title: Expression of a Recombinant Anti-HIV and Anti-Tumor Protein, MAP30, in Nicotiana tobacum Hairy Roots: A pH-Stable and Thermophilic Antimicrobial Protein.
abstract: In contrast to conventional antibiotics, which microorganisms can readily evade, it is nearly impossible for a microbial strain that is sensitive to antimicrobial proteins to convert to a resistant strain. Therefore, antimicrobial proteins and peptides that are promising alternative candidates for the control of bacterial infections are under investigation. The MAP30 protein of Momordica charantia is a valuable type I ribosome-inactivating protein (RIP) with anti-HIV and anti-tumor activities. Whereas the antimicrobial activity of some type I RIPs has been confirmed, less attention has been paid to the antimicrobial activity of MAP30 produced in a stable, easily handled, and extremely cost-effective protein-expression system. rMAP30-KDEL was expressed in Nicotiana tobacum hairy roots, and its effect on different microorganisms was investigated. Analysis of the extracted total proteins of transgenic hairy roots showed that rMAP30-KDEL was expressed effectively and that this protein exhibited significant antibacterial activity in a dose-dependent manner. rMAP30-KDEL also possessed thermal and pH stability. Bioinformatic analysis of MAP30 and other RIPs regarding their conserved motifs, amino-acid contents, charge, aliphatic index, GRAVY value, and secondary structures demonstrated that these factors accounted for their thermophilicity. Therefore, RIPs such as MAP30 and its derived peptides might have promising applications as food preservatives, and their analysis might provide useful insights into designing clinically applicable antibiotic agents.
authors.affiliation: Institute of Biotechnology, Shiraz University, Shiraz, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Moghadam
authors.affiliation: Institute of Biotechnology, Shiraz University, Shiraz, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Niazi
authors.affiliation: Plant Virology Research Center, College of Agriculture, Shiraz University, Shiraz, Iran.
authors.forename: Alireza
authors.initials: A
authors.lastname: Afsharifar
authors.affiliation: Department of Plant Protection, College of Agriculture, Shiraz University, Shiraz, Iran.
authors.forename: Seyed Mohsen
authors.initials: SM
authors.lastname: Taghavi
mesh_terms: D000595:Amino Acid Sequence; D019380:Anti-HIV Agents / Q000494:pharmacology; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology; D017124:Conserved Sequence; D004926:Escherichia coli / Q000187:drug effects; D015870:Gene Expression; D023061:Gene Order; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D008826:Microbial Sensitivity Tests; D018517:Plant Roots / Q000235:genetics*; D030821:Plants, Genetically Modified; D010957:Plasmids / Q000235:genetics; D056510:Position-Specific Scoring Matrices; D054730:Protein Interaction Domains and Motifs; D055550:Protein Stability; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D054790:Ribosome Inactivating Proteins, Type 2 / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D013696:Temperature; D014026:Nicotiana / Q000235:genetics*
chemical_list: D019380:Anti-HIV Agents; D000972:Antineoplastic Agents, Phytogenic; C066347:MAP30 protein, Momordica charantia; D011994:Recombinant Proteins; D054790:Ribosome Inactivating Proteins, Type 2