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Identification of the two-component guaiacol demethylase system from Rhodococcus rhodochrous and expression in Pseudomonas putida EM42 for guaiacol assimilation

García-Hidalgo, Javier LU ; Ravi, Krithika LU ; Kuré, Lise-Lotte; Lidén, Gunnar LU and Gorwa-Grauslund, Marie LU (2019) In AMB Express 9(1).
Abstract

A diversity of softwood lignin depolymerization processes yield guaiacol as the main low molecular weight product. This key aromatic compound can be utilized as a carbon source by several microbial species, most of which are Gram positive bacteria. Microbial degradation of guaiacol is known to proceed initially via demethylation to catechol, and this reaction is catalyzed by cytochrome P450 monooxygenases. These enzymes typically require a set of redox partner proteins, whose number and identities were not described until very recently in the case of guaiacol. In this work we identified two proteins involved in guaiacol demethylation by the actinomycete Rhodococcus rhodochrous. Additionally, we constructed four different polycistronic... (More)

A diversity of softwood lignin depolymerization processes yield guaiacol as the main low molecular weight product. This key aromatic compound can be utilized as a carbon source by several microbial species, most of which are Gram positive bacteria. Microbial degradation of guaiacol is known to proceed initially via demethylation to catechol, and this reaction is catalyzed by cytochrome P450 monooxygenases. These enzymes typically require a set of redox partner proteins, whose number and identities were not described until very recently in the case of guaiacol. In this work we identified two proteins involved in guaiacol demethylation by the actinomycete Rhodococcus rhodochrous. Additionally, we constructed four different polycistronic operons carrying combinations of putative redox partners of this guaiacol demethylation system in an inducible expression plasmid that was introduced into the Gram negative host Pseudomonas putida EM42, and the guaiacol consumption dynamics of each resulting strain were analyzed. All the polycistronic operons, expressing a cytochrome P450 together with a putative ferredoxin reductase from R. rhodochrous and putative ferredoxins from R. rhodochrous or Amycolatopsis ATCC 39116 enabled P. putida EM42 to metabolize and grow on guaiacol as the sole carbon source.

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organization
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Contribution to journal
publication status
published
subject
keywords
Aromatic compound catabolism, Cytochrome P450, Guaiacol demethylase, Lignin, Pseudomonas putida, Rhodococcus rhodochrous
in
AMB Express
volume
9
issue
1
publisher
Springer Open
external identifiers
  • scopus:85062791053
ISSN
2191-0855
DOI
10.1186/s13568-019-0759-8
language
English
LU publication?
yes
id
4d12899e-5bd3-4589-8b33-a7958a5f4db7
date added to LUP
2019-03-19 08:24:41
date last changed
2019-05-21 04:19:45
@article{4d12899e-5bd3-4589-8b33-a7958a5f4db7,
  abstract     = {<p>A diversity of softwood lignin depolymerization processes yield guaiacol as the main low molecular weight product. This key aromatic compound can be utilized as a carbon source by several microbial species, most of which are Gram positive bacteria. Microbial degradation of guaiacol is known to proceed initially via demethylation to catechol, and this reaction is catalyzed by cytochrome P450 monooxygenases. These enzymes typically require a set of redox partner proteins, whose number and identities were not described until very recently in the case of guaiacol. In this work we identified two proteins involved in guaiacol demethylation by the actinomycete Rhodococcus rhodochrous. Additionally, we constructed four different polycistronic operons carrying combinations of putative redox partners of this guaiacol demethylation system in an inducible expression plasmid that was introduced into the Gram negative host Pseudomonas putida EM42, and the guaiacol consumption dynamics of each resulting strain were analyzed. All the polycistronic operons, expressing a cytochrome P450 together with a putative ferredoxin reductase from R. rhodochrous and putative ferredoxins from R. rhodochrous or Amycolatopsis ATCC 39116 enabled P. putida EM42 to metabolize and grow on guaiacol as the sole carbon source.</p>},
  articleno    = {34},
  author       = {García-Hidalgo, Javier and Ravi, Krithika and Kuré, Lise-Lotte and Lidén, Gunnar and Gorwa-Grauslund, Marie},
  issn         = {2191-0855},
  keyword      = {Aromatic compound catabolism,Cytochrome P450,Guaiacol demethylase,Lignin,Pseudomonas putida,Rhodococcus rhodochrous},
  language     = {eng},
  number       = {1},
  publisher    = {Springer Open},
  series       = {AMB Express},
  title        = {Identification of the two-component guaiacol demethylase system from Rhodococcus rhodochrous and expression in Pseudomonas putida EM42 for guaiacol assimilation},
  url          = {http://dx.doi.org/10.1186/s13568-019-0759-8},
  volume       = {9},
  year         = {2019},
}