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European Journal of Applied Sciences – Vol. 9, No. 6
Publication Date: December 25, 2021
DOI:10.14738/aivp.96.11207. Cakmak, M., Ozer, D., Karatas, F., & Saydam, S. (2021). Combine Effect of Vitamin C and venlafaxine on the Amino Acid Content of
Saccharomyces cerevisiae. European Journal of Applied Sciences, 9(6). 137-153.
Services for Science and Education – United Kingdom
Combine Effect of Vitamin C and venlafaxine on the Amino Acid
Content of Saccharomyces cerevisiae
Meltem ÇAKMAK
Department of Chemical Engineering
Faculty of Engineering, Firat University, 23119 Elazig
Dursun ÖZER
Department of Chemical Engineering
Faculty of Engineering, Firat University, 23119 Elazig
Fikret KARATAS
Faculty of Science, Department of Chemistry
Fırat University, Elazig, Turkey
Sinan SAYDAM
Faculty of Science, Department of Chemistry
Fırat University, Elazig, Turkey
ABSTRACT
In this study, Saccharomyces cerevisiae (NRLLY-12632) was grown in YPD medium
containing different concentrations of venlafaxine, (RS)-1-[2-dimethylamino-1-(4-
methoxyphenyl)-ethyl] cyclohexanol. To counteract the effect of venlafaxine,
vitamin C were added to the growth medium of S. cerevisiae, and amino acids
content were investigated by HPLC. It was observed that, the amount of amino acids
in S. cerevisiae grown in YPD medium, containing 100, 200, 300, 400 and 500 ppm
venlafaxine were compared with the control, the amounts of alanine, arginine,
asparagine, aspartic acid, cysteine, glycine, histidine, isoleucine, leucine, lysine,
phenylalanine, proline, threonine, tyrosine and valine increased (p<0.05), glutamic
acid, glutamine, methionine, serine and tryptophan levels were decreased p<0.05).
With the addition of vitamin C to the growth medium containing venlafaxine, all the
amino acids concentrations get close to the control group. From these findings, it
can be said that the negative effect of venlafaxine on S. cerevisiae is reduced by the
addition of vitamin C to the medium.
Keywords: S. cerevisiae, Venlafaxine, Vitamin C, Amino acid.
INTRODUCTION
Although there are millions of living species in nature, there are common structures and
characters among living things in this diversity. In addition, many genes that humans have,
microorganisms also contain some genes that humans do not have. The term microorganism is
used to describe a wide range of organisms including yeast, bacteria, fungi, algae and even
protozoa. Yeasts are single-celled eukaryotic organisms that are widely found in nature, have
high adaptability, take part in the production of various foods, reproduce by budding or division
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European Journal of Applied Sciences (EJAS) Vol. 9, Issue 6, December-2021
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[1]. In addition to their probiotic properties, yeasts also show antioxidant activity thanks to the
various components they produce (such as superoxide dismutase, catalase, carotenoids,
resveratrol, beta-glucans). Yeasts (especially Saccharomyces cerevisiae) are the most
important group among biotechnological organisms. Yeasts are also important for other
industries (biofuel, single cell protein, industrial enzyme, secondary metabolites with economic
value) and scientific studies (eukaryotic biology and study of human diseases), mainly in the
field of food. The annual production of S. cerevisiae yeast worldwide exceeds that of other
biotechnological microorganisms combined [2]. Yeasts are used in many industries because of
their use of cheap renewable resources as carbon and energy sources to reproduce, which
ability to reproduce at a high rate in a given time, easily changeable properties and low cost [3].
Depression, a psychiatric disorder, is the most common complex disease in the world, leading
to serious dysfunctions [4] which is a complex illness with symptoms such as irritability,
insomnia, fatigue, agitation, psychomotor changes, guilt, and self-devaluation. The most
commonly used method in the treatment of this disease is the use of antidepressants [5]. Today,
the use of these drugs is increasing rapidly. Side effects associated with antidepressants such
as headache, nausea or vomiting, agitation, sedation, insomnia and sexual dysfunction and
akathisia have been reported [6]. The World Health Organization predicts that depression will
rank second among the diseases that cause death due to stress and complications related to the
cardiovascular system. It is reported that approximately 21% of the population worldwide is
affected by depression [7]. Pharmacological methods and psychotherapies have proven to be
effective in the treatment of depression. Due to, time, and economic reasons, the most
commonly used method of treating depression is the prescription of antidepressant drugs [8].
