Friday, 22nd September 2017

Virology and Microbiology

    Yeast enzymes bioengineering to generate bioactive compounds

 

 

2015 Grupo MFdez Lobato 400px
 


María Fernández Lobato

BSciStaff

BPublications

 

Research summary:

We work with microorganisms of biotechnological interest, mainly fungi and yeasts, producers of bioactive compounds. We try to connect the generation of knowledge to the development of biotechnological applications. Basically we focus on the characterization of new enzymes producing bioactive compounds, the analysis of their structural-functional determinants, the operational improvement using molecular biology tools and in obtaining and characterization of new molecules with potential industrial utility. We have patented in different countries the industrial applicability of most proteins characterized and designed methods for their attachment to solid supports.

Fig1 300px   Fig 02 300px
The Xd-INV from Xanthophyllomyces dendrorhous 3D-Structure, a fructofuranosidase producing the prebiotic neokestose. Close-up view of active site including neo-erlose (fructosil maltose).

 

The conformation of the substrates at the Ffase active site. A. Six units of inulin (left) and fructosylnystose (right) molecules represented as spheres. B. Inulin (brown) and fructosylnystose (lime green) moieties in stick representation are superimposed.

 

During the last years we have been characterizing and studying several non-conventional yeast proteins (from genera Xanthophyllomyces, Schwanniomyces, Rhodotorula, etc.) showing glycosyltransferase activity, and applicable in the production of sugars with prebiotic properties. All are glycosylhydrolases (GH) structurally included in family GH32, 31, 13 or 2. Indeed, we have resolved the 3-D structure of the first yeast protein including in family GH32, assigned a function to the beta-sandwich domain that is present in all members of this family and proved that the oligomerization is directly involved in the substrate recognition and specificity. We have improved enzymes and altered their biosynthetic activity using molecular bioengineering tecniques.  More information:

Glicoenz200

   INMARE

 


 

Selected publications:

  • Ramírez-Escudero, M., M. Gimeno-Pérez, B. González, D. Linde, Z. Merdzo, M. Fernández-Lobato* and J. Sanz-Aparicio* (2016) Structural Analysis of β-Fructofuranosidase from Xanthophyllomyces dendrorhous Reveals Unique Features and the Crucial Role of N-glycosylation in Oligomerization and Activity. J. Biol. Chem (doi/10.1074/jbc.M115.708495). *Both corresponding authors.
  • Gimeno-Pérez,M., D. Linde, L. Fernández-Arrojo, F. J. Plou, and M. Fernández Lobato (2015) Heterologous overproduction of β-fructofuranosidase from Xanthophyllomyces dendrorhous, an enzyme producing prebiotic sugars. Appl Microbiol Biotechnol. 98 (8) 3459-3467 (doi: 10.1007/s00253-014-6145-1).
  • de Abreu, M.A., Alvaro-Benito, M., Plou, F.J., Fernández Lobato, M.* and Alcalde, M.* (2013) Synthesis of 6-kestose using a highly efficient β-fructofuranosidase engineered by directed evolution. Adv. Synth. Catal. 355 (9), 1698-1702. * Both corresponding authors. (doi: 10.1002/adsc.201200769).
  • Linde, D., Estévez, M., Plou, F. J. and Fernández Lobato, M. (2012) Analysis of the neofructooligosaccharides production mediated by the extracellular β-fructofuranosidase Xd-INV from Xanthophyllomyces dendrorhous. Bioresour. Technol. 109, 123-130 (doi: 10.1016/j.biortech.2012.01.023)
  • Alvaro-Benito, M., Sainz-Polo, M.A., González, B., Plou, F.J., Fernández-Lobato*, M. and Sanz-Aparicio*, J. (2012) Structural and kinetic insight reveal that the amino acid pair Gln-228/Asn-254 modulates the transfructosylating specificity of Schwanniomyces occidentalis β-fructofuranosidase, and enzyme that produces prebiotics. J. Biol. Chem. 287, 19674-19686. *Both corresponding authors. (doi:10.1074/jbc.M112.355503)

 

Selected patents:

P200402994. New enzyme for the prebiotic production: WO 2006/064078 A1; PCT-ES2005/070177; Europa No 05825223.0. Primer premio a la mejor patente Madrid+d 2008.

P200501875. New fructofuranosydase for prebiotic oligosaccharides obtaining: PCT-ES2006/000435. Europa Nº 06807883.1; USA No. 11/997.233

P200503195. "New fructofuranosydase for the 6-kestose production: PCT-ES2006/000693; Europea No. 06841745-0; USA No. 12/159.164; Japón No. 2008-547993.

P200930001. Biocatalizador inmovilizado basado en alginato para la biotransformación de carbohidratos: PCT-ES2010/070104


 

Selected Doctoral Theses:

  • Patricia Gutiérrez Alonso (2013) Caracterización de dos glicosiltransferasas de oligosacáridos prebióticos de las levaduras Phaffia rhodozyma y Rhodotorula dairenensis. UAM.
  • Miguel Antonio de Abreu Felipe (2011) Studies aimed at improving the funtionality of non-conventional yeast enzymes able to synthesize prebiotic oligosaccarides. UAM. European Program
  • Miguel Álvaro Benito (2011) The study of β-fructofuranosidase from Schwanniomyces occidentalis reveals new functional elements in the family GH32 of glycosyltransferases and an unconventional genetic code use in this yeast. UAM. European Program
  • María Dolores Linde López (2010) Caracterización Bioquímica, molecular y estructural de una β-fructofuranosidasa con capacidad transferasa de la levadura Phaffia rhodozyma aplicable a la producción de oligosacáridos prebióticos. UAM

 

Images gallery:

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