Biosynthetic bifunctional enzyme complex with high-efficiency luciferin-recycling to enhance the bioluminescence imaging

Hill, Philip; Sun, Xiaohui; Tang, Xu; Hu, Rui; Luo, Man; HILL, PHIL; Fang, Baishan; Xu, Chang'an

Abstract

Firefly luciferase is a prominent reporter on molecular imaging with the advantage of longer wavelength on light emission and the ATP linear correlation, which makes it useful in most of current bioluminescence imaging model. However, the utility of this biomaterial was limited by the signal intensity and stability which are respectively affected by enzyme activity and substrate consumption. This study demonstrated a series of novel synthetic bifunctional enzyme complex of Firefly luciferase (Fluc) and Luciferin-regenerating enzyme (LRE). A peptide linker library was constructed for the fusion strategy on biosynthesis. The findings of both experimental data and structural simulation demonstrated that the intervention of fused LRE remarkably improve the stability of in vitro bioluminescence signal through luciferin recycling; and revealed the competitive relationship of Fluc and LRE on luciferin binding: Fluc performed higher activity with one copy number of rigid linker (EAAAK) at the C terminal while LRE acted more efficiently with two copy numbers of flexible linker (GGGGS) at the N terminal. With the advantage of signal intensity and stability, this fused bifunctional enzyme complex may expand the application of firefly luciferase to in vitro bioluminescence imaging.

Citation

Hill, P., Sun, X., Tang, X., Hu, R., Luo, M., HILL, P., Fang, B., & Xu, C. (2019). Biosynthetic bifunctional enzyme complex with high-efficiency luciferin-recycling to enhance the bioluminescence imaging. International Journal of Biological Macromolecules, 130, 705-714. https://doi.org/10.1016/j.ijbiomac.2019.03.036

• Journal Article Type: Article
• Acceptance Date: Mar 4, 2019
• Online Publication Date: Mar 5, 2019
• Publication Date: Jun 1, 2019
• Deposit Date: Mar 11, 2019
• Publicly Available Date: Mar 6, 2020
• Journal: International Journal of Biological Macromolecules
• Print ISSN: 0141-8130
• Electronic ISSN: 1879-0003
• Publisher: Elsevier
• Peer Reviewed: Peer Reviewed
• Volume: 130
• Pages: 705-714
• DOI: https://doi.org/10.1016/j.ijbiomac.2019.03.036
• Keywords: Biochemistry; Molecular Biology; Structural Biology; General Medicine
• Public URL: https://nottingham-repository.worktribe.com/output/1624028
• Publisher URL: https://www.sciencedirect.com/science/article/pii/S0141813019303976?via%3Dihub
• Additional Information: This article is maintained by: Elsevier; Article Title: Biosynthetic bifunctional enzyme complex with high-efficiency luciferin-recycling to enhance the bioluminescence imaging; Journal Title: International Journal of Biological Macromolecules; CrossRef DOI link to publisher maintained version: https://doi.org/10.1016/j.ijbiomac.2019.03.036; Content Type: article; Copyright: © 2019 Elsevier B.V. All rights reserved.
• Contract Date: Mar 11, 2019