Production of Vanillin From Ferulic Acid by Pseudomonas putida KT2440 Using Metabolic Engineering and In Situ Product Recovery

ABSTRACT Vanillin is the most in‐demand flavouring compound in the world and because vanillin extracted from vanilla pods cannot meet the global demand, most vanillin on the market today is chemically synthesised. Increasing demands by consumers for natural ingredients have inspired efforts to devel...

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Main Authors: Ilona A. Ruhl, Sean P. Woodworth, Stefan J. Haugen, Hannah M. Alt, Gregg T. Beckham, Christopher W. Johnson
Format: Article
Language:English
Published: Wiley 2025-05-01
Series:Microbial Biotechnology
Subjects:
Online Access:https://doi.org/10.1111/1751-7915.70152
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author Ilona A. Ruhl
Sean P. Woodworth
Stefan J. Haugen
Hannah M. Alt
Gregg T. Beckham
Christopher W. Johnson
author_facet Ilona A. Ruhl
Sean P. Woodworth
Stefan J. Haugen
Hannah M. Alt
Gregg T. Beckham
Christopher W. Johnson
author_sort Ilona A. Ruhl
collection DOAJ
description ABSTRACT Vanillin is the most in‐demand flavouring compound in the world and because vanillin extracted from vanilla pods cannot meet the global demand, most vanillin on the market today is chemically synthesised. Increasing demands by consumers for natural ingredients have inspired efforts to develop vanillin derived from microbial sources. These efforts have been challenged by low titers, likely caused by the toxicity of vanillin to most microbial biocatalysts. In this study, we engineered a Pseudomonas putida KT2440‐derived strain that accumulated vanillin from ferulic acid to 0.64 g/L. To increase the overall titre, we applied a hydrophobic polystyrene‐based resin to vanillin‐accumulating cultures, which enabled an increase in total vanillin recovery to an apparent titre of 3.35 g/L. This study demonstrates that P. putida can accumulate vanillin from ferulic acid to higher titers when vanillin is removed from the cultivation medium, mitigating its toxicity.
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series Microbial Biotechnology
spelling doaj-art-7ab58a545abf47b896f2c4742bbb3c432025-08-20T01:57:01ZengWileyMicrobial Biotechnology1751-79152025-05-01185n/an/a10.1111/1751-7915.70152Production of Vanillin From Ferulic Acid by Pseudomonas putida KT2440 Using Metabolic Engineering and In Situ Product RecoveryIlona A. Ruhl0Sean P. Woodworth1Stefan J. Haugen2Hannah M. Alt3Gregg T. Beckham4Christopher W. Johnson5Renewable Resources and Enabling Sciences Center National Renewable Energy Laboratory Golden Colorado USARenewable Resources and Enabling Sciences Center National Renewable Energy Laboratory Golden Colorado USARenewable Resources and Enabling Sciences Center National Renewable Energy Laboratory Golden Colorado USARenewable Resources and Enabling Sciences Center National Renewable Energy Laboratory Golden Colorado USARenewable Resources and Enabling Sciences Center National Renewable Energy Laboratory Golden Colorado USARenewable Resources and Enabling Sciences Center National Renewable Energy Laboratory Golden Colorado USAABSTRACT Vanillin is the most in‐demand flavouring compound in the world and because vanillin extracted from vanilla pods cannot meet the global demand, most vanillin on the market today is chemically synthesised. Increasing demands by consumers for natural ingredients have inspired efforts to develop vanillin derived from microbial sources. These efforts have been challenged by low titers, likely caused by the toxicity of vanillin to most microbial biocatalysts. In this study, we engineered a Pseudomonas putida KT2440‐derived strain that accumulated vanillin from ferulic acid to 0.64 g/L. To increase the overall titre, we applied a hydrophobic polystyrene‐based resin to vanillin‐accumulating cultures, which enabled an increase in total vanillin recovery to an apparent titre of 3.35 g/L. This study demonstrates that P. putida can accumulate vanillin from ferulic acid to higher titers when vanillin is removed from the cultivation medium, mitigating its toxicity.https://doi.org/10.1111/1751-7915.70152bioconversionferulic acidin situ product recoverymetabolic engineeringPseudomonas putida KT2440vanillin
spellingShingle Ilona A. Ruhl
Sean P. Woodworth
Stefan J. Haugen
Hannah M. Alt
Gregg T. Beckham
Christopher W. Johnson
Production of Vanillin From Ferulic Acid by Pseudomonas putida KT2440 Using Metabolic Engineering and In Situ Product Recovery
Microbial Biotechnology
bioconversion
ferulic acid
in situ product recovery
metabolic engineering
Pseudomonas putida KT2440
vanillin
title Production of Vanillin From Ferulic Acid by Pseudomonas putida KT2440 Using Metabolic Engineering and In Situ Product Recovery
title_full Production of Vanillin From Ferulic Acid by Pseudomonas putida KT2440 Using Metabolic Engineering and In Situ Product Recovery
title_fullStr Production of Vanillin From Ferulic Acid by Pseudomonas putida KT2440 Using Metabolic Engineering and In Situ Product Recovery
title_full_unstemmed Production of Vanillin From Ferulic Acid by Pseudomonas putida KT2440 Using Metabolic Engineering and In Situ Product Recovery
title_short Production of Vanillin From Ferulic Acid by Pseudomonas putida KT2440 Using Metabolic Engineering and In Situ Product Recovery
title_sort production of vanillin from ferulic acid by pseudomonas putida kt2440 using metabolic engineering and in situ product recovery
topic bioconversion
ferulic acid
in situ product recovery
metabolic engineering
Pseudomonas putida KT2440
vanillin
url https://doi.org/10.1111/1751-7915.70152
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