Organizing co mmittee

NOVEL ANTIOXIDANT PEPTIDES FROM THE CAMEL
MILK HYDROLYSATES 
 
U.J. Ishimov, Asrorov A.M, J.F. Ziyavitdinov
 
Institute of Bioorganic Chemistry, Academy of Sciences Republic of Uzbekistan, 
Tashkent
e-mail: [e-mail]  
 
Milk peptides have strong biological activities. Camel milk protein composition is 
different from those in cow, buffalo, sheep, goats, donkey and mare based on casein and 
raw protein proportion. In the present study, camel milk proteins were hydrolyzed using 
trypsin. The digested camel milk samples were then fractionated and further their 
antioxidant activities were determined using 2,2-diphenyl-1- picrylhydrazyl (DPPH) 
and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid (ABTS
+
). Antioxidant 
activity test guided-fractionation allowed isolation of the three antioxidant peptides 
using ultrafiltration (3000 Da membrane), gel filtration chromatography, and RP-HPLC. 
Camel milk was defatted by centrifugation at 10 000 rpm for 15 min using a high-
speed centrifuge. Skimmed milk protein was hydrolyzed with trypsin enzyme under 
optimal conditions for 4 hours. The yield of isolated peptides made 82%. Camel milk 
peptides were fractionated and purified using a C18 Agilent column (ZORBAX 300SB-
C18 25.0 cm × 9.4 mm, 5 μm; USA). Elution was monitored with an ultraviolet (UV) 
detector at 224 nm. The amino acid sequence and molecular weight of the separated 
peptide fractions were determined on an Agilent Technologies 6530B Chip-HPLC-Q-
TOF mass spectrometer. The obtained mass spectra were interpreted using Mass COT 
and Spectrum Mill programs. As a result of chromato-mass spectrometric studies, the 
following fragment peptides were identified: F-1 - RLDGQGRPRVWLGR, F-2 - 
TPDNIDIWLGGIAEPQVKR and F-3 - VAYSDDGENWTEYRDQGAVEGK with 
MW 1665.94, 2122.13 and 2489.09 Da, respectively. The amino acid sequence search 
in databases showed that the peptides were novel. 
The DPPH and ABTS
+
scavenging activities of three individual peptides (F-1, F-2, 
and F-3) were determined. IC
50
values on DPPH were 1.9 for F-1, 1.2 for F-2, 0.6 
mg/mL for F-3. Results on ABTS
+
made 2.4, 1.8 and 0.9 mg/mL respectively. F-3 
exhibited higher DPPH and ABTS scavenging activity than other fractions.