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Glycerol is a well-known product with a large variety of applications in processes and products. Bio-diesel industry generates
amounts of glycerol that largely exceed their potential applications. Nevertheless, the crude glycerol does not substitute
for glycerin from petrochemical origin since the refining process requires large-scale complex technology and therefore prices
do not compete. In consequence, crude glycerol, in opposition to glycerin, is poorly utilized charging the biodiesel industry
with a high waste-disposal cost. In this context large amounts/low cost crude glycerol became an interesting carbon source for
the biotechnology industry, but this depends on the use of amenable microorganisms with recognized industrial applications.
Saccharomyces cerevisiae was genetically engineered aiming to obtain strains that consumed crude glycerol efficiently. For
this purpose, several proteins were overexpressed and freed of glucose repression and other tight regulation mechanisms
combining several construction strategies. These included proteins from the glycerol consumption pathway, the glycerol plasma
membrane high affinity transporter (Stl1p) and kinase (Gut1p) and other proteins indirectly influencing ethanol production
flux, the thiamine plasma membrane permease (Thi10p) and pyrophosphokinase (Thi80p) and Pdc2, a transcription factor
for thiamine-regulated genes required for expression of the two pyruvate decarboxylase isoforms. Engineered strains were
subsequently challenged with biodiesel industry crude glycerol-based culture conditions for proof of concept.