The Penicillium chrysogenum aclA gene encodes a broad-substrate-specificity acyl-coenzyme A ligase involved in activation of adipic acid, a side-chain precursor for cephem antibiotics

Koetsier, Martijn J.; Gombert, Andreas; Fekken, Susan; Bovenberg, Roel A. L.; Berg, M. A.; Kiel, Jan A.K.W.; Jekel, Peter A.; Janssen, Dick B.; Pronk, Jack T.; Klei, Ida J. van der; Daran, Jean‐Marc

doi:10.1016/j.fgb.2009.10.003

Cited by 31 publications

(30 citation statements)

References 53 publications

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“…For each strain, transcriptome analysis was performed from at least two independent replicate chemostat cultures. The average coefficient of variation for transcriptome data from replicated cultures was below 20%, in accordance with previous chemostat-based transcriptome analyses in P. chrysogenum (Douma et al, 2011;Gombert et al, 2011;Harris et al, 2009aHarris et al, , 2009bKoetsier et al, 2010;Snoek et al, 2009;van den Berg et al, 2008).…”

Section: Chemostat-based Transcriptome Analysis: Data Quality and Ovesupporting

confidence: 90%

Impact of Velvet Complex on Transcriptome and Penicillin G Production in Glucose-Limited Chemostat Cultures of a β-Lactam High-ProducingPenicillium chrysogenumStrain

Veiga

Nijland

Driessen

et al. 2012

OMICS: A Journal of Integrative Biology

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The multicomponent global regulator Velvet complex has been identified as a key regulator of secondary metabolite production in Aspergillus and Penicillium species. Previous work indicated a massive impact of PcvelA and PclaeA deletions on penicillin production in prolonged batch cultures of P. chrysogenum, as well as substantial changes in transcriptome. The present study investigated the impact of these mutations on product formation and genome-wide transcript profiles under glucose-limited aerobic conditions, relevant for industrial production of b-lactams. Predicted amino acid sequences of PcVelA and PcLaeA in this strain were identical to those in its ancestor Wisconsin54-1255. Controls were performed to rule out transformation-associated loss of penicillin-biosynthesis clusters. The correct PcvelA and PclaeA deletion strains revealed a small reduction of penicillin G productivity relative to the reference strain, which is a much smaller reduction than previously reported for prolonged batch cultures of similar P. chrysogenum mutants. Chemostat-based transcriptome analysis yielded only 23 genes with a consistent differential response in the PcvelAD and PclaeAD mutants when grown in the absence of the penicillin G side-chain precursor phenylacetic acid. Eleven of these genes belonged to two small gene clusters, one of which contained a gene with high homology to the aristolochene synthase. These results provide a clear caveat that the impact of the Velvet complex on secondary metabolism in filamentous fungi is strongly context dependent.

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Section: Chemostat-based Transcriptome Analysis: Data Quality and Ovesupporting

confidence: 90%

Impact of Velvet Complex on Transcriptome and Penicillin G Production in Glucose-Limited Chemostat Cultures of a β-Lactam High-ProducingPenicillium chrysogenumStrain

Veiga

Nijland

Driessen

et al. 2012

OMICS: A Journal of Integrative Biology

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“…The average coefficient of variation for biological replicates (triplicates for the carbon-limited cultures with ammonium sul- a Three different network scenario hypotheses were employed to obtain an optimal fit for flux distribution: scenario i, phenylalanine was incorporated into protein, incorporated into penicillin G, metabolized via the homogentisate pathway, or metabolized via tyrosine; scenario ii, an unidentified pathway for phenylalanine metabolism, which did not involve any of the measured metabolites, was introduced into the model together with exchanges of phenylalanine and tyrosine with the protein pool (synthesis and degradation); and scenario iii, hypothetical hydroxylase activities that convert phenylpyruvate to 4-hydroxyphenylpyruvate and phenylacetaldehyde to 4-hydroxyphenylacetaldehyde were included. (14,26,27,36,68,71).…”

Section: Resultsmentioning

confidence: 99%

Resolving Phenylalanine Metabolism Sheds Light on Natural Synthesis of Penicillin G in Penicillium chrysogenum

et al. 2012

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The industrial production of penicillin G by Penicillium chrysogenum requires the supplementation of the growth medium with the side chain precursor phenylacetate. The growth of P. chrysogenum with phenylalanine as the sole nitrogen source resulted in the extracellular production of phenylacetate and penicillin G. To analyze this natural pathway for penicillin G production, chemostat cultures were switched to [U-13 C]phenylalanine as the nitrogen source. The quantification and modeling of the dynamics of labeled metabolites indicated that phenylalanine was (i) incorporated in nascent protein, (ii) transaminated to phenylpyruvate and further converted by oxidation or by decarboxylation, and (iii) hydroxylated to tyrosine and subsequently metabolized via the homogentisate pathway. The involvement of the homogentisate pathway was supported by the comparative transcriptome analysis of P. chrysogenum cultures grown with phenylalanine and with (NH 4 ) 2 SO 4 as the nitrogen source. This transcriptome analysis also enabled the identification of two putative 2-oxo acid decarboxylase genes (Pc13g9300 and Pc18g01490). cDNAs of both genes were cloned and expressed in the 2-oxo-acid-decarboxylase-free Saccharomyces cerevisiae strain CEN.PK711-7C (pdc1 pdc5 pdc6⌬ aro10⌬ thi3⌬). The introduction of Pc13g09300 restored the growth of this S. cerevisiae mutant on glucose and phenylalanine, thereby demonstrating that Pc13g09300 encodes a dual-substrate pyruvate and phenylpyruvate decarboxylase, which plays a key role in an Ehrlich-type pathway for the production of phenylacetate in P. chrysogenum. These results provide a basis for the metabolic engineering of P. chrysogenum for the production of the penicillin G side chain precursor phenylacetate.

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“…The use of transcriptome analyses on glucose-limited chemostat cultures of these strains, grown in the presence and absence of PAA, showed that the homogentisate pathway for PAA catabolism was strongly transcriptionally up-regulated in PAA-fed cultures; and that several genes involved in nitrogen and sulfur metabolism were transcriptionally up-regulated only under penicillin G producing conditions suggesting a drain of amino acid precursor pools. Most recently this group has again used chemostatbased transcriptome analysis to identify the gene encoding the acyl-CoA ligase involved in the activation of adipate prior to its incorporation into the b-lactam backbone [105]. The results from both of these studies indicate clear-cut targets for metabolic engineering, in the former case genes beyond those of the b-lactam pathway per se, and in the latter case for exploring the production of cephalosporin analogs.…”

Section: Industrial Applicationsmentioning

confidence: 99%

The renaissance of continuous culture in the post-genomics age

Bull

2010

J Ind Microbiol Biotechnol

108

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The development of continuous culture techniques 60 years ago and the subsequent formulation of theory and the diversification of experimental systems revolutionised microbiology and heralded a unique period of innovative research. Then, progressively, molecular biology and thence genomics and related high-information-density omics technologies took centre stage and microbial growth physiology in general faded from educational programmes and research funding priorities alike. However, there has been a gathering appreciation over the past decade that if the claims of systems biology are going to be realised, they will have to be based on rigorously controlled and reproducible microbial and cell growth platforms. This revival of continuous culture will be long lasting because its recognition as the growth system of choice is firmly established. The purpose of this review, therefore, is to remind microbiologists, particularly those new to continuous culture approaches, of the legacy of what I call the first age of continuous culture, and to explore a selection of researches that are using these techniques in this post-genomics age. The review looks at the impact of continuous culture across a comprehensive range of microbiological research and development. The ability to establish (quasi-) steady state conditions is a frequently stated advantage of continuous cultures thereby allowing environmental parameters to be manipulated without causing concomitant changes in the specific growth rate. However, the use of continuous cultures also enables the critical study of specified transition states and chemical, physical or biological perturbations. Such dynamic analyses enhance our understanding of microbial ecology and microbial pathology for example, and offer a wider scope for innovative drug discovery; they also can inform the optimization of batch and fed-batch operations that are characterized by sequential transitions states.

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The Penicillium chrysogenum aclA gene encodes a broad-substrate-specificity acyl-coenzyme A ligase involved in activation of adipic acid, a side-chain precursor for cephem antibiotics

Cited by 31 publications

References 53 publications

Impact of Velvet Complex on Transcriptome and Penicillin G Production in Glucose-Limited Chemostat Cultures of a β-Lactam High-ProducingPenicillium chrysogenumStrain

Impact of Velvet Complex on Transcriptome and Penicillin G Production in Glucose-Limited Chemostat Cultures of a β-Lactam High-ProducingPenicillium chrysogenumStrain

Resolving Phenylalanine Metabolism Sheds Light on Natural Synthesis of Penicillin G in Penicillium chrysogenum

The renaissance of continuous culture in the post-genomics age

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