Visualization of a Limonene Synthesis Metabolon Inside Living Bacteria by Hyperspectral SRS Microscopy. Issue 32 (28th September 2022)

• Record Type: Journal Article
• Authors: Zhang, Jing; Shin, Jonghyeon; Tague, Nathan; Lin, Haonan; Zhang, Meng; Ge, Xiaowei; Wong, Wilson; Dunlop, Mary J.; Cheng, Ji‐Xin
• Is Part Of: Advanced science. Volume 9:Issue 32(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-09-28
• Subjects: escherichia coli -- limonene -- metabolic engineering -- stimulated Raman scattering; Science -- Periodicals; 505
• DOI: 10.1002/advs.202203887
• Languages: English
• ISSNs: 2198-3844

Abstract

Abstract: Monitoring biosynthesis activity at single‐cell level is key to metabolic engineering but is still difficult to achieve in a label‐free manner. Using hyperspectral stimulated Raman scattering imaging in the 670–900 cm −1 region, localized limonene synthesis are visualized inside engineered Escherichia coli . The colocalization of limonene and GFP‐fused limonene synthase is confirmed by co‐registered stimulated Raman scattering and two‐photon fluorescence images. The finding suggests a limonene synthesis metabolon with a polar distribution inside the cells. This finding expands the knowledge of de novo limonene biosynthesis in engineered bacteria and highlights the potential of SRS chemical imaging in metabolic engineering research. Abstract : Monitoring biosynthesis activity at the single‐cell level is key to metabolic engineering but is still difficult to achieve in a label‐free manner. Here, the chemical composition and synthetic function of the limonene‐rich aggregates inside single engineered Escherichia coli is directly visualized using hyperspectral stimulated Raman scattering. It suggests a limonene synthesis metabolon inside the cells.