gTME for Improved Adaptation of Saccharomyces cerevisiae to Corn Cob Acid Hydrolysate
gTME for Improved Adaptation of Saccharomyces cerevisiae to Corn Cob Acid Hydrolysate
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gTME 改善酿酒酵母对玉米芯酸水解产物的适应
DOI：
10.1007/s12010-011-9201-7
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发表时间：
2011-08-01
影响因子：
3
通讯作者：
Ouyang, Pingkai
中科院分区：
文献类型：
--
作者：
Liu, Hongmei;Liu, Kai;Ouyang, Pingkai
利用全域转录机器工程技术对木质纤维素水解产物的木糖代谢、耐受性和适应性进行了研究。采用易错PCR法对转录因子spt 15进行突变，将重组质粒pYX 212-spt 15转化到不能在木糖上生长的酿酒酵母YPH 499中，并在以木质纤维素水解物为唯一碳源的培养基上进行筛选。最大糖产率通过用3%HCl（m/v）使用在120°C下高压灭菌2小时以1：10的固液比水解形式获得。玉米芯水解液含有68.41g/L木糖和7.67g/L葡萄糖。重组菌在不同预处理和pH条件的纤维素水解液中培养时，表现出适度的生长速率。结果表明，在初始pH为5的酸性水解液中，菌株spt 15 -29发酵71 h后，木糖和葡萄糖的利用率分别达到最大值65.7%和87.0%，最大乙醇浓度为11.9 g/L。
Global transcription machinery engineering was employed to engineer xylose metabolism, tolerance and adaptation to lignocellulosic hydrolysates. Mutation of the transcription factor spt15 was introduced by error-prone PCR, then transformed the recombinant plasmid pYX212-spt15 into Saccharomyces cerevisiae YPH499 which was not able to grow on xylose, and screened on media using lignocellulosic hydrolysates as the sole carbon source. The maximum sugar yield was obtained by the hydrolysis form with 3% HCl (m/v) using autoclaving at 120°C, for 2 h with a solid to liquid ratio of 1:10. The corn cob hydrolysate contained 68.41 g/L xylose and 7.67 g/L glucose. The recombinant strain showed modest growth rate when cultured in the cellulosic hyrolysates with different pretreatment and pH conditions. The results showed that spt15-29 reached the maximum xylose and glucose utilization of 65.7% and 87.0%, respectively, the maximum ethanol concentration was 11.9 g/L, after 71 h, using the acid hydrolysate with the initial pH of 5.