Expression of lignin-oxidising Pseudomonas fluorescens Dyp1B in the periplasm of Pseudomonas putida KT2440, using a tat fusion construct, was found to lead to enhanced whole cell activity for oxidation of DCP and polymeric lignin substrates. Four amino acid residues predicted to lie at the manganese ion binding site of Pseudomonas fluorescens peroxidase Dyp1B were investigated using site-directed mutagenesis. Mutants H127R and S223A showed 2-fold and 4-fold higher kcat for Mn(II) oxidation respectively, and mutant S223A showed 2-fold enhanced production of low molecular weight phenolic products from a polymeric soda lignin. The mutant Pfl Dyp1B genes were expressed as tat fusions to investigate their effect on lignin oxidation by P. putida KT2440.
|Number of pages||8|
|Journal||Enzyme and Microbial Technology|
|Early online date||20 Oct 2022|
|Publication status||Published - Jan 2023|
Bibliographical noteThis is an open access article under the CC-BY license,
This work was supported by BBSRC ERA-IB research grant BB/M025772/1 , and a Ph.D. studentship from Coventry University (to A.O.E.). We would like to thank Dr. Eduardo Díaz (CSIC Madrid) for the gift of plasmid pIZ1016, Dr. James Williamson (University of Warwick) for practical advice and assistance, and Dr Goran Rashid (University of Warwick) for assistance with HPLC.
This work was supported by BBSRC ERA-IB research grant
- Dye-decolorizing peroxidases
- Lignin degradation
- Manganese oxidation
- Pseudomonas fluorescens
- Pseudomonas putida
ASJC Scopus subject areas
- Applied Microbiology and Biotechnology