β-Cryptoxanthin Production in Escherichia coli by Optimization of the Cytochrome P450 CYP97H1 Activity

• DOI: 10.1021/acs.jafc.2c08970
• Publication Date: 2023-03-08T19:42:25Z
• Authors: Thomas Lautier; Derek J. Smith; Lay Kien Yang; Xixian Chen; Congqiang Zhang; Gilles Truan; Nic D. Lindley
• Keywords: Heterologous; Metabolic Engineering; Heterologous expression; Synthetic Biology; Hydroxylation

Cytochromes P450, forming a superfamily of monooxygenases containing heme as cofactor, show great versatility in substrate specificity. Metabolic engineering can take advantage this feature to unlock novel metabolic pathways. However, the cytochromes P450 often difficulty being expressed heterologous chassis. As case study prokaryotic host Escherichia coli, synthesis β-cryptoxanthin was addressed. This carotenoid intermediate is difficult produce, its requires monoterminal hydroxylation β-carotene whereas most classic carotene hydroxylases are dihydroxylases. focused on optimization vivo activity CYP97H1, an original monohydroxylase. Engineering N-terminal part identifying matching redox partners, defining optimal cellular background and adjusting culture induction conditions improved production by 400 times compared that initial strain, representing 2.7 mg/L 20% total carotenoids produced.