authors.
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Fig. S1
The fermentation chromatograms of strain BSGIBX in SC-X. (PPTX 56 kb)
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Niu, Y., Wu, L., Shen, Y. et al. Coexpression of β-xylosidase and xylose isomerase in Saccharomyces cerevisiae improves the efficiency of saccharification and fermentation from xylo-oligosaccharides. Cellulose 26, 7923–7937 (2019). https://doi.org/10.1007/s10570-019-02650-3
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DOI: https://doi.org/10.1007/s10570-019-02650-3