Bioproduction of p-hydroxybenzoate from renewable feedstock by solvent-tolerant Pseudomonas putida S12
| Autor: | Harald J. Ruijssenaars, Jan Wery, Suzanne Verhoef, Jan A.M. de Bont |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
DNA
Bacterial Parabens Bioengineering Chemostat Phenylalanine ammonia-lyase Biology Applied Microbiology and Biotechnology Metabolic engineering chemistry.chemical_compound Bioreactors Bioreactor Glycerol Food science DNA Primers Phenylalanine Ammonia-Lyase Base Sequence Pseudomonas putida General Medicine biology.organism_classification Bioproduction Kinetics Biochemistry chemistry Fermentation Solvents Tyrosine Biotechnology Plasmids |
| Zdroj: | Journal of biotechnology. 132(1) |
| ISSN: | 0168-1656 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect