|
KMID : 0545120230330040552
|
|
Journal of Microbiology and Biotechnology
2023 Volume.33 No. 4 p.552 ~ p.558
|
|
|
A Genetically Encoded Biosensor for the Detection of Levulinic Acid
|
|
Kim Jee-Hung
Woo Seung-Gyun
Kim Seong-Keun
Yoo Byeong-Hyeon
Shin Jong-Hyeok
Rha Eu-Gene
Kim Soo-Jung
Kwon Kil-Koang
Lee Hye-Won
Kim Ha-Seong
Kim Hee-Taek
Sung Bong-Hyun
Sohn Joo-Hyuk
Lee Dae-Hee
|
|
|
Abstract
|
|
|
|
Levulinic acid (LA) is a valuable chemical used in fuel additives, fragrances, and polymers. In this study, we proposed possible biosynthetic pathways for LA production from lignin and poly(ethylene terephthalate). We also created a genetically encoded biosensor responsive to LA, which can be used for screening and evolving the LA biosynthesis pathway genes, by employing an LvaR transcriptional regulator of Pseudomonas putida KT2440 to express a fluorescent reporter gene. The LvaR regulator senses LA as a cognate ligand. The LA biosensor was first examined in an Escherichia coli strain and was found to be non-functional. When the host of the LA biosensor was switched from E. coli to P. putida KT2440, the LA biosensor showed a linear correlation between fluorescence intensity and LA concentration in the range of 0.156?10 mM LA. In addition, we determined that 0.156 mM LA was the limit of LA detection in P. putida KT2440 harboring an LA-responsive biosensor. The maximal fluorescence increase was 12.3-fold in the presence of 10 mM LA compared to that in the absence of LA. The individual cell responses to LA concentrations reflected the population-averaged responses, which enabled high-throughput screening of enzymes and metabolic pathways involved in LA biosynthesis and sustainable production of LA in engineered microbes.
|
|
|
KEYWORD
|
|
|
Levulinic acid, genetically encoded biosensor, transcription factor, Pseudomonas putida
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
|
|