Key Laboratory of Microbiology of Heilongjiang Province, College of Life Science, Heilongjiang University, Harbin 150080, People's Republic of China.
J Ind Microbiol Biotechnol. 2012 May;39(5):777-87. doi: 10.1007/s10295-011-1076-7. Epub 2012 Jan 24.
Genome shuffling is an efficient approach for the rapid improvement of industrially important microbial phenotypes. This report describes optimized conditions for protoplast preparation, regeneration, inactivation, and fusion using the Saccharomyces cerevisiae W5 strain. Ethanol production was confirmed by TTC (triphenyl tetrazolium chloride) screening and high-performance liquid chromatography (HPLC). A genetically stable, high ethanol-producing strain that fermented xylose and glucose was obtained following three rounds of genome shuffling. After fermentation for 84 h, the high ethanol-producing S. cerevisiae GS3-10 strain (which utilized 69.48 and 100% of the xylose and glucose stores, respectively) produced 26.65 g/L ethanol, i.e., 47.08% higher than ethanol production by S. cerevisiae W5 (18.12 g/L). The utilization ratios of xylose and glucose were 69.48 and 100%, compared to 14.83 and 100% for W5, respectively. The ethanol yield was 0.40 g/g (ethanol/consumed glucose and xylose), i.e., 17.65% higher than the yield by S. cerevisiae W5 (0.34 g/g).
基因组改组是快速改善工业重要微生物表型的有效方法。本报告描述了使用酿酒酵母 W5 菌株进行原生质体制备、再生、失活和融合的优化条件。通过 TTC（三苯基氯化四氮唑）筛选和高效液相色谱（HPLC）确认了乙醇的生产。经过三轮基因组改组，获得了一株遗传稳定、高产乙醇的菌株，该菌株能够发酵木糖和葡萄糖。经过 84 h 的发酵，高乙醇生产的 S. cerevisiae GS3-10 菌株（分别利用了木糖和葡萄糖库的 69.48%和 100%）生产了 26.65 g/L 的乙醇，比 S. cerevisiae W5（18.12 g/L）的乙醇产量高 47.08%。木糖和葡萄糖的利用率分别为 69.48%和 100%，而 W5 分别为 14.83%和 100%。乙醇得率为 0.40 g/g（乙醇/消耗的葡萄糖和木糖），比 S. cerevisiae W5 的得率 0.34 g/g 高 17.65%。
