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After 48 h, biofilms were washed once with 1 PBS followed by treatment with different enzyme solutions prepared in 1 PBS, pH 7.4, including: 5, 10, 20, and 40U DNase-I, 100, 200, 500 and 1000 g/ml amylase and 50, 100, 200 and 500 g/ml Proteinase K, for 24 h, at 37C and 150 rpm, the control was treated with 1 PBS alone

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Further structural optimization delivered a large number of macrocyclic disulfides and among them 23 (HA08) encompassing a 3-homotyrosine residue in a 13-membered ring system and exhibiting the lowest Ki value in the series (Ki = 3.3 nM)