From Bugs to Bioplastics: Total (+)‐Dihydrocarvide Biosynthesis by Engineered Escherichia coli

• Published in: Chembiochem : a European journal of chemical biology Vol. 20; no. 6; pp. 785 - 792
• Main Authors: Ascue Avalos, Gabriel A., Toogood, Helen S., Tait, Shirley, Messiha, Hanan L., Scrutton, Nigel S.
• Format: Journal Article
• Language: English
• Published: Germany Wiley Subscription Services, Inc 15.03.2019; John Wiley and Sons Inc
• Subjects: (+)-dihydrocarvide monomer; Bacteria; Baeyer–Villiger monooxygenases; Biology; Bioplastics; Biosynthesis; Carbon sources; Carvone; Chassis; Chemical synthesis; Cyclohexanone; Cyclohexanone monooxygenase; E coli; engineering; Escherichia coli; Escherichia coli - enzymology; Escherichia coli - genetics; Escherichia coli - metabolism; Gene expression; Glucose - chemistry; Homology; Lactones - chemistry; Lactones - metabolism; Limonene; Metabolic Engineering - methods; Microorganisms; Monomers; Monoterpenes - chemistry; Monoterpenes - metabolism; Optimization; Organic chemistry; Polymers; Proof of Concept Study; Reductase; Screening; Shape memory; Stereoisomerism; Sustainable development; Synthetic biology; Synthetic Biology - methods; synthetic biology; Baeyer-Villiger monooxygenases; engineering; (+)-dihydrocarvide monomer; bioplastics
• ISSN: 1439-4227; 1439-7633
• DOI: 10.1002/cbic.201800606

The monoterpenoid lactone derivative (+)‐dihydrocarvide ((+)‐DHCD) can be polymerised to form shape‐memory polymers. Synthetic biology routes from simple, inexpensive carbon sources are an attractive, alternative route over chemical synthesis from (R)‐carvone. We have demonstrated a proof‐of‐principle in vivo approach for the complete biosynthesis of (+)‐DHCD from glucose in Escherichia coli (6.6 mg L−1). The pathway is based on the Mentha spicata route to (R)‐carvone, with the addition of an ′ene′‐reductase and Baeyer–Villiger cyclohexanone monooxygenase. Co‐expression with a limonene synthesis pathway enzyme enables complete biocatalytic production within one microbial chassis. (+)‐DHCD was successfully produced by screening multiple homologues of the pathway genes, combined with expression optimisation by selective promoter and/or ribosomal binding‐site screening. This study demonstrates the potential application of synthetic biology approaches in the development of truly sustainable and renewable bioplastic monomers. Sustainable, renewable bioplastic monomers: The monoterpenoid (+)‐ DHCD can be polymerised to form shape‐memory polymers. We have demonstrated a proof‐of‐principle approach to the complete biosynthesis of (+)‐ DHCD from glucose in E. coli, based on a modified M. spicata biosynthetic pathway. This route from a simple, inexpensive carbon source is more attractive than the alternative chemical synthesis from (R)‐carvone.