title: Production of heterologous polygalacturonase I from Aspergillus kawachii in Saccharomyces cerevisiae in batch and fed-batch cultures.
abstract: The pg1 gene from the filamentous fungus Aspergillus kawachii, which codifies for an acid polygalacturonase, was cloned into the pYES2 expression vector, giving rise to the pYES2:pg1ΔI construct. Engineered Saccharomyces cerevisiae, transformed with pYES2:pg1ΔI construct, both expressed and exported an active polygalacturonase with a MW of ~60 kDa and an isoelectric point of 3.7, similar to those reported for the wild-type enzyme. The recombinant enzyme has the ability to hydrolyze polygalacturonic acid at pH 2.5. Heterologous PG1 production was studied under controlled conditions in batch and fed-batch systems. A simultaneous addition of glucose and galactose was found to be the most suitable feeding strategy assayed, resulting in a final PG1 production of 50 U/ml. The production process proposed in this study could be applied for the industrial production of a novel and useful polygalacturonase.
authors.affiliation: Centro de Investigación y Desarrollo en Fermentaciones Industriales, CINDEFI (UNLP; CCT-La Plata, CONICET), 50 y 116 (B1900ASH), La Plata, Argentina.
authors.forename: N L
authors.initials: NL
authors.lastname: Rojas
authors.forename: G E
authors.initials: GE
authors.lastname: Ortiz
authors.forename: D J
authors.initials: DJ
authors.lastname: Baruque
authors.forename: S F
authors.initials: SF
authors.lastname: Cavalitto
authors.forename: P D
authors.initials: PD
authors.lastname: Ghiringhelli
mesh_terms: D001230:Aspergillus / Q000201:enzymology* / Q000235:genetics; D001483:Base Sequence; D061249:Batch Cell Culture Techniques; D005690:Galactose / Q000378:metabolism; D005822:Genetic Vectors; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D010368:Pectins / Q000378:metabolism; D011096:Polygalacturonase / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*
chemical_list: D010368:Pectins; D011096:Polygalacturonase; C003181:polygalacturonic acid; D005690:Galactose