Antidepressants act as enzyme, receptor inhibitors and reuptake inhibitors. Antidepressant
drugs used in the clinic work by directly or indirectly increasing the effects of dopamine,
norepinephrine or serotonin in the brain. Generally, antidepressant drugs are classified under
eight groups [9]. Like many drugs, antidepressants also cause oxidative stress by affecting cell
metabolism. Radicals formed due to oxidative stress can affect many parameters including cell
membrane, proteins, amino acids, glutathione, malondialdehyde, total oxidant and antioxidant
capacity.
In order to reduce the oxidative effect of antidepressant, it has been investigated by adding
substances such as vitamins A, E and C, which are known for their antioxidant properties.
Vitamin C is effective in the release of some hormones in case of stress and has a strong
antioxidant effect. Vitamin C has a role in tissue repair, protein formation, inactivation of toxic
metals and protection of vitamins, and oxidation of DNA [10].
Amino acids are not only the building block of proteins, but also free amino acids are the
potential energy-related metabolites and secondary metabolites. Considering these features,
amino acids are important substances that provide cell renewal [11], and involved in
neurotransmitter and biosynthesis processes in biological systems. For adequate production of
protein in the body, it is necessary to take essential amino acids by diet. Depending on the
functions of organs in the body, concentration of amino acid can vary. Amino acid metabolism
changes under various physiological and pathological conditions, leading to change in whole
body homeostasis. In recent years, the use of amino acid profiling in medical science in
pathological studies and clinical diagnosis has increased the interest on amino acids [12].
Differences in amino acid levels have been observed in various diseases compared to healthy
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Cakmak, M., Ozer, D., Karatas, F., & Saydam, S. (2021). Combine Effect of Vitamin C and venlafaxine on the Amino Acid Content of Saccharomyces
cerevisiae. European Journal of Applied Sciences, 9(6). 137-153.
URL: http://dx.doi.org/10.14738/aivp.96.11207
controls. It has also been reported that amino acids can be used in the follow-up of the course
of some diseases [13] and that significant changes in the amino acid profile can be used for early
diagnosis in various types of cancer [13, 14].
S. cerevisiae is a eukaryote, and has complex intracellular structures of animals and plants, and
sequences of the yeast and human genomes are 23% in common. Since microorganisms can be
reproduced easily and quickly which have many common features with other living things, it
provides a great advantage in terms of medical research. For that reasons, S. cerevisiae was
preferred in the study to investigate the effect of venlafaxine which is in the group of Serotonin- Norepinephrine Reuptake Inhibitors (SNRI) whit the chemical formula of (RS)-1-[2-
dimethylamino-1-(4-methoxyphenyl)-ethyl] cyclohexanol chloride together with vitamin C to
counteract on the amino acids contents of S. cerevisiae
MATERIALS AND METHODS
Material
Saccharomyces cerevisiae (NRLLY-12632) grown in YPD medium (10.0 g peptone, 5.0 g yeast
extract, 10.0 g Dextrose per liter) was used. A stock solution of 1000 ppm venlafaxine chloride
was prepared. 500 ppm Vitamin C stock solution was freshly prepared and used.
Microorganism production was carried out in 250 mL flasks containing 50 mL broth. The
following groups were studied,
1. Control: S. cerevisiae was added to sterile YPD medium.
2. Venlafaxine group: The microorganism was produced by adding desired amounts of
venlafaxine stock solution to the control, (100, 200, 300, 400 and 500 ppm).
3. Vitamin C group: The microorganism was reproduced by adding the required amount of
vitamin C stock solution to the venlafaxine group according to the desired medium
concentration (10, 25, 50 and 75 ppm).
After the inoculation, it was incubated at 30 °C with 150 rpm, for 72 hours in an orbital shaker
(Selecta Rotabit). At the end of the incubation period, the concentration of S. cerevisiae was
determined by measuring the absorbance at 600 nm with UV-Visible spectrophotometer
(CHEBIOS s.r.l.). Then growth medium centrifuged at 10 oC with 8000 rpm, for 10 minutes,
(Nüve NF 800 R) the precipitated S. cerevisiae were washed twice with distilled water, and
centrifuged again and used in further processing.
DETERMINATION OF AMINO ACIDS
Hydrolysis
A certain weight of microorganism taken into a glass tube and 5.0 mL 6.0 N HCl was added and
vortexed thoroughly then, samples were kept at 110 °C for 24 hours to break the peptide bonds
[15]. Than the samples cooled to room temperature, filtered and the filtrate volume was
completed to 15 mL with distilled water.
Derivatization
Standard amino acid solutions were prepared using 0.10 N HCl at different concentrations
between 1.0 to 5.0 μg/mL. 50 μL Standard amino acid solutions or hydrolysed samples
transferred to 5.0 mL glass tubes and dried under vacuum at 65 °C. Then 50 μL of reagent 1 [(2: