Ribonuclease a

BAC-IB17 was also treated with various enzymes at 37 °C and 50 °C up to 24 h including protease (MP Biomedicals, USA), proteinase-K (Life Technologies, USA), amylase (Sigma-Aldrich, USA), pepsin, lysozyme, ribonuclease A and catalase (Serva, Germany). For this 1.0 mL of purified bacteriocin with initial concentration of 0.43 mg ml^− 1 was incubated with equal volume of enzymes with the final concentration of all the enzymes used was 1.0 mg ml^− 1 and the relative antibacterial activity was calculated using agar well diffusion assay.

CHO and DLD-1 cell lines
were purchased from
ATCC. Dulbecco Modified Medium (DMEM; Biological Industries), Fetal
Bovin Serum (FBS; Biological Industries), l-glutamine, and
Penicillin/Streptomycin were used as complete cell culture mediums.
Cells were removed from the culture dishes and washed with trypsin-EDTA
(trypsin-ethylenediamine tetraacetic acid) and phosphate buffer (PBS).
Cells were counted using trypan blue. In a cytotoxicity test, cell
viability was determined by using MTT, a tetrazolium salt. In dual
staining, apoptosis/necrosis ratios were determined using Ribonuclease-A,
Propidium Iodide (Serva, Israel), and Hoechst 33342 (Serva, Israel).
All cell culture studies were performed in culture dishes and multiwell
plates (Corning, USA).