gtme 构建共发酵木糖和葡萄糖的重组酿酒酵母 - semantic scholar.pdf

生 物 工 程 学 报 Chin J Biotech 2008, June 25; 24(6): 1010-1015 Chinese Journal of Biotechnology ISSN 1000-3061 cjb@ © 2008 Institute of Microbiology, CAS CSM, All rights reserved 研究报告 gTME 构建共发酵木糖和葡萄糖的重组酿酒酵母 刘红梅, 许琳, 严明, 来灿钢, 欧阳平凯 南京工业大学制药与生命科学学院, 材料化学工程国家重点实验室, 南京 210009 摘 要: 利用全转录工程（gTME ）方法将全局转录因子spt15 随机突变并克隆表达, 构建突变库。将突变基因连接到 表达载体 pYX212 上, 醋酸锂法转化入不利用木糖的酿酒酵母 YPH499 中, 经特定的培养基初筛获得高效利用木糖并 共发酵木糖和葡萄糖的酿酒酵母重组菌株。对获得的重组菌株进行了初步研究, 该菌株能够很好的利用木糖并共发酵木 糖和葡萄糖。在 30oC, 200 r/min, 发酵 96 h 时, 50 g/L 木糖和葡萄糖的利用率为94.0%和98.9%, 乙醇产率为32.4%和 31.6%, 原始菌株乙醇产率为 44.3%; 当木糖和葡萄糖以质量比 1:1 混合发酵时, 木糖和葡萄糖利用率分别为 91.7%和 85.9%, 乙醇产率为26% 。木糖醇的含量极低。 关键词: 全转录工程（gTME ）, 重组酵母菌, 木糖, 共发酵 gTME for Construction of Recombinant Yeast Co-fermenting Xylose and Glucose Hongmei Liu, Lin Xu, Ming Yan, Cangang Lai, and Pingkai Ouyang State Key Laboratory of Materials-Oriented Chemical Engineering, College of Life Science and Pharmaceutical Engineering, Nanjing University of Technology, Nanjing 210009, China Abstract: Global transcription machinery engineering (gTME) was employed to engineer xylose metabolism. Mutation of the transcription factor gene Spt15 was introduced by error-prone PCR, followed by screening on media using xylose as the sole carbon source. One recombinant strain growing well on such media was chosen for further research. This strain showed modest growth rates in the media containing 50 g/L xylose or glucose at the condition of 30oC, 200 r/min, 96 h, 94.0% and 98.9% of xylose and glucose were consumed, with the ethanol yield were 32.4% and 31.6%, respectively. The co