Characterization of Protein Nanoparticles and their Binding to Textiles
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Abstract‘Smiling in your brother’s face is an act of charity’; said prophet Muhammad (ALLAH’s peace and blessings be with him). Amelogenin (Amg) is one of the most important proteins that are responsible for perfect tooth formation giving us that beautiful smile. Amelogenin has multifunctional characteristics like wound healing, cells proliferation induction, anti-inflammatory, anti-infectious and signaling properties in living organisms. Its high tendency to aggregate and self-assemble into nanospheric structures is employed biologically to control important physiological processes, like bio-mineralization, and induction of specific cellular responses. Having a distinct solubility profile, being soluble at high acidic and high alkaline pH, while it aggregates at physiologic pH in the form of nanospheres, Amg can be purified in a fairly simple one step acid-heat treatment purification method. In the current study, Amg nanospheres were examined for their binding capability to cellulosic materials. Amg wild type (rh174) and Amg fused with cellulose-binding domain (rh174_CBD), were successfully produced in E. coli and purified to homogeneity in high levels (~ 95% of total protein), (~ 107,7 and 195,25 mg/l culture) for purified rh174 and rh174_CBD, respectively. The purified proteins were characterized by (SDS-PAGE) and showed homogeneity indicating efficiency of the purification step. DLS was also performed at pH 10 with 0.5 mg/ml and resulted in highly homogenous, successfully assembled, uniform nanospheres with ~ 35 ± 2 nm hydrodynamic radius, which reflects their stability under these conditions. Then different Amg variants were tested for their binding capability to cellulose. The results showed superior affinity of rh174_CBD to cellulose despite the highly alkaline conditions. The immobilization yield of rh174_CBD reached up to ~ 99.4 % and a binding capacity up to 7 mg/g cellulose.
‘Smiling in your brother’s face is an act of charity’; said prophet Muhammad (ALLAH’s peace and blessings be with him). Scientists have approved how important a smile is, as a personal characteristic, in social communication for human being. Proteins are a group of molecules that function as constructive workers, and restrictive managers of any physiological process in any living micro/ macro organism. Amelogenin (Amg) is one of the most important proteins that are responsible for perfect tooth formation, and give us that beautiful smile. Formation of inorganic crystal structures by living organism in tooth bio mineralization is a substantial scientific puzzle. The ability of cells to form highly defined polymorphic crystal structures, while the living cells need precise molecular control and efficient mechanisms is nothing but magnificent. Amg has not only a critical role in correct enamel formation, but also has multifunctional characteristics of wound healing, anti-inflammatory, anti-infectious and signaling properties in living organisms, which make it a vital protein in biomedical research. Moreover, new advances in Amg production methodology have made it a promising tool for developing new materials, thus enriching this field of research. Amg has a natural characteristic of pH dependent self-assembly into several nanospheric structures. Nanoparticles made from recombinant proteins, can have many important applications in biotechnology, but most of them actually could not be used in industry due to low stability and/or low availability. On the other hand, the simple production of self-assembled Amg nanospheres can overcome these drawbacks. Amg nanospheres can be constructed from pure Amg or from an Amg fusion protein, with a diameter of ~30-40 nm (for recombinant Amg). The fused domain can be for example cellulose binding domain or another protein or peptide with a significant application or function that can be added to the system. The current study provides the evidence of Amelogenin nanoparticles high affinity for binding to cellulosic materials under simply controlled conditions. This study has suggested potential use in drug delivery systems applications and in textiles surface modification in industry. Consequently, more applications can be explored and more advances can be achieved. By labeling Amg with fluorescent dye, binding can also be examined and monitored using the fluorescence microscopy.