Metabolic engineering of Pseudomonas putida for production of vanillylamine from lignin-derived substrates
(2021) In Microbial Biotechnology 14(6). p.2448-2462- Abstract
- Whole-cell bioconversion of technical lignins using Pseudomonas putida strains
overexpressing amine transaminases (ATAs) has the potential to become
an eco-efficient route to produce phenolic amines. Here, a novel cell
growth-based screening method to evaluate the in vivo activity of recombinant ATAs towards vanillylamine in P. putida KT2440 was developed. It allowed the identification of the native enzyme Pp-SpuC-II and ATA from Chromobacterium violaceum (Cv-ATA) as highly active towards vanillylamine in vivo. Overexpression of Pp-SpuC-II and Cv-ATA
in the strain GN442ΔPP_2426, previously engineered for reduced vanillin
assimilation, resulted in 94- and... (More) - Whole-cell bioconversion of technical lignins using Pseudomonas putida strains
overexpressing amine transaminases (ATAs) has the potential to become
an eco-efficient route to produce phenolic amines. Here, a novel cell
growth-based screening method to evaluate the in vivo activity of recombinant ATAs towards vanillylamine in P. putida KT2440 was developed. It allowed the identification of the native enzyme Pp-SpuC-II and ATA from Chromobacterium violaceum (Cv-ATA) as highly active towards vanillylamine in vivo. Overexpression of Pp-SpuC-II and Cv-ATA
in the strain GN442ΔPP_2426, previously engineered for reduced vanillin
assimilation, resulted in 94- and 92-fold increased specific
transaminase activity, respectively. Whole-cell bioconversion of
vanillin yielded 0.70 ± 0.20 mM and 0.92 ± 0.30 mM vanillylamine, for Pp-SpuC-II and Cv-ATA,
respectively. Still, amine production was limited by a substantial
re-assimilation of the product and formation of the by-products vanillic
acid and vanillyl alcohol. Concomitant overexpression of Cv-ATA and alanine dehydrogenase from Bacillus subtilis
increased the production of vanillylamine with ammonium as the only
nitrogen source and a reduction in the amount of amine product
re-assimilation. Identification and deletion of additional native genes
encoding oxidoreductases acting on vanillin are crucial engineering
targets for further improvement. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/f4493d8f-b033-4c4b-b0fa-f457de126ab4
- author
- Heitor Colombelli Manfrão-Netto, João ; Lund, Fredrik LU ; Muratovska, Nina LU ; Larsson, Elin M. ; Skorupa Parachin, Nádia and Carlquist, Magnus LU
- organization
- publishing date
- 2021-02-03
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Microbial Biotechnology
- volume
- 14
- issue
- 6
- pages
- 15 pages
- publisher
- John Wiley & Sons Inc.
- external identifiers
-
- pmid:33533574
- scopus:85100356086
- ISSN
- 1751-7907
- DOI
- 10.1111/1751-7915.13764
- project
- Exploration of Brazilian biodiversity for new biosynthetic pathways applied to lignin valorization
- language
- English
- LU publication?
- yes
- id
- f4493d8f-b033-4c4b-b0fa-f457de126ab4
- date added to LUP
- 2022-02-07 09:18:05
- date last changed
- 2023-01-24 23:06:13
@article{f4493d8f-b033-4c4b-b0fa-f457de126ab4, abstract = {{Whole-cell bioconversion of technical lignins using <i>Pseudomonas putida strains</i><br> overexpressing amine transaminases (ATAs) has the potential to become <br> an eco-efficient route to produce phenolic amines. Here, a novel cell <br> growth-based screening method to evaluate the <i>in vivo</i> activity of recombinant ATAs towards vanillylamine in <i>P. putida</i> KT2440 was developed. It allowed the identification of the native enzyme Pp-SpuC-II and ATA from <i>Chromobacterium violaceum</i> (<i>Cv-ATA</i>) as highly active towards vanillylamine <i>in vivo</i>. Overexpression of <i>Pp-SpuC-II</i> and <i>Cv-ATA</i><br> in the strain GN442ΔPP_2426, previously engineered for reduced vanillin<br> assimilation, resulted in 94- and 92-fold increased specific <br> transaminase activity, respectively. Whole-cell bioconversion of <br> vanillin yielded 0.70 ± 0.20 mM and 0.92 ± 0.30 mM vanillylamine, for <i>Pp-SpuC-II</i> and <i>Cv-ATA</i>,<br> respectively. Still, amine production was limited by a substantial <br> re-assimilation of the product and formation of the by-products vanillic<br> acid and vanillyl alcohol. Concomitant overexpression of <i>Cv-ATA</i> and alanine dehydrogenase from <i>Bacillus subtilis</i><br> increased the production of vanillylamine with ammonium as the only <br> nitrogen source and a reduction in the amount of amine product <br> re-assimilation. Identification and deletion of additional native genes <br> encoding oxidoreductases acting on vanillin are crucial engineering <br> targets for further improvement.}}, author = {{Heitor Colombelli Manfrão-Netto, João and Lund, Fredrik and Muratovska, Nina and Larsson, Elin M. and Skorupa Parachin, Nádia and Carlquist, Magnus}}, issn = {{1751-7907}}, language = {{eng}}, month = {{02}}, number = {{6}}, pages = {{2448--2462}}, publisher = {{John Wiley & Sons Inc.}}, series = {{Microbial Biotechnology}}, title = {{Metabolic engineering of <i>Pseudomonas putida</i> for production of vanillylamine from lignin-derived substrates}}, url = {{http://dx.doi.org/10.1111/1751-7915.13764}}, doi = {{10.1111/1751-7915.13764}}, volume = {{14}}, year = {{2021}}, }