Shen Li, Nishimura Yuya, Matsuda Fumio, Ishii Jun, Kondo Akihiko
Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe 657-8501, Japan.
Organization of Advanced Science and Technology, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe, Hyogo 657-8501, Japan.
J Biosci Bioeng. 2016 Jul;122(1):34-9. doi: 10.1016/j.jbiosc.2015.12.022. Epub 2016 Mar 11.
2-Phenylethanol (2-PE) is a higher aromatic alcohol that is used in the cosmetics and food industries. The budding yeast Saccharomyces cerevisiae is considered to be a suitable host for the industrial production of higher alcohols, including 2-PE. To produce 2-PE from glucose in S. cerevisiae, we searched for suitable 2-keto acid decarboxylase (KDC) and alcohol dehydrogenase (ADH) enzymes of the Ehrlich pathway for overexpression in strain YPH499, and found that overexpression of the ARO10 and/or ADH1 genes increased 2-PE production from glucose. Further, we screened ten BY4741 single-deletion mutants of genes involved in the competing pathways for 2-PE production, and found that strains aro8Δ and aat2Δ displayed increased 2-PE production. Based on these results, we engineered a BY4741 strain that overexpressed ARO10 and contained an aro8Δ deletion, and demonstrated that the strain produced 96 mg/L 2-PE from glucose as the sole carbon source. As this engineered S. cerevisiae strain showed a significant increase in 2-PE production from glucose without the addition of an intermediate carbon substrate, it is a promising candidate for the large-scale production of 2-PE.
2-苯乙醇（2-PE）是一种高级芳香醇，用于化妆品和食品工业。酿酒酵母被认为是工业生产包括2-PE在内的高级醇的合适宿主。为了在酿酒酵母中由葡萄糖生产2-PE，我们寻找了埃利希途径中合适的2-酮酸脱羧酶（KDC）和醇脱氢酶（ADH）进行YPH499菌株中的过表达，发现ARO10和/或ADH1基因的过表达增加了由葡萄糖生产的2-PE。此外，我们筛选了参与2-PE生产竞争途径的基因的十个BY4741单缺失突变体，发现aro8Δ和aat2Δ菌株的2-PE产量增加。基于这些结果，我们构建了一个过表达ARO10且含有aro8Δ缺失的BY4741菌株，并证明该菌株以葡萄糖作为唯一碳源可产生96mg/L的2-PE。由于这种工程化的酿酒酵母菌株在不添加中间碳底物的情况下由葡萄糖生产2-PE的产量显著增加，它是大规模生产2-PE的有前途的候选菌株。
