Optimization of the culture conditions for the production of a bacteriocin from halophilic archaeon Sech7a.

An extremely halophilic haloarchaeon Sech7a, isolated from a solar saltern, was found to excrete halocin, a bacteriocin like substance. Optimal antimicrobial activity was obtained at 45 degrees C using 0.5% (w/v) glycerol and 0.5% (w/v) yeast extract as nutrients in SW media containing 3.4 M NaCl with pH value 7.5. Halocin Sech7a is a 10.7-kDa polypeptide, which is stable in a wide range of pH and is thermolabile at temperatures above 80 degrees C. As many other halophilic proteins, halocin Sech7a loses part of its activity upon exposure to low salt conditions, yet its activity can be restored after dialysis against initial saline conditions. Microscopic inspection revealed swelling and lysis of sensitive cells upon exposure to halocin Sech7a. These results indicate that haloarchaeon Sech7a excretes a novel bacteriocin.

Characterization of parazoanthoxanthin A binding to a series of natural and synthetic host DNA duplexes.

Parazoanthoxanthin A is a fluorescent yellow nitrogenous pigment of the group of zoanthoxanthins, which show a broad range of biological activity. These include, among others, the ability to bind to DNA. In this study we have used a variety of spectroscopic (intrinsic fluorescence emission and UV-spectroscopy) and hydrodynamic techniques (viscometry) to characterize in more detail the binding of parazoanthoxanthin A to a variety of natural and synthetic DNA duplexes in different buffer conditions. Our results reveal the following five significant features: (i) Parazoanthoxanthin A exhibits two modes of DNA binding: One binding mode exhibits properties of intercalation, while the second binding mode is predominantly electrostatic in origin. (ii) The apparent binding "site size" for parazoanthoxanthin A near physiological salt concentration (100 mM NaCl) is in the range of 7 +/- 1 base pairs for natural genomic DNA duplexes (calf thymus and salmon testes DNA) and alternating synthetic polynucleotides (poly[d(AT)]. poly[d(AT)] and poly[d(GC)]. poly[d(GC)]). A slightly larger apparent binding site size of 9 +/- 1 bp was obtained for parazoanthoxanthin A binding to the synthetic homopolymer poly[d(A)]. poly[d(T)]. (iii) Near physiological salt concentration (100 mM NaCl) parazoanthoxanthin A binds with the same approximate binding affinity of 2-5 x 10(5) M(-1) to all DNA polymers studied. (iv) At low salt concentration, parazoanthoxanthin A preferentially binds alternating poly[d(AT)]. poly[d(AT)] and poly[d(GC)]. poly[d(GC)] host duplexes. (v) Parazoanthoxanthin A inhibits DNA polymerase in vitro.