PRODUCTION OF HETEROLOGOUS POLYGALACTURONASE I FROM ASPERGILLUS KAWACHII IN SACCHAROMYCES CEREVISIAE IN BATCH AND FED-BATCH CULTURES

ROJAS, NATALIA LORENA; ORTIZ, GASTON EZEQUIEL; BARUQUE, DIEGO JORGE; CAVALITTO, SEBASTIÁN FERNANDO; GHIRINGHELLI, PABLO DANIEL

JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY

SPRINGER HEIDELBERG

Lugar: Heidelberg; Año: 2011 vol. 38 p. 1437 - 1447

1367-5435

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, it 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.