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Periplasmic expression of Pseudomonas fluorescens peroxidase Dyp1B and site-directed mutant Dyp1B enzymes enhances polymeric lignin degradation activity in Pseudomonas putida KT2440
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Ehibhatiomhan, Austine O., Rahmanpour, Rahman, Farnaud, Sebastien, Bugg, Timothy D. H. and Mendel-Williams, Sharon (2023) Periplasmic expression of Pseudomonas fluorescens peroxidase Dyp1B and site-directed mutant Dyp1B enzymes enhances polymeric lignin degradation activity in Pseudomonas putida KT2440. Enzyme & Microbial Technology, 162 . 110147. doi:10.1016/j.enzmictec.2022.110147 ISSN 0141-0229.
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Official URL: https://doi.org/10.1016/j.enzmictec.2022.110147
Abstract
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.
Item Type: | Journal Article | |||||||||
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Subjects: | Q Science > QD Chemistry Q Science > QK Botany Q Science > QP Physiology Q Science > QR Microbiology T Technology > TN Mining engineering. Metallurgy |
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Chemistry | |||||||||
Library of Congress Subject Headings (LCSH): | Peroxidase, Manganese oxides, Manganese -- Oxidation, Lignin -- Biodegradation, Pseudomonas fluorescens , Pseudomonas | |||||||||
Journal or Publication Title: | Enzyme & Microbial Technology | |||||||||
Publisher: | Elsevier | |||||||||
ISSN: | 0141-0229 | |||||||||
Official Date: | January 2023 | |||||||||
Dates: |
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Volume: | 162 | |||||||||
Number of Pages: | 8 | |||||||||
Article Number: | 110147 | |||||||||
DOI: | 10.1016/j.enzmictec.2022.110147 | |||||||||
Status: | Peer Reviewed | |||||||||
Publication Status: | Published | |||||||||
Access rights to Published version: | Open Access (Creative Commons) | |||||||||
Date of first compliant deposit: | 25 October 2022 | |||||||||
Date of first compliant Open Access: | 23 November 2022 | |||||||||
RIOXX Funder/Project Grant: |
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