Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan.
Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan; Engineering Biology Research Center, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan.
Metab Eng. 2019 Dec;56:17-27. doi: 10.1016/j.ymben.2019.08.012. Epub 2019 Aug 18.
1,2,4-Butanetriol can be used to produce energetic plasticizer as well as several pharmaceutical compounds. Although Saccharomyces cerevisiae has some attractive characters such as high robustness for industrial production of useful chemicals by fermentation, 1,2,4-butanetriol production by S. cerevisiae has not been reported. 1,2,4-butanteriotl is produced by an oxidative xylose metabolic pathway completely different from the xylose reductase-xylitol dehydrogenase and the xylose isomerase pathways conventionally used for xylose assimilation in S. cerevisiae. In the present study, S. cerevisiae was engineered to produce 1,2,4-butanetriol by overexpression of xylose dehydrogenase (XylB), xylonate dehydratase (XylD), and 2-ketoacid decarboxylase. Further improvement of the recombinant strain was performed by the screening of optimal 2-ketoacid decarboxylase suitable for 1,2,4-butanetriol production and the enhancement of Fe uptake ability to improve the XylD enzymatic activity. Eventually, 1.7 g/L of 1,2,4-butanetriol was produced from 10 g/L xylose with a molar yield of 24.5%. Furthermore, 1.1 g/L of 1,2,4-butanetriol was successfully produced by direct fermentation of rice straw hydrolysate.
1,2,4-丁三醇可用作生产高能增塑剂以及几种医药化合物。尽管酿酒酵母具有一些吸引人的特点,如在发酵生产有用化学品方面具有较高的工业生产稳健性,但尚未有报道称酿酒酵母可以生产 1,2,4-丁三醇。1,2,4-丁三醇是通过一种完全不同于酿酒酵母中传统用于木糖同化的木糖还原酶-木糖醇脱氢酶和木糖异构酶途径的氧化木糖代谢途径产生的。在本研究中,通过过表达木酮糖脱氢酶(XylB)、木酮酸盐脱水酶(XylD)和 2-酮酸脱羧酶,对酿酒酵母进行了工程改造以生产 1,2,4-丁三醇。通过筛选适合 1,2,4-丁三醇生产的最佳 2-酮酸脱羧酶,并增强铁摄取能力以提高 XylD 酶活性,进一步改进了重组菌株。最终,从 10g/L 木糖中生产出了 1.7g/L 的 1,2,4-丁三醇,摩尔收率为 24.5%。此外,通过直接发酵稻草水解物成功生产出了 1.1g/L 的 1,2,4-丁三醇。