Resolving phenylalanine metabolism sheds light on natural synthesis of penicillin G in Penicillium chrysogenum

Veiga T, Solis-Escalante D, Romagnoli G, Ten Pierick A, Hanemaaijer M, Deshmuhk A, Pronk JT, Daran JM, Wahl SA (2012)

Publication Type: Journal article

Publication year: 2012

Journal

Eukaryotic Cell American Society for Microbiology

Book Volume: 11

Pages Range: 238-249

Journal Issue: 2

DOI: 10.1128/EC.05285-11

Abstract

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- 13C]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 phe-nylpyruvate 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) 2SO 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\ aro10A. 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 phe-nylpyruvate 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. © 2012, American Society for Microbiology. All Rights Reserved.

Authors with CRIS profile

Sebastian Aljoscha Wahl

Involved external institutions

Delft University of Technology (TU Delft) NL Netherlands (NL)

How to cite

APA:

Veiga, T., Solis-Escalante, D., Romagnoli, G., Ten Pierick, A., Hanemaaijer, M., Deshmuhk, A.,... Wahl, S.A. (2012). Resolving phenylalanine metabolism sheds light on natural synthesis of penicillin G in Penicillium chrysogenum. Eukaryotic Cell, 11(2), 238-249. https://dx.doi.org/10.1128/EC.05285-11

MLA:

Veiga, Tania, et al. "Resolving phenylalanine metabolism sheds light on natural synthesis of penicillin G in Penicillium chrysogenum." Eukaryotic Cell 11.2 (2012): 238-249.

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