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作 者:孙锦云[1] 高文萱[1] 丁文涛[1] 刘文[1] 陈洵[1]
机构地区:[1]天津大学化工学院,天津300072
出 处:《化学工业与工程》2015年第1期64-70,78,共8页Chemical Industry and Engineering
摘 要:以双拷贝过表达木糖代谢上游途径关键酶(木糖还原酶XR、木糖醇脱氢酶XDH和木酮糖激酶XKS)的酿酒酵母菌株为背景,在过表达非氧化磷酸戊糖(PP)途径中转醛酶基因TAL1的基础上,对途径中其他基因TKL1(转酮酶)、RPE1(核酮糖-5-磷酸差向异构酶)和RKI1(核酮糖-5-磷酸异构酶)进行了不同程度的过表达,以研究PP途径基因过表达对酿酒酵母木糖代谢的影响。在不同培养基条件下对重组菌株木糖代谢进行研究,结果显示,在过表达TAL1的基础上不同组合过表达PP途径其他基因不同程度改善了酿酒酵母木糖发酵性能,重组菌株能在36-48 h耗完质量分数(下同)为5%的木糖。其中,过表达PP途径全部基因比其他过表达基因组合表现出明显的优势,在8%木糖发酵条件下其乙醇产量达到了每1 g木糖0.337 g,较对照菌株提高了7.86%。这说明同步过表达PP途径基因更有利于酿酒酵母木糖发酵。To study the influence of non-oxidative pentose phosphate pathway (PPP) on xylose utilization in Saccharomyces cerevisiae, different combinations of PPP genes TKL1, RPE1 and RKI1 were overexpressed in the TALl-enhanced yeast strain, which also had the double-copy integration of upstream path- way genes XYL1, XYL2 and XKS1. Performance of recombinant yeasts was investigated at both 5 % and 8 % xylose as well as glucose and xylose mixture, results showed that overproduction of PPP genes stimu- lated cell growth and xylose conversion at both xylose concentrations, concomitant with higher ethanol yield at the expense of xylitol and glycerol formation, and all recombinant strains can use up 5 % xylose within 36 to 48 h. Yeast strains of S. cerevisiae with four PPP genes co-overexpressed showed more advantages, and under the 8 % xylose concentration, ethanol yield was about 0. 337 g/g xylose, 7.86 % higher than parent strain. It indicated that co-overproduction of all PPP genes may be the top priority for enhancing xylose metabolism in S. cerevislae strains.
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