Total enzymatic synthesis of cis-α-irone from a simple carbon source

• Published in: Nature communications Vol. 13; no. 1; pp. 7421 - 10
• Main Authors: Chen, Xixian, T, Rehka, Esque, Jérémy, Zhang, Congqiang, Shukal, Sudha, Lim, Chin Chin, Ong, Leonard, Smith, Derek, André, Isabelle
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 02.12.2022; Nature Publishing Group; Nature Portfolio
• Subjects: 140/58; 631/45/603; 631/61/318; 639/638/92/607/1172; 82/1; 82/80; 82/83; Biochemistry, Molecular Biology; Bioreactors; Carbon; Carbon sources; Chemical Sciences; Design; Engineering; Enzymatic synthesis; Enzymes; Humanities and Social Sciences; Life Sciences; Metabolic engineering; Methyltransferase; Methyltransferases; Microorganisms; multidisciplinary; Natural products; Norisoprenoids; Protein Biosynthesis; Science; Science (multidisciplinary); Metabolic engineering; Transferases; Biocatalysis
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-022-35232-2

Metabolic engineering has become an attractive method for the efficient production of natural products. However, one important pre-requisite is to establish the biosynthetic pathways. Many commercially interesting molecules cannot be biosynthesized as their native biochemical pathways are not fully elucidated. Cis-α-irone, a top-end perfumery molecule, is an example. Retrobiosynthetic pathway design by employing promiscuous enzymes provides an alternative solution to this challenge. In this work, we design a synthetic pathway to produce cis-α-irone with a promiscuous methyltransferase (pMT). Using structure-guided enzyme engineering strategies, we improve pMT activity and specificity towards cis-α-irone by >10,000-fold and >1000-fold, respectively. By incorporating the optimized methyltransferase into our engineered microbial cells, ~86 mg l −1 cis-α-irone is produced from glucose in a 5 l bioreactor. Our work illustrates that integrated retrobiosynthetic pathway design and enzyme engineering can offer opportunities to expand the scope of natural molecules that can be biosynthesized. Retrosynthetic pathway design using promiscuous enzymes can provide a solution to the biosynthetic production of natural products. Here, the authors design a pathway for the production of cis-α-irone with a promiscuous methyltransferase using structure-guided enzyme engineering strategies.