Conversion of Mevalonate to Isoprenol Using Light Energy in Escherichia coli without Consuming Sugars for ATP Supply

• Published in: ACS synthetic biology 2022-12, Vol.11 (12), p.3966-3972
• Main Authors: Sano, Mikoto, Tanaka, Ryo, Kamata, Kentaro, Hirono-Hara, Yoko, Ishii, Jun, Matsuda, Fumio, Hara, Kiyotaka Y., Shimizu, Hiroshi, Toya, Yoshihiro
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphate - metabolism; Escherichia coli - genetics; Escherichia coli - metabolism; Mevalonic Acid - metabolism; Rhodopsin - genetics; Rhodopsin - metabolism; Sugars - metabolism
• ISSN: 2161-5063; 2161-5063
• DOI: 10.1021/acssynbio.2c00313

Bioconversion of key intermediate metabolites such as mevalonate into various useful chemicals is a promising strategy for microbial production. However, the conversion of mevalonate into isoprenoids requires a supply of adenosine triphosphate (ATP). Light-driven ATP regeneration using microbial rhodopsin is an attractive module for improving the intracellular ATP supply. In the present study, we demonstrated the ATP-consuming conversion of mevalonate to isoprenol using rhodopsin-expressing Escherichia coli cells as a whole-cell catalyst in a medium that does not contain energy cosubstrate, such as glucose. Heterologous genes for the synthesis of isoprenol from mevalonate, which requires three ATP molecules for the series of reactions, and a delta-rhodopsin gene derived from Haloterrigena turkmenica were cointroduced into E. coli. To evaluate the conversion efficiency of mevalonate to isoprenol, the cells were suspended in a synthetic medium containing mevalonate as the sole carbon source and incubated under dark or light illumination (100 μmol m–2 s–1). The specific isoprenol production rates were 10.0 ± 0.9 and 20.4 ± 0.7 μmol gDCW–1 h–1 for dark and light conditions, respectively. The conversion was successfully enhanced under the light condition. Furthermore, the conversion efficiency increased with increasing illumination intensity, suggesting that ATP regenerated by the proton motive force generated by rhodopsin using light energy can drive ATP-consuming reactions in the whole-cell catalyst.