Muhammad Azrul Hafiz Bin Kamaruddin

Saccharomyces Cerevisiae As A Model To Study Oxidative Stress And The Role Of Vitamin E
As Antioxidants. (A Review)

INTRODUCTION

Saccharomyces cerevisiae is one of eukaryotic organisms that is used to study eukaryotic

system especially in higher eukaryotes. Their property single cells and the ability to reproduce
asexually via budding making them easier to be use as a model study in answering biological
questions. This yeast have served as an important organism since the 1930s in understanding the
biochemical properties and activities as well as helped in developing genomics tools as they are
not causative of any disease, easy to be manipulated using genetic tools and possess a simple
genetic system (Goffeau et al., 1996; Forsburg, 2001). Karathia et al. (2011) describes that
S.cerevisiae are the appropriate model to study pathologies in human as they possess similar
biological processes that can serve as a substitute. In addition, S. cerevisiae possess similar set of
proteins that regulate and execute biological processes as compared to humans.

Vitamin E is the common name for the four tocopherols and four cortineols, namely -,
-, -, and -, are only obtainable by consumption in human. Through studies, it is found out that
only -tocopherol meets the human vitamin E requirement. In eukaryotes, these -tocopherol
exists in the membrane and widely known to affect the aging effect. These antioxidant acts in a
regulating system with other antioxidants and function especially on regulating lipid
peroxidation. (Sattler et al. 2006)

This review will centralize on S. cerevisiae as a model to study oxidative stress and the
role of vitamin E as antioxidant.

OXIDATIVE STRESS ON S. CEREVISIAE

Reactive oxygen species (ROS) exist naturally in every aerobic cells through the
metabolic activities that were carried out in mitochondria. Some of these ROS include
superoxide anions, hydroxyl radicals, singlet oxygen and nitric oxide. These ROS is believed to
be the cause of oxidative stress a state where imbalance in the redox couples occurs. It is
believed that the oxidative stress is causing many pathological condition in humans such as
atherosclerosis, neurodegenerative disease and even cancer (Gutteridge 1993 Hybertson et al
2011 Lu et al 2010 ). Halliwell et al. (1993) describe that intermediates of oxidative stress may
target the membrane bilayer and cause the polyunsaturated fatty acids to undergone lipid
peroxidation. This condition will trigger the reaction with lipid that yield a lipid radical and a
lipid hydroperoxide (LOOH). These products will breakdown into multiple by-products that are
highly reactive.

In S. cerevisiae, multiple mechanisms are employed by the cells to protect against

oxidative damages. Some of these mechanisms include enzymes such as catalase, superoxide
dismutase and gluthathione peroxidase as well as antioxidants such as gluthathione (GSH) and
vitamins C and E (Jamieson 1998). In respond to oxidative stress two system have been
observed to play an impotant role, that is the gluthathione/glutaredoxin and thioredoxins. Both
glutharedoxins and thioredoxins have been identified as the heat-stable oxidoreductase and are
involve in catalytic cycle with metabolic enzymes that function in protein folding and regulation,
oxidatively damaged proteins and sulphur metabolism (Demple, 1998; Holmgren 1989).

I) Glutathione/ glutaredoxin system respond to oxidative stress.

Grant & Dawes (1996) explain that the reduced glutathione levels are regulated by a
series of complex reactions that involve the enzymes of biosynthesizing such as
-gutamylcysteine sythetase(GSH1) and glutathione sythetase(GSH2) as well as the redox
regulation of glutathione reductase (GLR1).

Hydroperoxides are broken down by glutathione peroxidase, from the three genes
(GPX1, GPX2 and GPX3) that have similar homology to human glutathione peroxidases (Inoue
et al. 1996). This peroxidase is identified to play a role in oxidative stress when mutants that lack
the ability to synthesis Gpx3 are prone to hydrogen peroxide whereas mutant of GPX1 are unable
to grow under stress condition. Deffieu et al. (2009) explained that in yeast, mitophagy (in
mitochondria) is strictly mediated by glutathione. ROS that are constantly produced need to be
regulated and therefore, the concentration of glutathione need to be maintained by the cells. This
continuous maintenance cause GSH to be involved in many biological activity including
mitophagy.

The glutaredoxin system on the other hand is encoded by two genes designated as GRX1
and GRX2. Through multiple experiments, mutants of both genes lack oxidoreductase activity,
GRX1 mutants are sensitive towards oxidative stress by superoxide anions while GRX2 mutants
are sensitive towards hydrogen peroxide. It is believed that these genes are responsible in
detoxifying ROS-mediated damage in stress conditions. (Luikenhuis et al. 1997). Luikenhuis
explained that the glutaredoxin system does not affect the GSH levels but are able to affect the
thioredoxin system.

II) Thioredoxin system against oxidative stress.

Thioredoxin peroxidase was first characterized as thiol-specific antioxidant (TS) that

protected cells against a thiol-containing oxidation system.Tsa1 and Tsa2 have shown the
capability to possess peroxidase activity against hydrogen peroxide and alkyl peroxides. Tsa1
seems to prefer hydrogen peroxides whereas Tsa2 show the capability to reduces alkyl
hyperperoxides. This is proven in a test using mutant strains of the yeast that was conducted by
Chae et al. (1993).

Under stress condition, the cells upregulate the synthesis of several molecules to protect
itself against oxidative damage. These molecules include enzymes from GSH/glutaredoxin
pathway as well as the thioredoxin systems.

VITAMIN E AS ANTIOXIDANT

All eight vitamin E molecules are antioxidant lipophilic molecules and are attributed by
their hydrophobic tail of a saturated phytyl chain in tocopherols and unsaturated phytyl chain in
tocotrienols. The antioxidant attribute is due to the presence of the chromanol group (Niki 1987).
In humans, where -tocopherol is preferred, they act as a peroxyl scavenger that terminates chain
reations (Tappel, 1962). -tocopherol have a higher H-atom-donating ability by having a greater
ring methyl substitutions. Besides that -tocopherol have been reported to be able to regulate the
expression of lipoproteins receptors, scavenger receptor BI and its homolog CD36 in oxidative
stress response (Devaraj et al. 2001; Kolleck et al. 2000). This is done by the -tocopherol
protecting the long polyunsaturated fatty acid chains and maintain their concentrations for
important signaling events. These fatty acids are prone to lipid peroxidation by ROS and the state
where vitamin E are deficient, tissue areas may be harm.

This is proven in several test where rabbits that lack vitamin E in their diets have reduced
antioxidant system. The testosterone propionate levels that are increased cause ROS to be
produced at a higher levels during the steroidogenesis. The increase in ROS in the tissues cause a
rapid reduction vitamin E. This show that vitamin E is consumed when the level of ROS is high
(Aydilek et al. 2004). Combs & Scott (1974) observed that the level of vitamin E in chicks are
greatly reduced when an increase in peroxidation in cells is triggered. This observation is also
shared by Soondergard & Dam (1966) where animals with various pathologies due to presence
of rancid fats in tissues are cured when they are fed with foods that have high concentration of
tocopherols. This show that -tocopherol act as a lipid-soluble antioxidant.

Bioactive lipids are important in signaling molecules and they are prone to oxidative
activity of ROS. Presence of vitamin E maintain the integrity and protect them from oxidative
damages.

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