Regulating ehrlich and demethiolation pathways for alcohols production by the expression of ubiquitin-protein ligase gene HUWE1.
Regulating ehrlich and demethiolation pathways for alcohols production by the expression of ubiquitin-protein ligase gene HUWE1.
复制标题
通过泛素蛋白连接酶基因 HUWE1 的表达调节乙醇生产的埃利希和去甲基硫基化途径
DOI：
10.1038/srep20828
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发表时间：
2016-02-10
影响因子：
4.6
通讯作者：
Tang YJ
中科院分区：
文献类型：
--
作者：
Zhang Q;Jia KZ;Xia ST;Xu YH;Liu RS;Li HM;Tang YJ
当两个相互竞争的分支将氨基酸转化为醇时，Ehrlich和去甲硫化途径。控制这两条途径为工业应用提供了相当大的潜力，包括酒精过量生产、风味质量控制和开发新口味。然而，如何规范Ehrlich和去甲基化途径仍不适用。以蛋氨酸转化为甲硫醇和甲硫醇为例，利用抑制消减杂交(SSH)技术构建了两个华支睾吸虫抑制消减cDNA文库，用于筛选控制甲硫醇转化的调控因子。从正向SSH文库中筛选到的E3泛素蛋白连接酶基因HUWE1被证实与末端产物的生物合成有关。HUWE1在玫瑰酵母和酿酒酵母中的过表达显著增加了甲硫醇及其衍生物在脱甲硫醇途径中的生物合成，而抑制了甲硫醇和甲硫醇在艾氏途径中的生物合成。这些结果通过HUWE1的过度表达实现了对两条通路的方向性调节。因此，HUWE1有可能成为通过代谢工程控制和提高酒精产量的关键靶点。
Ehrlich and demethiolation pathways as two competing branches converted amino acid into alcohols. Controlling both pathways offers considerable potential for industrial applications including alcohols overproduction, flavor-quality control and developing new flavors. While how to regulate ehrlich and demethiolation pathways is still not applicable. Taking the conversion of methionine into methionol and methanethiol for example, we constructed two suppression subtractive cDNA libraries of Clonostachys rosea by using suppression subtractive hybridization (SSH) technology for screening regulators controlling the conversion. E3 ubiquitin-protein ligase gene HUWE1 screened from forward SSH library was validated to be related with the biosynthesis of end products. Overexpressing HUWE1 in C. rosea and S. cerevisiae significantly increased the biosynthesis of methanethiol and its derivatives in demethiolation pathway, while suppressed the biosynthesis of methional and methionol in ehrlich pathway. These results attained the directional regulation of both pathways by overexpressing HUWE1. Thus, HUWE1 has potential to be a key target for controlling and enhancing alcohols production by metabolic engineering.