1. Systematic engineering for production of anti-aging sunscreen compound in Pseudomonas putida.
- Author
-
Yunus, Ian S., Hudson, Graham A., Chen, Yan, Gin, Jennifer W., Kim, Joonhoon, Baidoo, Edward E.K., Petzold, Christopher J., Adams, Paul D., Simmons, Blake A., Mukhopadhyay, Aindrila, Keasling, Jay D., and Lee, Taek Soon
- Subjects
- *
PSEUDOMONAS putida , *SUNSCREENS (Cosmetics) , *AGING prevention , *BINDING sites , *SYNTHETIC biology , *METABOLIC models - Abstract
Sunscreen has been used for thousands of years to protect skin from ultraviolet radiation. However, the use of modern commercial sunscreen containing oxybenzone, ZnO, and TiO 2 has raised concerns due to their negative effects on human health and the environment. In this study, we aim to establish an efficient microbial platform for production of shinorine, a UV light absorbing compound with anti-aging properties. First, we methodically selected an appropriate host for shinorine production by analyzing central carbon flux distribution data from prior studies alongside predictions from genome-scale metabolic models (GEMs). We enhanced shinorine productivity through CRISPRi-mediated downregulation and utilized shotgun proteomics to pinpoint potential competing pathways. Simultaneously, we improved the shinorine biosynthetic pathway by refining its design, optimizing promoter usage, and altering the strength of ribosome binding sites. Finally, we conducted amino acid feeding experiments under various conditions to identify the key limiting factors in shinorine production. The study combines meta-analysis of 13C-metabolic flux analysis, GEMs, synthetic biology, CRISPRi-mediated gene downregulation, and omics analysis to improve shinorine production, demonstrating the potential of Pseudomonas putida KT2440 as platform for shinorine production. • Meta-Analysis of 13C-MFA and GEMs show P. putida to be an ideal host for shinorine production. • CRISPRi-mediated gene downregulation of PP_1444 improves shinorine production. • Shinorine was produced at high titers and amino acid supplementation improved the titer. • Shinorine accumulates exclusively in the supernatant in P. putida cultures. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF