Journal of Biotechnology & Biomaterials
Open Access

Abstract

More than 100 heterologous proteins from various phylogenetic origins have been successfully produced in Yarrowia lipolytica yeast expression system, which offers reliable performances in glycosylation, folding and secretion of complex proteins. We present here a short review of the major tools developed in this yeast, with focus on vectors for targeted monocopy integration (YLEX expression kit, Yeastern Biotech Co, Taiwan), a system particularly adapted to genetic engineering of heterologous proteins. In order to examplify the various applications of Yarrowia expression system, a number of current collaborative works on whole-cell biocatalysis, genetic engineering of secreted enzymes, and surface-display of heterologous proteins (arming yeasts) will be briefly resumed. Some applications, like high throughput screening, require improving Yarrowia transformation methods for higher efficiency, without sacrificing the high percentage of correctly targeted integrations. For that purpose, we have developed a sensor system for homologous integration, based on E. coli lacZ gene expression, which allowed us to evaluate some chemicals increasing transformation efficiency for maintenance of a high percentage of correct integration. Dithiothreitol was found to be not only more efficient than dimethyl-sulfoxide in increasing transformation efficiency, but also compatible with homologous integration, which was not the case of the latter that promoted out-of-site integration. An optimized transformation protocol using dithiothreitol has been developed, which ensures very high transformation efficiency.

Biography

Catherine Madzak has been a student at Ecole Normale Sup?rieure, one of the main French governmental schools of higher education. She obtained her Ph.D. in Microbiology at the Cancer Institute of Villejuif. She is now a senior research associate at INRA, the French National Institute for Agricultural Research. Her main research subject is to develop an expression system for heterologous proteins in the yeast Yarrowia lipolytica, with particular emphasis on genetic engineering of enzymes. She is author of more than 40 publications in international journals, five book chapters, and inventor of a patent on a recombinant promoter.