Archives of Microbiology

, Volume 196, Issue 1, pp 25–33 | Cite as

Extracellular expression of glucose inhibition-resistant Cellulomonas flavigena PN-120 β-glucosidase by a diploid strain of Saccharomyces cerevisiae

  • David J. Mendoza-Aguayo
  • Héctor M. Poggi-Varaldo
  • Jaime García-Mena
  • Ana C. Ramos-Valdivia
  • Luis M. Salgado
  • Mayra de la Torre-Martínez
  • Teresa Ponce-NoyolaEmail author
Original Paper


The catalytic fraction of the Cellulomonas flavigena PN-120 oligomeric β-glucosidase (BGLA) was expressed both intra- and extracellularly in a recombinant diploid of Saccharomyces cerevisiae, under limited nutrient conditions. The recombinant enzyme (BGLA15) expressed in the supernatant of a rich medium showed 582 IU/L and 99.4 IU/g dry cell, with p-nitrophenyl-β-d-glucopyranoside as substrate. BGLA15 displayed activity against cello-oligosaccharides with 2–5 glucose monomers, demonstrating that the protein is not specific for cellobiose and that the oligomeric structure is not essential for β-d-1,4-bond hydrolysis. Native β-glucosidase is inhibited almost completely at 160 mM glucose, thus limiting cellobiose hydrolysis. At 200 mM glucose concentration, BGLA15 retained more than 50 % of its maximal activity, and even at 500 mM glucose concentration, more than 30 % of its activity was preserved. Due to these characteristics of BGLA15 activity, recombinant S. cerevisiae is able to utilize cellulosic materials (cello-oligosaccharides) to produce bioethanol.


Cellulomonas flavigena β-Glucosidase Inhibition resistant Saccharomyces cerevisiae 



This work was supported by Consejo Nacional de Ciencia y Tecnología México (CONACYT) (Grant 104333). D. J. Mendoza-Aguayo received a scholarship number 204305 from CONACYT México. María Isabel Pérez- Montfort corrected the English version of the manuscript.


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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • David J. Mendoza-Aguayo
    • 1
  • Héctor M. Poggi-Varaldo
    • 1
  • Jaime García-Mena
    • 2
  • Ana C. Ramos-Valdivia
    • 1
  • Luis M. Salgado
    • 3
  • Mayra de la Torre-Martínez
    • 4
  • Teresa Ponce-Noyola
    • 1
    Email author
  1. 1.Departamento de Biotecnología y BioingenieríaCentro de Investigación y de Estudios Avanzados del Instituto Politécnico NacionalZacatencoMéxico
  2. 2.Departamento de Genética y Biología MolecularCentro de Investigación y de Estudios Avanzados del Instituto Politécnico NacionalZacatencoMéxico
  3. 3.CICATA, Unidad QuerétaroInstituto Politécnico NacionalQuerétaroMéxico
  4. 4.Departamento de Análisis de los AlimentosCentro de Investigación en Alimentación y DesarrolloHermosilloMéxico

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