Oral presentation
34
BIOLOGICAL ACTIVITY SCREENING AND FRACTIONATION
OF Naja Naja oxiana VENOM
N.R. Mukhamedov, M.B. Kayumov, Z.Zh. Mirakhmetova, Sh.Ya. Mirzaakhmedov
Institute of Bioorganic Chemistry, 100125, Tashkent, Mirzo Ulugbek street 83.
e mail: mirzaakhmedov@mail.ru
Naja naja oxiana
also called the Central Asian cobra
is an endemic species of
venomous snake in the family
Elapidae
.
Its venom contains mainly peptides and
proteins (80-90%). Drugs produced from snake venom that
are already licensed or in
development show that unpleasant bioactivity may be converted into a therapeutic for
the correct condition. Though some drugs were discovered in the cobra venom, there is
a high possibility of finding new ones relying on the rich chemical constituent of the
object.
Taking into
account the aforesaid opinion, we aimed to separate and isolate novel
biologically active low molecular weight peptides from
Naja oxiana
venom.
To reach
the aim, the following tasks were conducted: 1) biological activity screening of the
venom; 2) ultrafiltration of the venom; 3) comparative analysis of the fractions obtained
from the ultrafiltration by HPLC.
For the screening of the biological activity of the venom,
it was dissolved in water
and tested for antioxidant activity and inhibition of more than 10 types of kits
corresponding to several diseases like cancer, SARS, HIV, etc.
According to the test
results,
Naja naja oxiana
snake venom showed the following 3 types of biological
activities at a concentration of 0.1 mg/ml: (1) on HIV-RT (AIDS) – 100% inhibition; (2)
p300 (cancer) enzyme – 83% inhibition; (3) the antiradical
activity of snake venom was
80% at the initial time and decreased to 67% at 30 minutes.
Venom was dissolved in 0.1 M ammonium acetate pH 4 buffer to overcome
proteolytic hydrolysis of the components. Ultrafiltration was carried out using
ultrafiltration membranes 10 kDa and 3 kDa sizes. >10 kDa, <10->3 kDa, and <3 kDa
fractions of the venom were obtained as a result. The fractions were lyophilized and
stored at -20
o
C for further experiments.
HPLC analysis of the venom and its fractions obtained
after ultrafiltration was
carried out using the C-18 column in the gradient mode of MeCN against 0.1 % TFA.
HPLC profiles of all samples were analyzed using Origin software. The HPLC
chromatogram of the venom consisted of two parts: 1) the hydrophilic part which was
not adsorbed well in the column and was detected before 5 minutes; 2) the hydrophobic
3>
Dostları ilə paylaş: