期刊
BIOSENSORS & BIOELECTRONICS
卷 211, 期 -, 页码 -出版社
ELSEVIER ADVANCED TECHNOLOGY
DOI: 10.1016/j.bios.2022.114378
关键词
Lactate; Allosteric transcription factor; LldR; Biosensor; Amplified luminescent proximity homogeneous; assay
类别
资金
- National Key R&D Program of China [2019YFA0904800, 2019YFA0904900]
- National Natural Science Foundation of China [31970056]
- State Key Laboratory of Microbial Technology Open Projects Fund [M2021-12]
- Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project [TSBICIP-KJGG-005]
A biosensor capable of detecting both D-lactate and L-lactate was developed in this study, using an allosteric transcription factor LldR and amplified luminescent proximity homogeneous assay technology. The optimized biosensor, BLac-6, showed high accuracy in lactate concentration detection and was tentatively applied in industrial fermentation.
Lactate, a hydroxycarboxylic acid commercially produced by microbial fermentation, is widely applied in diverse industrial fields. Lactate exists in two stereoisomeric forms (D-lactate and L-lactate). D-Lactate and L-lactate are often simultaneously present in many biological samples. Therefore, a biosensor able to detect both D- and Llactate is required but previously unavailable. Herein, an allosteric transcription factor LldR from Pseudomonas aeruginosa PAO1, which responds to both D-lactate and L-lactate, was combined with amplified luminescent proximity homogeneous assay technology to develop a D,L-lactate biosensor. The proposed biosensor was optimized by mutation of DNA sequence in binding site of LldR. The optimized biosensor BLac-6 can accurately detect the concentration of lactate independent on ratio of the two isomers in pending test samples. The biosensor was also tentatively used in quantitative analysis of D-lactate, L-lactate, or D,L-lactate in fermentation samples produced by three recombinant strains of Klebsiella oxytoca. With its desirable properties, the biosensor BLac-6 may be a potential choice for monitoring the concentration of lactate during industrial fermentation.
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