Molecular Characterization, Structural Modeling, and Evaluation of Antimicrobial Activity of Basrai Thaumatin-Like Protein against Fungal Infection

• Published in: BioMed research international 2017-01, Vol.2017 (2017), p.1-9
• Main Authors: Gul, Roquyya, Naz, Mamoona, Saleem, Mahjabeen, Yasmin, Nusrat, Rehman, Hafiz Muzzammel
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Amphotericin B; Antifreeze proteins; Antifungal activity; Antifungal agents; Antifungal Agents - chemistry; Antifungal Agents - therapeutic use; Antiinfectives and antibacterials; Antimicrobial activity; Antimicrobial agents; Banana; Cloning; Crystallization; Deoxyribonucleic acid; DNA; Docking; E coli; Ergosterol; Escherichia coli; Frozen food; Frozen foods; Fungi; Fungi - drug effects; Fungi - pathogenicity; Fungicides; Gene expression; Gene Expression Regulation, Plant; Genetic aspects; Health aspects; Humans; Ice; Ice formation; Infections; Membrane proteins; Molecular biology; Molecular Docking Simulation; Molecular weight; Musa - chemistry; Musa - genetics; Mycoses; Mycoses - drug therapy; Mycoses - microbiology; Pathogenesis; Pharmacology; Plant proteins; Plant Proteins - chemistry; Plant Proteins - genetics; Plant Proteins - therapeutic use; Plasmids; Preservation; Prevention; Protein Conformation; Proteins; Sequence Homology, Amino Acid; Structural analysis; Thaumatin; Yogurt
• ISSN: 2314-6133; 2314-6141
• DOI: 10.1155/2017/5046451

A thaumatin-like protein gene from Basrai banana was cloned and expressed in Escherichia coli. Amplified gene product was cloned into pTZ57R/T vector and subcloned into expression vector pET22b(+) and resulting pET22b-basrai TLP construct was introduced into E. coli BL21. Maximum protein expression was obtained at 0.7 mM IPTG concentration after 6 hours at 37°C. Western blot analysis showed the presence of approximately 20 kDa protein in induced cells. Basrai antifungal TLP was tried as pharmacological agent against fungal disease. Independently Basrai antifungal protein and amphotericin B exhibited their antifungal activity against A. fumigatus; however combined effect of both agents maximized activity against the pathogen. Docking studies were performed to evaluate the antimicrobial potential of TLP against A. fumigatus by probing binding pattern of antifungal protein with plasma membrane ergosterol of targeted fungal strain. Ice crystallization primarily damages frozen food items; however addition of antifreeze proteins limits the growth of ice crystal in frozen foods. The potential of Basrai TLP protein, as an antifreezing agent, in controlling the ice crystal formation in frozen yogurt was also studied. The scope of this study ranges from cost effective production of pharmaceutics to antifreezing and food preserving agent as well as other real life applications.