Journal of Microbiology and Biotechnology

  • 제33권4호
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  • Pages.552-558
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  • 2023
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  • 1017-7825(pISSN)
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  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
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A Genetically Encoded Biosensor for the Detection of Levulinic Acid

  • Tae Hyun Kim (Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Seung-Gyun Woo (Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Seong Keun Kim (Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Byeong Hyeon Yoo (Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Jonghyeok Shin (Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Eugene Rha (Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Soo Jung Kim (Department of Integrative Food, Bioscience and Biotechnology, Chonnam National University) ;
  • Kil Koang Kwon (Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Hyewon Lee (Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Haseong Kim (Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Hee-Taek Kim (Department of Food Science and Technology, Chungnam National University) ;
  • Bong-Hyun Sung (Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Seung-Goo Lee (Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB)) ;
  • Dae-Hee Lee (Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB))
  • 투고 : 2023.01.13
  • 심사 : 2023.01.20
  • 발행 : 2023.04.28
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초록

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.

키워드

과제정보

This research was funded by the Bio & Medical Technology Development Program [grant numbers 2020M3A9I5037641 and 2021M3A9I4022731] of the National Research Foundation, funded by the Ministry of Science and ICT of the Republic of Korea and the KRIBB Research Initiative Program [grant number KGM5402322].

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