Heterologous expression of a Penicillium purpurogenum pectin lyase in Pichia pastoris and its characterization

Claudio Pérez-Fuentes, María Cristina Ravanal, Jaime Eyzaguirre

Resultado de la investigación: Contribución a una revistaArtículo

15 Citas (Scopus)

Resumen

Lignocellulose is the major component of plant cell walls and it represents a great source of renewable organic matter. One of lignocellulose constituents is pectin. Pectin is composed of two basic structures: a 'smooth' region and a 'hairy' region. The 'smooth' region (homogalacturonan) is a linear polymer of galacturonic acid residues with α-(1→4) linkages, substituted by methyl and acetyl residues. The 'hairy' region is more complex, containing xylogalacturonan and rhamnogalacturonans I and II. Among the enzymes which degrade pectin (pectinases) is pectin lyase (E.C. 4.2.2.10). This enzyme acts on highly esterified homogalacturonan, catalysing the cleavage of α-(1→4) glycosidic bonds between methoxylated residues of galacturonic acid by means of β-elimination, with the formation of 4,5-unsaturated products. In this work, the gene and cDNA of a pectin lyase from Penicillium purpurogenum have been sequenced, and the cDNA has been expressed in Pichia pastoris. The gene is 1334pb long, has three introns and codes for a protein of 376 amino acid residues. The recombinant enzyme was purified to homogeneity and characterized. Pectin lyase has a molecular mass of 45kDa as determined by SDS-PAGE. It is active on highly esterified pectin, and decreases 40% the viscosity of pectin with a degree of esterification ≥85%. The enzyme showed no activity on polygalacturonic acid and pectin from citrus fruit 8% esterified. The optimum pH and temperature for the recombinant enzyme are 6.0 and 50°C, respectively, and it is stable up to 50°C when exposed for 3h. A purified pectin lyase may be useful in biotechnological applications such as the food industry where the liberation of toxic methanol in pectin degradation should be avoided.

Idioma originalInglés
Páginas (desde-hasta)507-515
Número de páginas9
PublicaciónFungal Biology
Volumen118
N.º5-6
DOI
EstadoPublicada - 1 ene 2014

Áreas temáticas de ASJC Scopus

  • Ecología, evolución, comportamiento y sistemática
  • Genética
  • Botánica
  • Enfermedades infecciosas

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