Sakae Kosuke, Nonaka Daisuke, Kishida Mayumi, Hirata Yuuki, Fujiwara Ryosuke, Kondo Akihiko, Noda Shuhei, Tanaka Tsutomu
Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
Center for Sustainable Resource Science, RIKEN, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan; Graduate School of Science, Technology and Innovation, Kobe University, 1-1 Rokkodai, Nada, Kobe 657-8501, Japan.
Enzyme Microb Technol. 2023 Mar;164:110193. doi: 10.1016/j.enzmictec.2023.110193. Epub 2023 Jan 5.
Caffeic acid (3,4-dihydroxycinnamic acid) is a precursor for high-valued compounds with anticancer, antiviral activities, and anti-inflammatory making it an important substance in the food additive, cosmetics, and pharmaceutical industries. Here, we developed an engineered Escherichia coli strain capable of directly producing high levels of caffeic acid from glucose. Tyrosine ammonia-lyase from Rhodotorula glutinis (RgTAL) and p-coumaric acid 3-hydroxylase from Saccharothrix espanaensis (SeC3H) were expressed. Next, feedback-resistant chorismate mutase/prephenate dehydrogenase, was introduced to promote l-tyrosine synthesis. This engineered strain CA3 produced 1.58 g/L of caffeic acid from glucose without tyrosine supplemented to the medium. Furthermore, to reduce p-coumaric acid accumulation, 4-hydroxyphenylacetate 3-hydroxylase from Pseudomonas aeruginosa (PaHpaBC) was introduced. Finally, an engineered strain CA8 directly produced 6.17 g/L of caffeic acid from glucose using a jar fermenter. The E. coli developed in this study would be helpful as a chassis strain to produce value-added caffeic acid-derivatives.
咖啡酸（3,4-二羟基肉桂酸）是具有抗癌、抗病毒活性和抗炎作用的高价值化合物的前体，这使其成为食品添加剂、化妆品和制药行业中的重要物质。在此，我们构建了一种能够直接从葡萄糖高产咖啡酸的工程化大肠杆菌菌株。表达了来自粘红酵母的酪氨酸解氨酶（RgTAL）和来自西班牙糖丝菌的对香豆酸3-羟化酶（SeC3H）。接下来，引入了对反馈抑制不敏感的分支酸变位酶/预苯酸脱氢酶，以促进L-酪氨酸的合成。该工程菌株CA3在培养基中不添加酪氨酸的情况下，从葡萄糖中产生了1.58 g/L的咖啡酸。此外，为了减少对香豆酸的积累，引入了来自铜绿假单胞菌的4-羟基苯乙酸3-羟化酶（PaHpaBC）。最后，工程菌株CA8使用罐式发酵罐从葡萄糖中直接产生了6.17 g/L的咖啡酸。本研究构建的大肠杆菌作为底盘菌株有助于生产增值的咖啡酸衍生物。
