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.

Wound healing by topical application of Momordica charantia L. formulations on mice

“…M. charantia contains compounds such as momorcharin, momordenol, momordicillin, momordicinin I, II and III, momordine, momordolol, charantin, charine, cryptoxanthin, cucurbitans, cycloartenol, diosgenin, eleostearic acid, erythrodiol, galacturonic acids, gentisic acid, goyasaponin, and multiflorenol [ 15 - 18 ]; which are responsible for its various biological and pharmacological effects such as antidiabetic [ 19 ], antiulcer [ 20 ], antimicrobial [ 21 ], and antioxidant [ 22 ]. In addition, MAP30 protein that shows anti-HIV activity has also been identified in M. charantia [ 23 ].…”

Anti-viral chimeric protein RetroMAD1™ potently block SARS-CoV-2 viral entry and propagation

“…Extracted from Momordica charantia seeds, MAP-30 is reported to possess anti-HIV infection and replication [17,18], and anti-tumor activity (Lee-Huang et al, 1995, Lee-Huang et al, 2000, Jiang et al, 2018 inhibiting the infection and replication of Herpes Simplex Virus HSV-1, HSV-2 as well as acyclovir-resistant HSV strains [21]. MAP-30 also signi cantly reduced the dosage of chloramphenicol and erythromycin required to inhibit the growth of Staphylococcus aureus, Enterococcus faecalis, Salmonella typhimurium, Salmonella enteritidis, and Pseudomonas aeruginosa [22], inhibits the growth of Kaposi's sarcoma-associated herpes virus (KSHV)-infected tumor cells from AIDS patients by inhibiting the expression of viral and cellular genes essential for proliferation and expansion of the virus infected tumor cells [23], and effectively suppressing Hepatitis B virus (HBV) gene expression and genome replication [24]. Belonging to the family of type-1-ribosomal inactivating proteins (RIPs), MAP-30 has N-glycosidase activity that depurinates adenine base-ribose glycosidic bond at position A-4324 in universally conserved α-sarcin /ricin loop of 28S ribosomal (r)RNA [25,26].…”

High-yield expression in Escherichia coli, biophysical characterization, and biological evaluation of plant toxin gelonin

“…Given the ineffectiveness of small-molecule chemotherapeutic agents, macromolecular drugs have attracted growing interest over the past few decades because of their unparalleled potency and unique mode of action (Ye et al 2015). One representative example is the plant-derived ribosome-inactivating proteins (RIPs), which exhibits a glycosidase activity and can be classified into three groups referred to as types I, II, and III (Chang et al 2017). Type I RIPs consist of a single polypeptide chain (A-chain) with catalytic activity, type II RIPs are heterodimeric glycoproteins composed of an A-chain (similar to type I RIPs) linked to a B chain (mediating cellular internalization) via disulfide bond.…”

“…Type I RIPs consist of a single polypeptide chain (A-chain) with catalytic activity, type II RIPs are heterodimeric glycoproteins composed of an A-chain (similar to type I RIPs) linked to a B chain (mediating cellular internalization) via disulfide bond. Type III RIPs comprise an A-chain and an additional protein fragment that requires a proteolytic cleavage to give active RIPs (Stirpe 2013;Chang et al 2017). RIPs hold great potential for biomedical applications due to their various biological activities such as antiviral, antiparasitic, and anticancer properties (Stirpe 2006).…”