2012 ; 22(11):
|Author||Il-Sup Kim, Young-Saeng Kim, Ho-Sung Yoon|
|Affiliation||Advanced Bio-resource Research Center, Kyungpook National University, Daegu 702-701, Korea|
|Title||Glutathione Reductase from Oryza sativa Increases Acquired Tolerance to Abiotic Stresses in a Genetically Modified Saccharomyces cerevisiae Strain|
J. Microbiol. Biotechnol.2012 ; 22(11):
|Abstract||Glutathione reductase (GR, E.C. 188.8.131.52) is an important
enzyme that reduces glutathione disulfide (GSSG) to a
sulfydryl form (GSH) in the presence of an NADPHdependent
system. This is a critical antioxidant mechanism.
Owing to the significance of GR, this enzyme has been
examined in a number of animals, plants, and microbes.
We performed a study to evaluate the molecular properties
of GR (OsGR) from rice (Oryza sativa). To determine
whether heterologous expression of OsGR can reduce the
deleterious effects of unfavorable abiotic conditions, we
constructed a transgenic Saccharomyces cerevisiae strain
expressing the GR gene cloned into the yeast expression
vector p426GPD. OsGR expression was confirmed by a
semiquantitative reverse transcriptase polymerase chain
reaction (semiquantitative RT-PCR) assay, Westernblotting,
and a test for enzyme activity. OsGR expression
increased the ability of the yeast cells to adapt and recover
from H2O2-induced oxidative stress and various stimuli
including heat shock and exposure to menadione, heavy
metals (iron, zinc, copper, and cadmium), sodium dodecyl
sulfate (SDS), ethanol, and sulfuric acid. However, augmented
OsGR expression did not affect the yeast fermentation
capacity owing to reduction of OsGR by multiple factors
produced during the fermentation process. These results
suggest that ectopic OsGR expression conferred acquired
tolerance by improving cellular homeostasis and resistance
against different stresses in the genetically modified yeast
strain, but did not affect fermentation ability.|
|Keywords||Oryza sativa, Glutathione reductase, Abiotic stress, Stress tolerance, Yeast|
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