Metabolic engineering of Pseudomonas putida for production of vanillylamine from lignin‐derived substrates

Summary 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 recombi...

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Autores principales:	João Heitor Colombelli Manfrão‐Netto, Fredrik Lund, Nina Muratovska, Elin M. Larsson, Nádia Skorupa Parachin, Magnus Carlquist
Formato:	article
Lenguaje:	EN
Publicado:	Wiley 2021
Materias:	Biotechnology TP248.13-248.65
Acceso en línea:	https://doaj.org/article/07d429cc11f6411180fa9ac2276cfd5c
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spelling	oai:doaj.org-article:07d429cc11f6411180fa9ac2276cfd5c2021-11-18T15:39:52ZMetabolic engineering of Pseudomonas putida for production of vanillylamine from lignin‐derived substrates1751-791510.1111/1751-7915.13764https://doaj.org/article/07d429cc11f6411180fa9ac2276cfd5c2021-11-01T00:00:00Zhttps://doi.org/10.1111/1751-7915.13764https://doaj.org/toc/1751-7915Summary 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.João Heitor Colombelli Manfrão‐NettoFredrik LundNina MuratovskaElin M. LarssonNádia Skorupa ParachinMagnus CarlquistWileyarticleBiotechnologyTP248.13-248.65ENMicrobial Biotechnology, Vol 14, Iss 6, Pp 2448-2462 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Biotechnology TP248.13-248.65
spellingShingle	Biotechnology TP248.13-248.65 João Heitor Colombelli Manfrão‐Netto Fredrik Lund Nina Muratovska Elin M. Larsson Nádia Skorupa Parachin Magnus Carlquist Metabolic engineering of Pseudomonas putida for production of vanillylamine from lignin‐derived substrates
description	Summary 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.
format	article
author	João Heitor Colombelli Manfrão‐Netto Fredrik Lund Nina Muratovska Elin M. Larsson Nádia Skorupa Parachin Magnus Carlquist
author_facet	João Heitor Colombelli Manfrão‐Netto Fredrik Lund Nina Muratovska Elin M. Larsson Nádia Skorupa Parachin Magnus Carlquist
author_sort	João Heitor Colombelli Manfrão‐Netto
title	Metabolic engineering of Pseudomonas putida for production of vanillylamine from lignin‐derived substrates
title_short	Metabolic engineering of Pseudomonas putida for production of vanillylamine from lignin‐derived substrates
title_full	Metabolic engineering of Pseudomonas putida for production of vanillylamine from lignin‐derived substrates
title_fullStr	Metabolic engineering of Pseudomonas putida for production of vanillylamine from lignin‐derived substrates
title_full_unstemmed	Metabolic engineering of Pseudomonas putida for production of vanillylamine from lignin‐derived substrates
title_sort	metabolic engineering of pseudomonas putida for production of vanillylamine from lignin‐derived substrates
publisher	Wiley
publishDate	2021
url	https://doaj.org/article/07d429cc11f6411180fa9ac2276cfd5c
work_keys_str_mv	AT joaoheitorcolombellimanfraonetto metabolicengineeringofpseudomonasputidaforproductionofvanillylaminefromligninderivedsubstrates AT fredriklund metabolicengineeringofpseudomonasputidaforproductionofvanillylaminefromligninderivedsubstrates AT ninamuratovska metabolicengineeringofpseudomonasputidaforproductionofvanillylaminefromligninderivedsubstrates AT elinmlarsson metabolicengineeringofpseudomonasputidaforproductionofvanillylaminefromligninderivedsubstrates AT nadiaskorupaparachin metabolicengineeringofpseudomonasputidaforproductionofvanillylaminefromligninderivedsubstrates AT magnuscarlquist metabolicengineeringofpseudomonasputidaforproductionofvanillylaminefromligninderivedsubstrates
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Metabolic engineering of Pseudomonas putida for production of vanillylamine from lignin‐derived substrates

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