Metabolic engineering of beta-oxidation in Penicillium chrysogenum for improved semi-synthetic cephalosporin biosynthesis
| Autor: | Maarten D Verhoeven, Jan A.K.W. Kiel, Jean-Marc Daran, Jack T. Pronk, Arjen M Krikken, Roel A. L. Bovenberg, Tania Veiga, Marco A. van den Berg, Hesselien Touw, John C. van der Toorn, Nils Landes, Arnold J. M. Driessen, Marijke A. H. Luttik, Jeroen G. Nijland, Ida J. van der Klei, Andreas Karoly Gombert |
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| Přispěvatelé: | Molecular Cell Biology, Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology |
| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Adipates
Dehydrogenase Bioengineering CHAIN DICARBOXYLIC-ACIDS Penicillium chrysogenum Applied Microbiology and Biotechnology beta-oxidation Metabolic engineering SACCHAROMYCES-CEREVISIAE chemistry.chemical_compound Acyl-CoA Dehydrogenases Adipate Gene Expression Regulation Fungal PEROXISOME PROLIFERATION Peroxisomes CLAVULIGERUS CEFE GENE ACYL-COA OXIDASE ?-lactams Oxidase test Adipic acid biology C-14 ADIPIC ACID beta-lactams Peroxisome HUMAN-URINE biology.organism_classification Mitochondria Cephalosporins Glucose STREPTOMYCES-CLAVULIGERUS chemistry Biochemistry ASPERGILLUS-NIDULANS ?-oxidation FATTY-ACIDS Energy source Transcriptome Oxidation-Reduction Gene Deletion Biotechnology |
| Zdroj: | Metabolic Engineering, 14 (4), 2012 Metabolic Engineering, 14(4), 437-448. ACADEMIC PRESS INC ELSEVIER SCIENCE |
| ISSN: | 1096-7176 |
| Popis: | Industrial production of semi-synthetic cephalosporins by Penicillium chrysogenum requires supplementation of the growth media with the side-chain precursor adipic acid. In glucose-limited chemostat cultures of P. chrysogenum, up to 88% of the consumed adipic acid was not recovered in cephalosporinrelated products, but used as an additional carbon and energy source for growth. This low efficiency of side-chain precursor incorporation provides an economic incentive for studying and engineering the metabolism of adipic acid in P. cluysogenum. Chemostat-based transcriptome analysis in the presence and absence of adipic acid confirmed that adipic acid metabolism in this fungus occurs via beta-oxidation. A set of 52 adipate-responsive genes included six putative genes for acyl-CoA oxidases and dehydrogenases, enzymes responsible for the first step of beta-oxidation. Subcellular localization of the differentially expressed acyl-CoA oxidases and dehydrogenases revealed that the oxidases were exclusively targeted to peroxisomes, while the dehydrogenases were found either in peroxisomes or in mitochondria. Deletion of the genes encoding the peroxisomal acyl-CoA oxidase Pc20g01800 and the mitochondrial acyl-CoA dehydrogenase Pc20g07920 resulted in a 1.6- and 3.7-fold increase in the production of the semi-synthetic cephalosporin intermediate adipoyl-6-APA, respectively. The deletion strains also showed reduced adipate consumption compared to the reference strain, indicating that engineering of the first step of beta-oxidation successfully redirected a larger fraction of adipic acid towards cephalosporin biosynthesis. (C) 2012 Elsevier Inc. All rights reserved. |
| Databáze: | OpenAIRE |
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