Development of novel recombinant peroxidase secretion system from Pseudomonas putida for lignin valorisation

  • Bioresour Technol. 2023 Nov:388:129779. doi: 10.1016/j.biortech.2023.129779.
Siseon Lee  1 Minsik Kang  2 Chan-Duck Jung  3 Jung-Hoon Bae  1 Ju Young Lee  3 Young-Kwon Park  4 Jeong Chan Joo  5 Hoyong Kim  3 Jung-Hoon Sohn  2 Bong Hyun Sung  6
Affiliations
  • 1. Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 34141, Republic of Korea.
  • 2. Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 34141, Republic of Korea; Biosystems and Bioengineering Program, Korea University of Science and Technology (UST), Daejeon 34113, Republic of Korea.
  • 3. Center for Bio-based Chemistry, Korea Research Institute of Chemical Technology (KRICT), Ulsan 44429, Republic of Korea.
  • 4. School of Environmental Engineering, University of Seoul, Seoul 02504, Republic of Korea.
  • 5. Department of Biotechnology, The Catholic University of Korea, Bucheon-si, Gyeonggi-do 14662, Republic of Korea.
  • 6. Synthetic Biology Research Center, Korea Research Institute of Bioscience and Biotechnology (KRIBB), Daejeon 34141, Republic of Korea; Biosystems and Bioengineering Program, Korea University of Science and Technology (UST), Daejeon 34113, Republic of Korea; Graduate School of Engineering Biology, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Republic of Korea. Electronic address: [email protected].
Abstract

Pseudomonas putida is a promising strain for lignin valorisation. However, there is a dearth of stable and efficient systems for secreting Enzymes to enhance the process. Therefore, a novel secretion system for Recombinant lignin-depolymerising peroxidase was developed. By adopting a flagellar type III secretion system, P. putida KT-M2, a secretory host strain, was constructed and an optimal secretion signal fusion partner was identified. Application of the dye-decolourising peroxidase of P. putida to this system resulted in efficient oxidation activity of the cell-free supernatant against various chemicals, including lignin model compounds. This peroxidase-secreting strain was examined to confirm its lignin utilisation capability, resulting in the efficient assimilation of various lignin substrates with 2.6-fold higher growth than that of the wild-type strain after 72 h of cultivation. Finally, this novel system will lead efficient Bacterial lignin breakdown and utilization through enzyme secretion, paving the way for sustainable lignin-consolidated bioprocessing.

Keywords
DyP-type peroxidase; Enzyme secretion; Flagellar Type III secretion; Lignin valorization; Pseudomonas putida.
Products