Metabolic engineering of beta-oxidation in Penicillium chrysogenum for improved semi-synthetic cephalosporin biosynthesis

Tânia Veiga; Andreas K Gombert; Nils Landes; Maarten D Verhoeven; Jan A K W Kiel; Arjen M Krikken; Jeroen G Nijland; Hesselien Touw; Marijke A H Luttik; John C van der Toorn; Arnold J M Driessen; Roel A L Bovenberg; Marco A van den Berg; Ida J van der Klei; Jack T Pronk; Jean-Marc Daran

doi:10.1016/j.ymben.2012.02.004

Metabolic engineering of beta-oxidation in Penicillium chrysogenum for improved semi-synthetic cephalosporin biosynthesis

Tânia Veiga, Andreas K Gombert, Nils Landes, Maarten D Verhoeven, Jan A K W Kiel, Arjen M Krikken, Jeroen G Nijland, Hesselien Touw, Marijke A H Luttik, John C van der Toorn, Arnold J M Driessen, Roel A L Bovenberg, Marco A van den Berg, Ida J van der Klei, Jack T Pronk, Jean-Marc Daran^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

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Abstract

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.

Original language	English
Pages (from-to)	437-448
Number of pages	12
Journal	Metabolic Engineering
Volume	14
Issue number	4
DOIs	https://doi.org/10.1016/j.ymben.2012.02.004
Publication status	Published - Jul-2012

Keywords

Penicillium chrysogenum
beta-lactams
Cephalosporins
beta-oxidation
Adipic acid
Metabolic engineering
CHAIN DICARBOXYLIC-ACIDS
CLAVULIGERUS CEFE GENE
ACYL-COA OXIDASE
C-14 ADIPIC ACID
SACCHAROMYCES-CEREVISIAE
ASPERGILLUS-NIDULANS
STREPTOMYCES-CLAVULIGERUS
FATTY-ACIDS
HUMAN-URINE
PEROXISOME PROLIFERATION

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Veiga, T., Gombert, A. K., Landes, N., Verhoeven, M. D., Kiel, J. A. K. W., Krikken, A. M., Nijland, J. G., Touw, H., Luttik, M. A. H., van der Toorn, J. C., Driessen, A. J. M., Bovenberg, R. A. L., van den Berg, M. A., van der Klei, I. J., Pronk, J. T., & Daran, J.-M. (2012). Metabolic engineering of beta-oxidation in Penicillium chrysogenum for improved semi-synthetic cephalosporin biosynthesis. Metabolic Engineering, 14(4), 437-448. https://doi.org/10.1016/j.ymben.2012.02.004

@article{ae09b0c3e87347fea3b1e37fcdcd9ae5,

title = "Metabolic engineering of beta-oxidation in Penicillium chrysogenum for improved semi-synthetic cephalosporin biosynthesis",

abstract = "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.",

keywords = "Penicillium chrysogenum, beta-lactams, Cephalosporins, beta-oxidation, Adipic acid, Metabolic engineering, CHAIN DICARBOXYLIC-ACIDS, CLAVULIGERUS CEFE GENE, ACYL-COA OXIDASE, C-14 ADIPIC ACID, SACCHAROMYCES-CEREVISIAE, ASPERGILLUS-NIDULANS, STREPTOMYCES-CLAVULIGERUS, FATTY-ACIDS, HUMAN-URINE, PEROXISOME PROLIFERATION",

author = "T{\^a}nia Veiga and Gombert, {Andreas K} and Nils Landes and Verhoeven, {Maarten D} and Kiel, {Jan A K W} and Krikken, {Arjen M} and Nijland, {Jeroen G} and Hesselien Touw and Luttik, {Marijke A H} and {van der Toorn}, {John C} and Driessen, {Arnold J M} and Bovenberg, {Roel A L} and {van den Berg}, {Marco A} and {van der Klei}, {Ida J} and Pronk, {Jack T} and Jean-Marc Daran",

year = "2012",

month = jul,

doi = "10.1016/j.ymben.2012.02.004",

language = "English",

volume = "14",

pages = "437--448",

journal = "Metabolic Engineering",

issn = "1096-7176",

publisher = "ACADEMIC PRESS INC ELSEVIER SCIENCE",

number = "4",

}

Veiga, T, Gombert, AK, Landes, N, Verhoeven, MD, Kiel, JAKW , Krikken, AM , Nijland, JG, Touw, H, Luttik, MAH, van der Toorn, JC, Driessen, AJM , Bovenberg, RAL, van den Berg, MA, van der Klei, IJ, Pronk, JT & Daran, J-M 2012, 'Metabolic engineering of beta-oxidation in Penicillium chrysogenum for improved semi-synthetic cephalosporin biosynthesis', Metabolic Engineering, vol. 14, no. 4, pp. 437-448. https://doi.org/10.1016/j.ymben.2012.02.004

TY - JOUR

T1 - Metabolic engineering of beta-oxidation in Penicillium chrysogenum for improved semi-synthetic cephalosporin biosynthesis

AU - Veiga, Tânia

AU - Gombert, Andreas K

AU - Landes, Nils

AU - Verhoeven, Maarten D

AU - Kiel, Jan A K W

AU - Krikken, Arjen M

AU - Nijland, Jeroen G

AU - Touw, Hesselien

AU - Luttik, Marijke A H

AU - van der Toorn, John C

AU - Driessen, Arnold J M

AU - Bovenberg, Roel A L

AU - van den Berg, Marco A

AU - van der Klei, Ida J

AU - Pronk, Jack T

AU - Daran, Jean-Marc

PY - 2012/7

Y1 - 2012/7

N2 - 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.

AB - 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.

KW - Penicillium chrysogenum

KW - beta-lactams

KW - Cephalosporins

KW - beta-oxidation

KW - Adipic acid

KW - Metabolic engineering

KW - CHAIN DICARBOXYLIC-ACIDS

KW - CLAVULIGERUS CEFE GENE

KW - ACYL-COA OXIDASE

KW - C-14 ADIPIC ACID

KW - SACCHAROMYCES-CEREVISIAE

KW - ASPERGILLUS-NIDULANS

KW - STREPTOMYCES-CLAVULIGERUS

KW - FATTY-ACIDS

KW - HUMAN-URINE

KW - PEROXISOME PROLIFERATION

U2 - 10.1016/j.ymben.2012.02.004

DO - 10.1016/j.ymben.2012.02.004

M3 - Article

C2 - 22525490

SN - 1096-7176

VL - 14

SP - 437

EP - 448

JO - Metabolic Engineering

JF - Metabolic Engineering

IS - 4

ER -

Metabolic engineering of beta-oxidation in Penicillium chrysogenum for improved semi-synthetic cephalosporin biosynthesis

Abstract

Keywords

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