• Microbiology and Biotechnology Letters
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• v.27 no.6
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• pp.440-445
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• 1999

In order to investigate structure-antibiotic activity relationships of brevinin-1 and thanatin containing Rana box composed of basic loop formed by disulfide bridge in their arboxy terminus, thanatin, brevinin 1 and their analogues (T-B1, T-B2 and B-T) in which their Rana box sequence exchanged was designed and synthesized by the solid phase method using Fmoc-chemistry. The basic sequence of Rana box of thanatin had more significant effect on both antibacterial and antifungal activity than that of brevinin 1. The tail sequence (QRM) of thanatin was found to be important in its antibacterial and antifungal activity. Rana box sequence of brevinin-1 did not have a significant effect on its antitumor and phospholipid vesicle-aggregating activities. Brevinin-1 showed stronger $\alpha$-helical structure in the membrane-mimicking environment such as SDS micelle than thanatin. A remarkable increase in a-helicity of bervinin-1 plays more important role in antibiotic activity than that of thanatin. Furthermore, antibacterial activity of thanatin against E. coli resulted from the disruptive effect against the outer cell membrane of E. coli.

• BMB Reports
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• v.35 no.3
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• pp.291-296
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• 2002

Thanatin, a 21-residue peptide, is an inducible insect peptide with a broad range of activity against bacteria and fungi. It has a C-terminal disulfide loop, like the frog skin secretion antimicrobial peptides of the brevinin family. In this study, we tried to find the effect of a number of amino acids between the disulfide bond. Thanatin showed stronger antibacterial activity to Gram negative bacteria than other mutants, except Th1; whereas, the mutant peptides with deletion had higher activity to Gram positive bacteria than thanatin. An increase in the number of amino acid(s) using the alanine residue decreased the antibacterial activity in all of the bacteria. Th1 with deletion of threonine at position 15 ($Thr^{15}$) showed similar antibacterial activity against Gram-negative bacteria, but had higher activity against the Gram positive bacteria. In order to study the structure-function relationship, we measured liposome disruption by the peptides and CD spectra of the peptides. Th1 also showed the highest liposome leaking activity and α-helical propensity in the sodium dodecyl sulfate solution, compared with other peptides. Liposome disruption activity was closely correlated with the anti-Gram positive bacterial activity. All of the peptides showed no hemolytic activity. Th1 was considered to be useful as an antimicrobial peptide with broad spectrum without toxicity.