Extraction and biochemical characterization of sulphated
glycosaminoglycans from chicken keel cartilage
Humaira Majeed Khan1, Muhammad Ashraf2,
Abu Saeed Hashmi3, Mansur-ud-Din Ahmad4 and
Aftab Ahmad Anjum5
1Lahore
College for Women University, Jail Road, Lahore; 2Department
of Pharmacology & Toxicology; 3Institute of Biochemistry
& Biotechnology; 4Department of Epidemiology & Public
Health and 5Department of Microbiology, University of
Veterinary and Animal Sciences, Lahore-54000, Pakistan *Corresponding author:mansuruddin@uvas.edu.pk
Abstract
The present study was conducted to explore the
potential and cheaper source of major and abundantly found sulphated
Glycosaminoglycans in chicken keel cartilage. Chicken is comparatively readily
accessible to all the communities of Pakistan and its cartilages are the rich
source of sulphated GAGs. The GAGs were extracted from prewashed and ground keel
cartilages (n=3) of chicken using 3 M
MgCl2, dialyzed, digested with papain, precipitated with three
volumes of ethanol, and finally lyophilized to dry powder. The dry products were
used for proximate analysis (carbohydrates
65.49±0.10, crude protein
12.82±0.26, ash 11.12±.56,
moisture 9.88±0.32 and fat
0.69±0.14%). Dimethylmethylene
blue binding (DMMB) assay was performed to determine the quantity of total GAGs
in each group of product and protein contents were estimated by Bradford method.
Identification of extracted samples of GAGs was performed with FTIR spectrometer
using KBr disc and purity of the samples was determined by SDS-PAGE. Quantity of
total GAGs in extracted samples was 70.77±2.27% and estimated amount of protein
was 4.64±0.29%. FTIR spectra of
standard and samples of CS showed identical and characteristic peaks in finger
print region. Finger print region revealed the presence of C-O-S, S=O, -COO,
-C-C, R-SO2–R, -CONH2 and R-SO2-NH2
molecules. SDS-PAGE analysis revealed the presence of 77.8 and 50.5 kDa proteins
in all extracted samples of GAGs. It can be concluded that chicken keel
cartilage is the potential and cheap source of GAGs. Analysis by SDS-PAGE
revealed that most of the non-collagen protein can be removed by three volumes
of solvent extraction and FTIR is an advance technique for identification of
GAGs in mid infrared region (400-4000 cm-1).