Purification and functional characterization of recombinant balsamin, a ribosome-inactivating protein from Momordica balsamina

Parminder Ajji, Shailendra Sonkar, Ken Walder, Munish Puri

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

Balsamin, a type I ribosome-inactivating protein (RIP), has been shown to inhibit HIV-1 replication at the translation step. Our recent studies have shown that balsamin also possess anti-tumor, antibacterial and DNase-like activity, however, the amount of natural balsamin in Momordica balsamina seeds is limited and preclinical studies require large quantities of pure, bioactive balsamin. Therefore, in this study, we cloned the balsamin gene, expressed it in E. coli BL21 (DE3) strain and purified it by nickel affinity chromatography. Functional analysis indicated that balsamin exhibits both RNA N-glycosidase activity, releasing the Endo-fragment from rabbit reticulocyte rRNA, and DNase-like activity, converting the supercoiled form of a plasmid into the linear form in a concentration-dependent manner. Analysis of secondary structure revealed that recombinant balsamin mainly consisted of α-helical and random coiled with minimal turns and β-sheets. Recombinant balsamin was found to be stable in the temperature range of 20–60 °C and pH range of 6–9. Antimicrobial assays showed that the minimum inhibitory concentrations of recombinant balsamin for various pathogens ranged between 1.56 and 12.5 μg/ml. Heterologous expression and purification of balsamin carries great importance as it provides an alternative approach for large-scale preparation of biologically active recombinant balsamin, which is difficult from its natural source.

Original languageEnglish
Pages (from-to)2 26-234
Number of pages9
JournalINTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
Volume114
Issue number2018
DOIs
Publication statusPublished - 15 Jul 2018

Keywords

  • Antimicrobial and heterologous expression
  • Bioactive
  • DNase
  • Nutraceutical
  • RNA N-glycosidase

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