title: Novel purification method and antibiotic activity of recombinant Momordica charantia MAP30.
abstract: Ribosome-inactivating proteins (RIPs) are a group of enzymes originally isolated from plants that possess the ability to damage ribosomes in an irreversible manner, leading to inhibition of protein synthesis in eukaryotic cells. In this study, we aimed to purify recombinant RIPs, investigate their function in the treatment of bacterial infection, and determine their toxicity in mice. We employed a pMAL protein fusion and purification system using E. coli transformed with a plasmid containing MBP-tagged MAP30 cDNA. MBP-tagged MAP30 was purified using a modified novel protocol to effectively produce highly active MAP30 of high purity. In an acute toxicity study in mice, no mortality occurred at doses lower than 1.25 mg/kg. MAP30 at both 0.42 and 0.14 mg/kg induced anti-MAP30 IgG, which reached a maximum titer at week 3. In conclusion, recombinant MAP30 prepared using our purification method possesses bioactivity, and has a synergistic bacteria-killing effect that can significantly reduce the required dosages of chloramphenicol and erythromycin. Therefore, when MAP30 is used in combination with chloramphenicol or erythromycin, it may of benefit in terms of reducing the side effects of the antibiotics, as lower concentrations of antibiotics are required.
authors.affiliation: Department of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung, Taiwan.
authors.forename: Ching-Dong
authors.initials: CD
authors.lastname: Chang
authors.affiliation: Department of Biological Science and Technology, National Pingtung University of Science and Technology, 1, Shuefu Rd., Neipu, Pingtung, 91201, Taiwan.
authors.forename: Ping-Yuan
authors.initials: PY
authors.lastname: Lin
authors.affiliation: Department of Biological Science and Technology, National Pingtung University of Science and Technology, 1, Shuefu Rd., Neipu, Pingtung, 91201, Taiwan.
authors.forename: Yo-Chia
authors.initials: YC
authors.lastname: Chen
authors.affiliation: Department of Veterinary Medicine, National Chiayi University, Chiayi City, Taiwan.
authors.forename: Han-Hsiang
authors.initials: HH
authors.lastname: Huang
authors.affiliation: Department of Biological Science and Technology, National Pingtung University of Science and Technology, 1, Shuefu Rd., Neipu, Pingtung, 91201, Taiwan. wlshih@mail.npust.edu.
authors.forename: Wen-Ling
authors.initials: WL
authors.lastname: Shih
keywords: Antibiotic activity; MAP30; Mormodica charantia; Ribosome-inactivating proteins