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: Article

14 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 originalEnglish
Páginas (desde-hasta)507-515
Número de páginas9
PublicaciónFungal Biology
Volumen118
N.º5-6
DOI
EstadoPublished - 1 ene 2014

Huella dactilar

Talaromyces purpurogenus
pectin lyase
Pichia pastoris
Pichia
Penicillium
pectins
enzyme
Enzymes
enzymes
lignocellulose
galacturonic acid
acid
gene
food industry
Complementary DNA
homogeneity
cleavage
methanol
Citrus
Esterification

ASJC Scopus subject areas

  • Ecology, Evolution, Behavior and Systematics
  • Genetics
  • Plant Science
  • Infectious Diseases

Citar esto

Pérez-Fuentes, Claudio ; Cristina Ravanal, María ; Eyzaguirre, Jaime. / Heterologous expression of a Penicillium purpurogenum pectin lyase in Pichia pastoris and its characterization. En: Fungal Biology. 2014 ; Vol. 118, N.º 5-6. pp. 507-515.
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Heterologous expression of a Penicillium purpurogenum pectin lyase in Pichia pastoris and its characterization. / Pérez-Fuentes, Claudio; Cristina Ravanal, María; Eyzaguirre, Jaime.

En: Fungal Biology, Vol. 118, N.º 5-6, 01.01.2014, p. 507-515.

Resultado de la investigación: Article

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AU - Pérez-Fuentes, Claudio

AU - Cristina Ravanal, María

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