Department of Bio and Fermentation Convergence Technology and BK21 Plus Program, Kookmin University, Seoul 02707, Korea.
Synthetic Biology and Bioengineering Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Korea.
Enzyme Microb Technol. 2017 Dec;107:7-14. doi: 10.1016/j.enzmictec.2017.07.014. Epub 2017 Jul 31.
Inefficient transport of xylose into Saccharomyces cerevisiae is a major hurdle for production of xylitol, a natural sweetener with five carbons. To facilitate the xylose transport and hence increase xylose conversion to xylitol, the araE gene encoding an arabinose:H symporter (AraE) from Bacillus subtilis and the XYL1 gene from Scheffersomyces stipitis were expressed in Saccharomyces cerevisiae EBY.VW4000, a hxt null mutant. The resulting strain of EXHA exhibited 4.1 fold increases in xylose consumption rate and xylitol productivity, relative to the control strain without AraE. Also, overexpression of AraE in wild type S. cerevisiae D452-2 having all hexose transporters and the XYL1 gene increased both xylose consumption and xylitol production considerably. In a glucose-limited fed-batch culture with intermittent addition of xylose, the DXXA strain with multiple copies of araE and XYL1 produced 177.8g/L xylitol with 2.47g/L-h productivity, which were 26.9 and 17.6 times higher than those for a batch culture of the DX strain expressing the XYL1 gene only, respectively. It was concluded that B. subtilis AraE might be a potent xylose transporter and conferred much higher xylose-consuming and xylitol-producing abilities to S. cerevisiae.
木糖向酵母中的低效运输是生产木糖醇(一种具有五个碳原子的天然甜味剂)的主要障碍。为了促进木糖的运输,从而提高木糖转化为木糖醇的效率,我们在酵母 EBY.VW4000(hxt 缺失突变体)中表达了来自枯草芽孢杆菌的阿拉伯糖:H 同向转运蛋白(AraE)的 araE 基因和来自酿酒酵母的 XYL1 基因。与没有 AraE 的对照菌株相比,表达 EXHA 的菌株的木糖消耗速率和木糖醇生产力分别提高了 4.1 倍。此外,在具有所有己糖转运蛋白和 XYL1 基因的野生型 S. cerevisiae D452-2 中过表达 AraE 也大大提高了木糖的消耗和木糖醇的生产。在间歇添加木糖的葡萄糖限制分批补料培养中,具有多个 araE 和 XYL1 拷贝的 DXXA 菌株生产了 177.8g/L 的木糖醇,生产率为 2.47g/L-h,分别比仅表达 XYL1 基因的 DX 菌株的分批培养高 26.9 倍和 17.6 倍。综上所述,枯草芽孢杆菌 AraE 可能是一种有效的木糖转运蛋白,赋予了酵母更高的木糖消耗和木糖醇生产能力。