Microbial biosynthesis of the anticoagulant precursor 4-hydroxycoumarin

• Published in: Nature communications Vol. 4; no. 1; p. 2603
• Main Authors: Lin, Yuheng, Shen, Xiaolin, Yuan, Qipeng, Yan, Yajun
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 16-10-2013; Nature Publishing Group
• Subjects: 4-Hydroxycoumarins - biosynthesis; 631/61/252/318; 692/699/75/593/1839; Anticoagulants - metabolism; Bacterial Proteins - biosynthesis; Bacterial Proteins - genetics; Bacterial Proteins - isolation & purification; Escherichia coli - enzymology; Escherichia coli - genetics; Humanities and Social Sciences; Humans; Kinetics; Ligases - biosynthesis; Ligases - genetics; Ligases - isolation & purification; Metabolic Engineering; Metabolic Networks and Pathways; multidisciplinary; Prodrugs - metabolism; Pseudomonas aeruginosa - enzymology; Pseudomonas aeruginosa - genetics; Science; Science (multidisciplinary); Warfarin - metabolism
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms3603

4-Hydroxycoumarin (4HC) type anticoagulants (for example, warfarin) are known to have a significant role in the treatment of thromboembolic diseases—a leading cause of patient morbidity and mortality worldwide. 4HC serves as an immediate precursor of these synthetic anticoagulants. Although 4HC was initially identified as a naturally occurring product, its biosynthesis has not been fully elucidated. Here we present the design, validation,
in vitro
diagnosis and optimization of an artificial biosynthetic mechanism leading to the microbial biosynthesis of 4HC. Remarkably, function-based enzyme bioprospecting leads to the identification of a characteristic FabH-like quinolone synthase from
Pseudomonas aeruginosa
with high efficiency on the 4HC-forming reaction, which promotes the high-level
de novo
biosynthesis of 4HC in
Escherichia coli
(~500 mg l
−1
in shake flasks) and further
in situ
semisynthesis of warfarin. This work has the potential to be scaled-up for microbial production of 4HC and opens up the possibility of biosynthesizing diverse coumarin molecules with pharmaceutical importance.
4-hydroxycoumarin (4HC), a precursor for anticoagulant drugs such as warfarin, has a major role in the treatment of thromboembolic diseases. Here, the authors present an artificial biosynthetic pathway for 4HC production in
E. coli
and demonstrate its potential for large-scale microbial production.