Construction and optimization of 1, 4-butanediamine biosensor based on transcriptional regulator PuuR.

Junjie LIU; Minmin JIANG; Tong SUN; Xiangxiang SUN; Yongcan ZHAO; Mingxia GU; Fuping LU; Ming LI

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Construction and optimization of 1, 4-butanediamine biosensor based on transcriptional regulator PuuR.

Author: Junjie LIU ¹ ; Minmin JIANG ¹ ; Tong SUN ¹ ; Xiangxiang SUN ¹ ; Yongcan ZHAO ¹ ; Mingxia GU ¹ ; Fuping LU ¹ ; Ming LI ¹
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1. Key Laboratory of Industrial Fermentation Microbiology (Tianjin University of Science & Technology), Ministry of Education, Tianjin 300457, China.
Publication Type:Journal Article
Keywords: 1,4-butanediamine; Escherichia coli; biosensor; transcriptional regulator PuuR
MeSH: Biosensing Techniques/methods*; Escherichia coli/metabolism*; Promoter Regions, Genetic/genetics*; Green Fluorescent Proteins/metabolism*; Transcription Factors/genetics*; Escherichia coli Proteins/genetics*; Diamines/metabolism*; Gene Expression Regulation, Bacterial
From: Chinese Journal of Biotechnology 2025;41(1):437-447
CountryChina
Language:Chinese
Abstract: Biosensors have become powerful tools for real-time monitoring of specific small molecules and precise control of gene expression in biological systems. High-throughput sensors for 1, 4-butanediamine biosynthesis can greatly improve the screening efficiency of high-yielding 1, 4-butanediamine strains. However, the strategies for adapting the characteristics of biosensors are still rarely studied, which limits the applicability of 1, 4-butanediamine biosensors. In this paper, we propose the development of a 1, 4-butanediamine biosensor based on the transcriptional regulator PuuR, whose homologous operator puuO is installed in the constitutive promoter P_gapA of Escherichia coli to control the expression of the downstream superfolder green fluorescent protein (sfGFP) as the reporter protein. Finally, the biosensor showed a stable linear relationship between the GFP/OD₆₀₀ value and the concentration of 1, 4-butanediamine when the concentration of 1, 4-butanediamine was 0-50 mmol/L. The promoters with different strengths in the E. coli genome were used to modify the 1, 4-butanediamine biosensor, and the functional properties of the PuuR-based 1, 4-butanediamine biosensor were explored and improved, which laid the groundwork for high-throughput screening of engineered strains highly producing 1, 4-butanediamine.