Preparation, characterization and bioavailability studies of Tegillarca granosa hemoglobin and its glycosylated products.

Iron deficiency anemia (IDA) is a common micronutrient deficiency. Tegillarca granosa (T. granosa) is a good source of iron due to its high content of hemoglobin. The present study aimed to determine the effects of glycosylation on structure, physicochemical characteristics and iron bioavailability of hemoglobin. Using Box-Behnken design and response surface methodology, the optimal conditions for hemoglobin-chitosan glycosylation were obtained: 61.8 °C, pH 6.3, hemoglobin/chitosan mass ratio of 4.3 and reaction time of 15 min. The formation of hemoglobin-chitosan conjugates was verified by SDS-PAGE and fluorescence spectroscopy. The surface hydrophobicity of hemoglobin was reduced by 20.90-65.05 % after glycosylation, along with the observations of elevated water-holding capacity, likely owing to the introduction of hydrophilic groups. Antioxidant capacity of glycosylated products (0.41-0.66 μM Trolox/mg protein) was markedly greater than that of original protein (0.06 μM Trolox/mg protein) due to the formation of brown polymers with antioxidant activity. In addition, glycosylation improved in vitro digestibility of hemoglobin by 41.15-69.09 %, which could be attributed to less β-sheet in secondary structures. Moreover, hemoglobin (324.38 ng/mg) exhibited better iron absorption than FeSO4 (121.63 ng/mg), with the value being further enhanced by glycosylation (442.73 ng/mg), which may be due to the improved protein digestibility and iron-chelating capacity.