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

ABSTRACT: 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 Förster 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 (ΔR_max) of 33.47 ± 1.91%, an apparent dissociation constant (K_d) of 6.33 ± 0.79 μM, and a limit of detection of 0.68 μM. 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.

SUBMITTER: Xu X 

PROVIDER: S-EPMC9775004 | biostudies-literature | 2022 Dec

REPOSITORIES: biostudies-literature