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Applied Microbiology and Biotechnology

, Volume 99, Issue 22, pp 9577–9589 | Cite as

Increased riboflavin production by manipulation of inosine 5′-monophosphate dehydrogenase in Ashbya gossypii

  • Rubén M. Buey
  • Rodrigo Ledesma-Amaro
  • Mónica Balsera
  • José María de Pereda
  • José Luis Revuelta
Biotechnologically relevant enzymes and proteins
First Online:

Abstract

Guanine nucleotides are the precursors of essential biomolecules including nucleic acids and vitamins such as riboflavin. The enzyme inosine-5′-monophosphate dehydrogenase (IMPDH) catalyzes the ratelimiting step in the guanine nucleotide de novo biosynthetic pathway and plays a key role in controlling the cellular nucleotide pools. Thus, IMPDH is an important metabolic bottleneck in the guanine nucleotide synthesis, susceptible of manipulation by means of metabolic engineering approaches. Herein, we report the functional and structural characterization of the IMPDH enzyme from the industrial fungus Ashbya gossypii. Our data show that the overexpression of the IMPDH gene increases the metabolic flux through the guanine pathway and ultimately enhances 40 % riboflavin production with respect to the wild type. Also, IMPDH disruption results in a 100-fold increase of inosine excretion to the culture media. Our results contribute to the developing metabolic engineering toolbox aiming at improving the production of metabolites with biotechnological interest in A. gossypii.

Keywords

Ashbya gossypii Metabolic engineering Riboflavin Inosine 5′-monophosphate dehydrogenase 

Abstract

Guanine nucleotides are the precursors of essential biomolecules including nucleic acids and vitamins such as riboflavin. The enzyme inosine-5′-monophosphate dehydrogenase (IMPDH) catalyzes the rate-limiting step in the guanine nucleotide de novo biosynthetic pathway and plays a key role in controlling the cellular nucleotide pools. Thus, IMPDH is an important metabolic bottleneck in the guanine nucleotide synthesis, susceptible of manipulation by means of metabolic engineering approaches. Herein, we report the functional and structural characterization of the IMPDH enzyme from the industrial fungus Ashbya gossypii. Our data show that the overexpression of the IMPDH gene increases the metabolic flux through the guanine pathway and ultimately enhances 40 % riboflavin production with respect to the wild type. Also, IMPDH disruption results in a 100-fold increase of inosine excretion to the culture media. Our results contribute to the developing metabolic engineering toolbox aiming at improving the production of metabolites with biotechnological interest in A. gossypii.

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Notes

Acknowledgments

This work was supported by grant BIO2014-56930-P from the Spanish Ministerio de Economía y Competitividad. Rubén M Buey is supported by a “Ramón y Cajal” contract from the Spanish Ministerio de Economía y Competitividad and by a Marie Curie Career Integration Grant (EB-SxIP; FP7-PEOPLE-2011-CIG-293831). We thank MD. Sánchez and S. Domínguez for excellent technical help and N. Skinner for correcting the manuscript. X-ray crystallography data were collected at BL13 (XALOC) beamline at ALBA Synchrotron Light Facility with the collaboration of ALBA staff.

Conflict of interest

RMB, RLA, and JLR declare conflict of interest due to issued and outstanding patent applications covering aspects of this work.

Supplementary material

253_2015_6710_MOESM1_ESM.pdf (3.4 mb)
ESM 1 (PDF 3495 kb)

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© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Rubén M. Buey
    • 1
  • Rodrigo Ledesma-Amaro
  • Mónica Balsera
    • 2
  • José María de Pereda
    • 3
  • José Luis Revuelta
    • 1
  1. 1.Metabolic Engineering Group, Departamento de Microbiología y GenéticaUniversidad de SalamancaSalamancaSpain
  2. 2.Department Abiotic Stress, Instituto de Recursos Naturales y AgrobiologíaConsejo Superior de Investigaciones CientíficasSalamancaSpain
  3. 3.Instituto de Biología Celular y Molecular del Cáncer, Consejo Superior de Investigaciones CientíficasUniversidad de SalamancaSalamancaSpain

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