Light-driven progesterone production by InP-(M. neoaurum) biohybrid system

Abstract Progesterone is one of the classical hormone drugs used in medicine for maintaining pregnancy. However, its manufacturing process, coupled with organic reagents and poisonous catalysts, results in irreversible environmental pollution. Recent advances in synthetic biology have demonstrated the microbial biosynthesis of natural products, especially difficult-to-synthesize compounds, synthetic from building blocks is a promising strategy. Herein, overcoming the heterologous cytochrome P450 enzyme interdependency in Mycolicibacterium neoaurum successfully constructed the CYP11A1 running module to realize the metabolic conversion from waste phytosterols to progesterone. Subsequently, progesterone yield was improved through strategies involving electron transfer and NADPH regeneration. The CYP11A1 and ADR were connected by a flexible linker as a chimera to enhance electron transfer. Strain MNR-08 showed positive activity, with 45 mg/L progesterone, leading to a 3.95-fold improvement over strain MNR-04. Significantly, a novel inorganic-biological hybrid system was assembled by combining MNR-08 and InP nanoparticles to regenerate NADPH, which was increased 84-fold from the initial progesterone titers to 235 ± 50 mg/L. Altogether, this work highlights the green and sustainable potential for synthetic progesterone from sterols in M. neoaurum.