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Efficient expression of a Paenibacillus barcinonensis endoglucanase in Saccharomyces cerevisiae

  • Original Paper
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Journal of Industrial Microbiology & Biotechnology

Abstract

The endoglucanase coded by celA (GenBank Access No. Y12512) from Paenibacillus barcinonensis, an enzyme with good characteristics for application on paper manufacture from agricultural fibers, was expressed in Saccharomyces cerevisiae by using different domains of the cell wall protein Pir4 as translational fusion partners, to achieve either secretion or cell wall retention of the recombinant enzyme. Given the presence of five potential N-glycosylation sites in the amino acid sequence coded by celA, the effect of glycosylation on the enzymatic activity of the recombinant enzyme was investigated by expressing the recombinant fusion proteins in both, standard and glycosylation-deficient strains of S. cerevisiae. Correct targeting of the recombinant fusion proteins was confirmed by Western immunoblot using Pir-specific antibodies, while enzymatic activity on carboxymethyl cellulose was demonstrated on plate assays, zymographic analysis and colorimetric assays. Hyperglycosylation of the enzyme when expressed in the standard strain of S. cerevisiae did not affect activity, and values of 1.2 U/ml were obtained in growth medium supernatants in ordinary batch cultures after 24 h. These values compare quite favorably with those described for other recombinant endoglucanases expressed in S. cerevisiae. This is one of the few reports describing the expression of Bacillus cellulases in S. cerevisiae, since yeast expressed recombinant cellulases have been mostly of fungal origin. It is also the first report of the yeast expression of this particular endoglucanase.

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Acknowledgments

This work was supported by grant ISCIII2006-PI0731 from the Ministerio de Sanidad/Instituto de la Salud Carlos III (Spain). María Mormeneo was a recipient of a pre-doctoral grant from the Programa Nacional de Formación de Profesorado Universitario del Ministerio de Educación y Ciencia.

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  1. Unidad de Microbiología, Facultad de Farmacia, Universidad De Valencia, Avda. Vicente Andrés Estelles s/n, 46100, Burjassot, Valencia, Spain

    María Mormeneo & Jesús Zueco

  2. Departamento de Microbiología, Facultad de Biología, Universidad de Barcelona, Avenida Diagonal 645, 08028, Barcelona, Spain

    FI Javier Pastor

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  1. María Mormeneo
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  2. FI Javier Pastor
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Correspondence to Jesús Zueco.

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Mormeneo, M., Pastor, F.J. & Zueco, J. Efficient expression of a Paenibacillus barcinonensis endoglucanase in Saccharomyces cerevisiae . J Ind Microbiol Biotechnol 39, 115–123 (2012). https://doi.org/10.1007/s10295-011-1006-8

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