Characteristics of Saccharomyces cerevisiae gal1Δ and gal1Δ hxk2Δ mutants expressing recombinant proteins from the GAL promoter

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Title
Characteristics of Saccharomyces cerevisiae gal1Δ and gal1Δ hxk2Δ mutants expressing recombinant proteins from the GAL promoter
Author(s)
Hyun Ah Kang; W K Kang; S M Go; A Rezaee; S H Krishna; Sang Ki Rhee; J Y Kim
Bibliographic Citation
Biotechnology and Bioengineering, vol. 89, no. 6, pp. 619-629
Publication Year
2005
Abstract
Galactose can be used not only as an inducer of the GAL promoters, but also as a carbon source by Saccharomyces cerevisiae, which makes recombinant fermentation processes that use GAL promoters complicated and expensive. To overcome this problem during the cultivation of the recombinant strain expressing human serum albumin (HSA) from the GAL10 promoter, a gal1Δ mutant strain was constructed and its induction kinetics investigated. As expected, the gal1Δ strain did not use galactose, and showed high levels of HSA expression, even at extremely low galactose concentrations (0.05-0.1 g/L). However, the gal1Δ strain produced much more ethanol, in a complex medium containing glucose, than the GAL1 strain. To improve the physiological properties of the gal1Δ mutant strain as a host for heterologous protein production, a null mutation of either MIG1 or HXK2 was introduced into the gal1Δ mutant strain, generating gal1Δmig1Δ and gal1Δhxk2Δ double strains. The gal1Δhxk2Δ strain showed a decreased rate of ethanol synthesis, with an accelerated rate of ethanol consumption, compared to the gal1Δ strain, whereas the gal1Δmig1Δ strain showed similar patterns to the gal1Δ strain. Furthermore, the gal1Δhxk2Δ strain secreted much more recombinant proteins (HSA and HSA fusion proteins) than the other strains. The results suggest that the gal1Δhxk2Δ strain would be useful for the large-scale production of heterologous proteins from the GAL10 promoter in S. cerevisiae.
Keyword
GAL promoterGAL1HXK2Recombinant proteinSaccharomyces cerevisiae
ISSN
0006-3592
Publisher
Wiley
DOI
http://dx.doi.org/10.1002/bit.20240
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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