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为了获得耐发酵抑制剂、耐乙醇并利用木糖的纤维素乙醇生产菌种,以实验室保藏菌种Saccharomyces cerevisiaeY5和Pichia stipitis CBS6054为亲本,采用双亲灭活原生质体融合技术,选育出了共代谢葡萄糖和木糖,且耐受发酵抑制剂的酵母菌株Y10-F。该菌在以木糖为唯一碳源的培养基中培养96h,木糖的利用率达到58.8%,乙醇浓度为5.2g/L。在外加6.0、8.0g/L乙醇的培养液中,Y10-F的生长优于亲株。外加3.0g/L的糠醛时,Y10-F的延滞期较亲本Y5和CBS6054分别缩短了6h和18h。对融合菌株Y10-F进行了汽爆法预处理玉米秸秆的酶解液发酵实验,具有较好的耐毒和产乙醇能力。
In order to obtain the fermentation-resistant, ethanol-resistant and xylose-producing cellulosic ethanol-producing strains, parental inactivated Saccharomyces cerevisiae Y5 and Pichia stipitis CBS6054 were used as parents to inactivate protoplast fusion technique, Yeast strain Y10-F, which metabolizes glucose and xylose and is resistant to fermentation inhibitors. The strain was cultured for 96 h in a medium containing xylose as the sole carbon source. The xylose utilization rate was 58.8% and the ethanol concentration was 5.2 g / L. In the medium supplemented with 6.0 and 8.0 g / L ethanol, Y10-F grew better than the parent strain. With the addition of 3.0g / L furfural, the lag phase of Y10-F was shortened by 6h and 18h, respectively, than the parental Y5 and CBS6054. The fusion strain Y10-F was pre-treated by steam explosion method to hydrolyze corn stalk, which showed good resistance to toxicity and ethanol production.