I have sigma nisin 2.5% (balance sodium chloride and denatured milk solids) according to specification it dissolves in water, but I'm getting some precipitation while dissolving in water or 5% DMSO. Please give some suggestion to dissolve it properly. I have to use it for cell culture purposes.
Thanks
Nisin is active against Gram-positive bacteria and exerts two killing mechanisms. Firstly, nisin inhibits cell-wall synthesis by binding to lipid II, an essential membrane-anchored cell-wall precursor, and secondly, nisin permeabilizes the target membrane. The binding of nisin to lipid II induces membrane integration of nisin resulting in the formation of a pore, likely composed of eight nisin and four lipid II molecule.Nisin from L. lactis, like almost all naturally produced AMPs, can be purified directly from the culture medium.It is a cationic peptide, and therefore, commonly purified using cation exchange chromatography (cIEX) at acidic pH, using high salt concentration for elution, typically a single-step elution with 1 M NaCl .The column is subsequently washed with 50 mM lactic acid pH 3 until a stable baseline is reached to remove nonspecifically bound material. Peptides are eluted by increasing the NaCl concentration stepwise using a flow rate of 1 mL/min, resulting in elution fractions with 200 mM (Step I), 400 mM (Step II), 600 mM (Step III), 800 mM (Step IV) and 1 M (Step V) NaCl. To remove NaCl, protein in the elution fractions is precipitated with 20% (v/v) trichloroacetic acid (TCA) overnight at 4°C. Precipitated protein is washed two times with ice-cold acetone to remove residual TCA. Finally, the protein pellet is suspended in 50 mM lactic acid pH 3. Nisin concentrations can be determined by a colorimetric assay (Pierce BCA Protein Assay Kit, Thermo scientific) by measuring the absorbance at 584 nm according to the protocol of the manufacturer.
you may add some organic acids depending on your work, for example th procedure you want to assay it.
You could also use 0.05% acetic acid. I make up my nisin with 0.05% acetic acid for induction purposes.
Hi Dr. Anil, I have purchased the same product from sigma, Nisin 2.5%, how could you dissolve it? would you please tell me ... Thank you.
Dissolve 40 mg of 2.5% nisin in 1 mL of 0.05% acetic acid, leave for sometime. The milk proteins will not dissolve (dirty solution), centrifuge at 10,000 rpm for 1 min on mini tabletop centrifuge (turns clear) and transfer the supernatant to a new MCT, filter sterilize and aliqout at -20.
Hi Anıl,
Could you give your suggestions to dissolve nisin (sigma) properly if you found out?
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