Tuning architectural organization of eukaryotic P450 system to boost bioproduction in Escherichia coli.

Autor: Li Y; Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, 210014, China.; Jiangsu Key Laboratory for the Research and Utilization of Plant Resources, Nanjing, 210014, China.; Nanjing Botanical Garden Mem. Sun Yat-Sen, Nanjing, 210014, China., Li J; Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, 210014, China.; Jiangsu Key Laboratory for the Research and Utilization of Plant Resources, Nanjing, 210014, China.; Nanjing Botanical Garden Mem. Sun Yat-Sen, Nanjing, 210014, China., Chen WK; Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, 210014, China., Li Y; Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, 210014, China., Xu S; Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, 210014, China.; Jiangsu Key Laboratory for the Research and Utilization of Plant Resources, Nanjing, 210014, China.; Nanjing Botanical Garden Mem. Sun Yat-Sen, Nanjing, 210014, China., Li L; Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, 210014, China., Xia B; Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, 210014, China.; Jiangsu Key Laboratory for the Research and Utilization of Plant Resources, Nanjing, 210014, China.; Nanjing Botanical Garden Mem. Sun Yat-Sen, Nanjing, 210014, China., Wang R; Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nanjing, 210014, China. rwang@cnbg.net.; Jiangsu Key Laboratory for the Research and Utilization of Plant Resources, Nanjing, 210014, China. rwang@cnbg.net.; Nanjing Botanical Garden Mem. Sun Yat-Sen, Nanjing, 210014, China. rwang@cnbg.net.
Jazyk: angličtina
Zdroj: Nature communications [Nat Commun] 2024 Nov 19; Vol. 15 (1), pp. 10009. Date of Electronic Publication: 2024 Nov 19.
DOI: 10.1038/s41467-024-54259-1
Abstrakt: Eukaryotic cytochrome P450 enzymes, generally colocalizing with their redox partner cytochrome P450 reductase (CPR) on the cytoplasmic surface of organelle membranes, often perform poorly in prokaryotic cells, whether expressed with CPR as a tandem chimera or free-floating individuals, causing a low titer of heterologous chemicals. To improve their biosynthetic performance in Escherichia coli, here, we architecturally design self-assembled alternatives of eukaryotic P450 system using reconstructed P450 and CPR, and create a set of N-termini-bridged P450-CPR heterodimers as the counterparts of eukaryotic P450 system with N-terminus-guided colocalization. The covalent counterparts show superior and robust biosynthetic performance, and the N-termini-bridged architecture is validated to improve the biosynthetic performance of both plant and human P450 systems. Furthermore, the architectural configuration of protein assemblies has an inherent effect on the biosynthetic performance of N-termini-bridged P450-CPR heterodimers. The results suggest that spatial architecture-guided protein assembly could serve as an efficient strategy for improving the biosynthetic performance of protein complexes, particularly those related to eukaryotic membranes, in prokaryotic and even eukaryotic hosts.
Competing Interests: Competing interests The authors declare no competing interests.
(© 2024. The Author(s).)
Databáze: MEDLINE
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