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.
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Acknowledgments
This research work was supported by CONICET (Consejo Nacional de Ciencia y Tecnología). PDG and SFC are members of the Research Career of CONICET; NLR, DJB and GEO hold a fellowship of CONICET. The authors wish to thank Dr. Donald F. Haggerty, a retired career investigator and native English speaker, for editing the final version of the manuscript.
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Rojas, N.L., Ortiz, G.E., Baruque, D.J. et al. Production of heterologous polygalacturonase I from Aspergillus kawachii in Saccharomyces cerevisiae in batch and fed-batch cultures. J Ind Microbiol Biotechnol 38, 1437–1447 (2011). https://doi.org/10.1007/s10295-010-0929-9
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DOI: https://doi.org/10.1007/s10295-010-0929-9