Toyota Biotechnology & Afforestation Laboratory, Toyota Motor Co, 1099 Marune, Kurozasa-cho, Miyoshi, Aichi, 470-0201, Japan.
Appl Microbiol Biotechnol. 2011 Sep;91(6):1553-9. doi: 10.1007/s00253-011-3357-5. Epub 2011 Jun 4.
We constructed a recombinant industrial Saccharomyces cerevisiae yeast strain OC2-AXYL2-ABGL2-Xyl2 by inserting two copies of the β-glucosidase (BGL) and β-xylosidase (XYL) genes, and a gene cassette for xylose assimilation in the genome of yeast strain OC-2HUT. Both BGL and XYL were expressed on the yeast cell surface with high enzyme activities. Using OC2-AXYL2-ABGL2-Xyl2, we performed ethanol fermentation from a mixture of powdered cellulose (KC-flock) and Birchwood xylan, with the additional supplementation of a 30-g/l Trichoderma reesei cellulase complex mixture. The ethanol yield (gram per gram of added cellulases) of the strain OC2-AXYL2-ABGL2-Xyl2 increased approximately 2.5-fold compared to that of strain OC2-Xyl2, which lacked β-glucosidase and β-xylosidase activities. Notably, the concentration of additional T. reesei cellulase was reduced from 30 to 24 g/l without affecting ethanol production. The BGL- and XYL-displaying industrial yeast of the strain OC2-AXYL2-ABGL2-Xyl2 represents a promising yeast for reducing cellulase consumption of ethanol fermentation from lignocellulosic biomass by compensating for the inherent weak BGL and XYL activities of T. reesei cellulase complexes.
我们通过在酵母菌株 OC-2HUT 的基因组中插入两个β-葡萄糖苷酶(BGL)和β-木糖苷酶(XYL)基因以及一个木糖同化基因盒,构建了重组工业酿酒酵母菌株 OC2-AXYL2-ABGL2-Xyl2。BGL 和 XYL 均在酵母细胞表面表达,具有较高的酶活。使用 OC2-AXYL2-ABGL2-Xyl2,我们在添加了 30 g/L 里氏木霉纤维素酶复合混合物的条件下,从粉状纤维素(KC- flock)和桦木木聚糖的混合物中进行乙醇发酵。与缺乏β-葡萄糖苷酶和β-木糖苷酶活性的菌株 OC2-Xyl2 相比,菌株 OC2-AXYL2-ABGL2-Xyl2 的乙醇产率(每克添加纤维素酶的克数)提高了约 2.5 倍。值得注意的是,在不影响乙醇生产的情况下,里氏木霉纤维素酶的添加量从 30 g/L 降低到 24 g/L。该菌株 OC2-AXYL2-ABGL2-Xyl2 中展示 BGL 和 XYL 的工业酵母代表了一种很有前途的酵母,可通过补偿里氏木霉纤维素酶复合物固有的较弱的 BGL 和 XYL 活性来降低乙醇发酵中纤维素酶的消耗,从而利用木质纤维素生物质进行生产。