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: Berlin / Heidelberg; Año: 2010

1367-5435

The pg1 gene from the filamentous fungus Aspergillus kawachii, which codifies foran acid polygalacturonase, was cloned into the pYES2 expression vector giving riseto the pYES2:PG1ÄI construct. Engineered Saccharomyces cerevisiae, transformedwith pYES2:PG1ÄI construct, both expressed and exported an activepolygalacturonase with a MW of ~60 kDa and an isoelectric point of 3.7, similar tothose reported for the wild type enzyme. The recombinant enzyme has the ability tohydrolyze polygalacturonic acid at pH 2.5. Heterologous PG1 production wasstudied under controlled conditions in batch and fed-batch systems. A simultaneousaddition of glucose and galactose was found to be the most suitable feedingstrategy assayed, it resulting in a final PG1 production of 50 U/ml. The productionprocess proposed in this study could be applied for the industrial production of anovel and useful polygalacturonase.