Effect of nonenzymatic deamidation on the structure stability of Camelus dromedarius α-lactalbumin.

Camelid α-lactalbumin is the only known protein that can undergo nonenzymatic deamidation on two Asn residues. This leads to the generation of a mixture of unusual isoAsp and d-Asp residues that may impact health. The effect of deamidation on camel α-lactalbumin instability was investigated. Circular dichroism showed that the altered protein acquired secondary structure resulting in an increase in α-helix content. In good agreement, the 3D structure of camel α-lactalbumin determined by X-ray crystallography, displayed a short additional α-helix probably induced by deamidation, compared to the human and bovine counterparts. This α-helix was located in the C-terminal region and included residues 101-106. Differential scanning calorimetry together with the susceptibility to thermolysin showed that the deamidation process reinforced the structural stability of the α-lactalbumin at high temperature and its resistance toward proteolysis.

Identification by FT-ICR-MS of Camelus dromedarius α-lactalbumin variants as the result of nonenzymatic deamidation of Asn-16 and Asn-45.

Nonenzymatic deamidation of asparaginyl residues can occur spontaneously under physiological conditions principally when a glycyl residue is at the carboxyl side of Asn and leads to formation of aspartyl and isoaspartyl residues. This modification can change the biological activity of proteins or peptides and trigger an auto-immune response. The α-lactalbumins of members of the Camelidae family are the only of described α-lactalbumins that carry two AsnGly sequences. In the present study, high-resolution mass spectrometry, which enables accurate mass measurement has shown that Asn(16) and Asn(45) underwent a nonenzymatic deamidation, the sequence Asn(45)-Gly(46) being deamidated spontaneously at near-neutral and basic pH and Asn(16)-Gly(17) rather at basic pH. The 16-17 sequence was probably stabilized at near-neutral pH by hydrogen bonds according to the molecular modelisation performed with the camel protein.

Nutritional and hygienic quality of raw milk in the mid-northern region of Algeria: correlations and risk factors.

This paper aims to study the overall quality of raw milk in the mid-northern region of Algeria. The analysis results showed a decrease in the average temperature for the delivery of 1,54°C with P<0.001. However, no significant variation (P>0.05) was observed in almost all the physical and nutritional parameters studied (pH, fat content, and protein content) between M1 and M2. The average contamination by total mesophilic aerobic bacteria (TMAB), coliforms, yeasts, molds, and different pathogens in samples taken at M1 showed significant changes at M2. This was confirmed by the decrease of reduction time of methylene blue (RTMB), about 54%. The variation was described as follows: (P>0.05) for yeasts and (P<0.05) for molds in M1 and M2, (P<0.05) for TMAB in M1, and (P<0.001) for TC, FC, and TMAB in M2. The analysis for the detection of Salmonella spp. showed no contamination in all samples tested, while antibiotic residues were detected in 35% of milks delivered. In conclusion, several risk factors have been identified in this study, namely, the effect of the season and the distance between the farm and the dairy unit.