Title
Bioproduction of p-hydroxybenzoate from renewable feedstock by solvent-tolerant Pseudomonas putida S12
Author
Verhoef, S.
Ruijssenaars, H.J.
de Bont, J.A.M.
Wery, J.
TNO Kwaliteit van Leven
Publication year
2007
Abstract
Pseudomonas putida strain S12palB1 was constructed that produces p-hydroxybenzoate from renewable carbon sources via the central metabolite l-tyrosine. P. putida S12palB1 was based on the platform strain P. putida S12TPL3, which has an optimised carbon flux towards l-tyrosine. Phenylalanine ammonia lyase (Pal) was introduced for the conversion of l-tyrosine into p-coumarate, which is further converted into p-hydroxybenzoate by endogenous enzymes. p-Hydroxybenzoate hydroxylase (PobA) was inactivated to prevent the degradation of p-hydroxybenzoate. These modifications resulted in stable accumulation of p-hydroxybenzoate at a yield of 11% (C-mol C-mol-1) on glucose or on glycerol in shake flask cultures. In a glycerol-limited fed-batch fermentation, a final p-hydroxybenzoate concentration of 12.9 mM (1.8 g l-1) was obtained, at a yield of 8.5% (C-mol C-mol-1). A 2-fold increase of the specific p-hydroxybenzoate production rate (qp) was observed when l-tyrosine was supplied to a steady-state C-limited chemostat culture of P. putida S12palB1. This implied that l-tyrosine availability was the bottleneck for p-hydroxybenzoate production under these conditions. When p-coumarate was added instead, qp increased by a factor 4.7, indicating that Pal activity is the limiting factor when sufficient l-tyrosine is available. Thus, two major leads for further improvement of the p-hydroxybenzoate production by P. putida S12palB1 were identified. © 2007 Elsevier B.V. All rights reserved.
Subject
Biology
Biotechnology
Metabolic engineering
p-Hydroxybenzoate
Pseudomonas putida
Renewable feedstock
Solvent-tolerance
Cell culture
Fermentation
Glucose
Glycerol
Metabolites
Metabolic engineering
p-Hydroxybenzoate
Pseudomonas putida
Renewable feedstock
Solvent-tolerance
Aromatic compounds
4 hydroxybenzoate 3 monooxygenase
4 hydroxybenzoic acid
carbon
glucose
glycerol
para coumaric acid
phenylalanine ammonia lyase
solvent
tyrosine
article
bacterial strain
bacterium culture
chemostat
culture technique
enzyme activity
enzyme inactivation
fed batch fermentation
metabolic engineering
metabolite
nonhuman
nutrient
nutrient solution
priority journal
Pseudomonas putida
Pseudomonas putida s12
reaction analysis
shake flask culture
steady state
Pseudomonas putida
To reference this document use:
http://resolver.tudelft.nl/uuid:730aabf4-bca8-4bad-8ebd-c23719d7f723
DOI
https://doi.org/10.1016/j.jbiotec.2007.08.031
TNO identifier
240246
ISSN
0168-1656
Source
Journal of Biotechnology, 132 (1), 49-56
Document type
article