A Selective Fluorescent l-Lactate Biosensor Based on an l-Lactate-Specific Transcription Regulator and Forster Resonance Energy Transfer

Selective detection of l-lactate levels in foods, clinical, and bacterial fermentation samples has drawn intensive attention. Many fluorescent biosensors based on non-stereoselective recognition elements have been developed for lactate detection. Herein, the allosteric transcription factor STLldR from Salmonella enterica serovar Typhimurium LT2 was identified to be stereo-selectively respond to l-lactate. Then, STLldR was combined with Forster resonance energy transfer (FRET) to construct a fluorescent l-lactate biosensor FILLac. FILLac was further optimized by truncating the N- and C-terminal amino acids of STLldR between cyan and yellow fluorescent proteins. The optimized biosensor FILLac(10N0C) exhibited a maximum emission ratio change (Delta R-max) of 33.47 +/- 1.91%, an apparent dissociation constant (K-d) of 6.33 +/- 0.79 mu M, and a limit of detection of 0.68 mu M. FILLac(10N0C) was applied in 96-well microplates to detect l-lactate in bacterial fermentation samples and commercial foods such as Jiaosu and yogurt. The quantitation results of FILLac(10N0C) exhibited good agreement with that of a commercial l-lactate biosensor SBA-40D bioanalyzer. Thus, the biosensor FILLac(10N0C) compatible with high-throughput detection may be a potential choice for quantitation of l-lactate in different biological samples.

Automated summary

In this study, a fluorescent biosensor FILLac(10N0C) was developed for the selective detection of L-lactate levels. It showed high sensitivity and a low limit of detection, making it suitable for high-throughput detection in various biological samples.