Bioproduction of p-hydroxybenzoate from renewable feedstock by solvent-tolerant Pseudomonas putida S12
article
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.
Topics
BiotechnologyMetabolic engineeringp-HydroxybenzoatePseudomonas putidaRenewable feedstockSolvent-toleranceCell cultureFermentationGlucoseGlycerolMetabolitesMetabolic engineeringp-HydroxybenzoatePseudomonas putidaRenewable feedstockSolvent-toleranceAromatic compounds4 hydroxybenzoate 3 monooxygenase4 hydroxybenzoic acidcarbonglucoseglycerolpara coumaric acidphenylalanine ammonia lyasesolventtyrosinearticlebacterial strainbacterium culturechemostatculture techniqueenzyme activityenzyme inactivationfed batch fermentationmetabolic engineeringmetabolitenonhumannutrientnutrient solutionpriority journalPseudomonas putidaPseudomonas putida s12reaction analysisshake flask culturesteady statePseudomonas putida
TNO Identifier
240246
ISSN
01681656
Source
Journal of Biotechnology, 132(1), pp. 49-56.
Pages
49-56
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