Fortification of thermal and structural stability of hemoglobin using choline chloride-based deep eutectic solvents.

Of late, DESs have occupied the centre stage due to their eco-friendly and resource-efficient nature and their low toxicity. In this work, we have investigated the structural and thermal stability of hemoglobin (Hb) in two choline chloride ([Ch]Cl)-based DESs namely urea [Ch]Cl-urea (DES1) and [Ch]Cl-glycerol (Gly); (DES 2). Different biophysical techniques reveal that the presence of DESs facilitates the stability of Hb in a concentration-dependent manner and the extent of stability is more pronounced in [Ch]Cl-Gly as compared to [Ch]Cl-urea. Additionally, for a better understanding of the role of DESs in modulating the thermal and structural stability of Hb, studies have been performed on Hb in the presence of individual constituents of DESs, i.e., [Ch]Cl, urea, and Gly. Altogether, it was observed that the effect on the stability of Hb was by the presence of the DESs rather than their individual constituents. For instance, urea itself is a destabilizing co-solvent for biomolecules. However, the harmful effects of urea were surpassed when a DES is formed in the presence of [Ch]Cl. Therefore, overall, it can be concluded that both DESs can be described as potential non-harmful, green, and promising solvents for enhancing the structural and thermal stability of Hb.

Pea protein-derived tripeptide LRW shows osteoblastic activity on MC3T3-E1 cells via the activation of the Akt/Runx2 pathway.

Osteoporosis is a bone disease affecting more than 2 million people comprising 1 in 3 women and 1 in 5 men in Canada. One possible approach to prevent this disease is to stimulate the activity of osteoblasts (bone-forming cells) using food protein-derived bioactive peptides. In our previous study, an ACE inhibitory tripeptide LRW (Leu-Arg-Trp) was identified from pea protein. This work aims to investigate the effect of tripeptide LRW on promoting osteoblastic activity. The tripeptide LRW treatment (50 µM) in MC3T3-E1 cells increased cell proliferation (4-fold increase) as indicated by BrdU incorporation assay. Moreover, we found that tripeptide LRW stimulated osteoblastic differentiation by increasing the levels of type 1 collagen (COL1A2; 3-fold increase), alkaline phosphatase (ALP; 4-fold increase), and runt-related transcription factor 2 (Runx2; 2-fold increase) and the activation of the protein kinase B (Akt) signaling pathway. Furthermore, tripeptide LRW increased matrix mineralization as evidenced by Alizarin-S red staining and nodule formation, osteoprotegerin levels (OPG; 2-fold increase), and wound healing based on cell migration assay. Overall, pea protein-derived bioactive peptide LRW can positively modulate the activity of osteoblasts probably via the Akt/Runx2 pathway, indicating its potential use for the prevention of osteoporosis.