Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
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Tryptophan-Based Hyperproduction of Bioindigo by Combinatorial Overexpression of Two Different Tryptophan Transporters

  • Hyun Jin Kim (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Sion Ham (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Nara-Shin (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Jeong Hyeon Hwang (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Suk Jin Oh (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Tae-Rim Choi (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Jeong Chan Joo (Department of Chemical Engineering, Kyung Hee University) ;
  • Shashi Kant Bhatia (Department of Biological Engineering, College of Engineering, Konkuk University) ;
  • Yung-Hun Yang (Department of Biological Engineering, College of Engineering, Konkuk University)
  • Received : 2023.08.21
  • Accepted : 2023.11.22
  • Published : 2024.04.28
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Abstract

Indigo is a valuable, natural blue dye that has been used for centuries in the textile industry. The large-scale commercial production of indigo relies on its extraction from plants and chemical synthesis. Studies are being conducted to develop methods for environment-friendly and sustainable production of indigo using genetically engineered microbes. Here, to enhance the yield of bioindigo from an E. coli whole-cell system containing tryptophanase (TnaA) and flavin-containing monooxygenase (FMO), we evaluated tryptophan transporters to improve the transport of aromatic compounds, such as indole and tryptophan, which are not easily soluble and passable through cell walls. Among the three transporters, Mtr, AroP, and TnaB, AroP enhanced indigo production the most. The combination of each transporter with AroP was also evaluated, and the combination of AroP and TnaB showed the best performance compared to the single transporters and two transporters. Bioindigo production was then optimized by examining the culture medium, temperature, isopropyl β-D-1-thiogalactopyranoside concentration, shaking speed (rpm), and pH. The novel strain containing aroP and tnaB plasmid with tnaA and FMO produced 8.77 mM (2.3 g/l) of bioindigo after 66 h of culture. The produced bioindigo was further recovered using a simple method and used as a watercolor dye, showing good mixing with other colors and color retention for a relatively long time. This study presents an effective strategy for enhancing indigo production using a combination of transporters.

Keywords

Acknowledgement

This research was supported by the National Research Foundation of Korea (NRF), the Ministry of Science and ICT (NRF-2022R1A2C2003138, NRF-2022M3I3A1082545) and the R&D Program of MOTIE/KEIT (Grant No. 20009508 and 20014350). This paper was also supported by the Konkuk University Researcher Fund in 2023.

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