Purification of Bacteriocins by Chromatographic Methods

Автор: V.I. Surovtsev, V.M. Borzenkov, and V.P. Levchuk

Страница: 44-49

Purification of Bacteriocins by Chromatographic Methods

Russian Journal of Biotechnology, 2014, N 5, P. 44-49

UDC 577.21:577.352:579.891.1

Section:  “Biologicals Technology”

V.I. Surovtsev,  V.M. Borzenkov ^*,  and  V.P. Levchuk

The State Research Center for Applied Microbiology and Biotechnology,  142269, Obolensk, Moskovskaya Oblast Russia

e-mail:  vmborzenkov@mail.ru

The bacteriocins secreted by cells of the two B.subtilis strains have been isolated by hydrophobic or ion-exchange chromatography on CMC followed by the purification on membranes. The yields of electrophoretically pure products were ~ 75 % of the total activity in the supernatant of culture liquid that was obtained after the cell sedimentation. A bacteriocin secreted by B. subtilis BSX strain cells has high hydrophobicity and it is mainly found out on the surface of cell walls in the culture liquid. These fragments served as sorbents in hydrophobic chromatography. A B. subtilis strain B-112 bacteriocin predominantly occurs in the free state in the supernatant of culture liquid. It was purified after the removal of the cell debris on CMC followed by the elution with a glycine-HCl buffer, pH 2,5. The concentration of the purified bacteriocins was determined at 205 nm and therefore the method did not require the availability of aromatic amino acids in the macromolecule structure. It is believed that these chromatographic techniques can be used for the purification with a high yield of bacteriocins of other producers.

Key words:  bacteriocins,  hydrophobicity,  ion-change and hydrophobic chromatography,  membranes,  purification,  surface cell fragments.

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