Caffeic acid production from glucose using metabolically engineered Escherichia coli

• Published in: Enzyme and microbial technology 2023-03, Vol.164, p.110193-110193, Article 110193
• Main Authors: Sakae, Kosuke, Nonaka, Daisuke, Kishida, Mayumi, Hirata, Yuuki, Fujiwara, Ryosuke, Kondo, Akihiko, Noda, Shuhei, Tanaka, Tsutomu
• Format: Article
• Language: English
• Subjects: Caffeic acid; Caffeic Acids - metabolism; E. coli; Escherichia coli - genetics; Escherichia coli - metabolism; Glucose; Metabolic Engineering; Mixed Function Oxygenases; p-coumaric acid; Shikimate pathway
• ISSN: 0141-0229; 1879-0909
• DOI: 10.1016/j.enzmictec.2023.110193

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. [Display omitted] •E. coli expressing both of SeC3H and PaHpaB/PaHpaC monooxygenases was constructed.•FAD regeneration by PaHpaC significantly improved caffeic acid production.•An engineered strain directly produced 6.17 g/L of caffeic acid from glucose.