1. Engineering yeast with a light-driven proton pump system in the vacuolar membrane.
- Author
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Daicho, Kaoru M., Hirono-Hara, Yoko, Kikukawa, Hiroshi, Tamura, Kentaro, and Hara, Kiyotaka Y.
- Subjects
PROTONS ,RHODOPSIN ,SACCHAROMYCES cerevisiae ,BIOENGINEERING ,ENGINEERING - Abstract
Background: The supply of ATP is a limiting factor for cellular metabolism. Therefore, cell factories require a sufficient ATP supply to drive metabolism for efficient bioproduction. In the current study, a light-driven proton pump in the vacuolar membrane was constructed in yeast to reduce the ATP consumption required by V-ATPase to maintain the acidification of the vacuoles and increase the intracellular ATP supply for bioproduction. Results: Delta rhodopsin (dR), a microbial light-driven proton-pumping rhodopsin from Haloterrigena turkmenica, was expressed and localized in the vacuolar membrane of Saccharomyces cerevisiae by conjugation with a vacuolar membrane-localized protein. Vacuoles with dR were isolated from S. cerevisiae, and the light-driven proton pumping activity was evaluated based on the pH change outside the vacuoles. A light-induced increase in the intracellular ATP content was observed in yeast harboring vacuoles with dR. Conclusions: Yeast harboring the light-driven proton pump in the vacuolar membrane developed in this study are a potential optoenergetic cell factory suitable for various bioproduction applications. [ABSTRACT FROM AUTHOR]
- Published
- 2024
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