Breeding of a xylose-fermenting hybrid strain by mating genetically engineered haploid strains derived from industrial Saccharomyces cerevisiae
Breeding of a xylose-fermenting hybrid strain by mating genetically engineered haploid strains derived from industrial Saccharomyces cerevisiae
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DOI：
10.1007/s10295-014-1531-3
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发表时间：
2014-12-01
影响因子：
3.4
通讯作者：
Sawayama, Shigeki
中科院分区：
文献类型：
--
作者：
Inoue, Hiroyuki;Hashimoto, Seitaro;Sawayama, Shigeki
工业酿酒酵母IR-2是利用木质纤维素水解液发酵生产乙醇的一株很有前途的木糖利用酵母。基于木酮糖发酵速率选择两种基于IR-2的单倍体菌株，并通过交配含有异源木糖脱氢酶（XDH）（野生型NAD（+）依赖性XDH或工程化NADP（+）依赖性XDH，ARSdR）、木糖还原酶（XR）和木酮糖激酶（XK）基因的重组单倍体菌株获得杂种。在酵母提取物-蛋白胨-葡萄糖（YPD）琼脂和YP-木糖琼脂平板上选择生长速率的杂交体中，ARSdR的活性通常高于NAD（+）依赖性XDH。此外，具有相同水平的异质XDH活性的杂交菌株SE 12的木糖发酵性能与携带单组基因XR/ARSdR/XK的IR-2的重组菌株的木糖发酵性能相似。这些结果表明，重组单倍体菌株不仅保留了适当的遗传背景IR-2的乙醇生产从木糖，但也ARSdR是优选的木糖发酵。
The industrial Saccharomyces cerevisiae IR-2 is a promising host strain to genetically engineer xylose-utilizing yeasts for ethanol fermentation from lignocellulosic hydrolysates. Two IR-2-based haploid strains were selected based upon the rate of xylulose fermentation, and hybrids were obtained by mating recombinant haploid strains harboring heterogeneous xylose dehydrogenase (XDH) (wild-type NAD(+)-dependent XDH or engineered NADP(+)-dependent XDH, ARSdR), xylose reductase (XR) and xylulose kinase (XK) genes. ARSdR in the hybrids selected for growth rates on yeast extract-peptone-dextrose (YPD) agar and YP-xylose agar plates typically had a higher activity than NAD(+)-dependent XDH. Furthermore, the xylose-fermenting performance of the hybrid strain SE12 with the same level of heterogeneous XDH activity was similar to that of a recombinant strain of IR-2 harboring a single set of genes, XR/ARSdR/XK. These results suggest not only that the recombinant haploid strains retain the appropriate genetic background of IR-2 for ethanol production from xylose but also that ARSdR is preferable for xylose fermentation.