Structural characterization of Kannurin isoforms and evaluation of the role of β-hydroxy fatty acid tail length in functional specificity

The novel anti-fungal cyclic lipopeptide ‘Kannurin’ and its three structural variants produced by Bacillus cereus AK1 were previously reported from our laboratory. The present study reports unexplored structural variants of Kannurin those have functional benefits. Due to the difference in β-hydroxy fatty acid tail length, they are designated here as Kannurin A (m/z 994.67 ± 0.015), B (m/z 1008.68 ± 0.017), C (m/z 1022.69 ± 0.021), D (m/z 1036.70 ± 0.01), C(L) (m/z 1040.71 ± 0.02) and D(L) (m/z 1054.72 ± 0.01). The isoform A (m/z 994.67 ± 0.015) is the shortest cyclic form of Kannurin identified so far. In addition, C(L) (m/z 1040.71 ± 0.02) and D(L) (m/z 1054.72 ± 0.01) are the rare natural linear forms. The results of the antimicrobial assays deduced that the difference in lipid tail length of the isoforms contributes tremendous differences in their antimicrobial properties. The isoforms with short lipid tails (A and B) are more selective and potent towards bacteria, whereas the isoforms with long lipid tails (C and D) are more potent against fungi. The molecular dynamics studies and electron microscopic observations supported with circular dichroic spectroscopy analysis showed the structural confirmation and formation of aggregates of Kannurin in solution. The molecular dynamics simulation studies revealed that a single molecule of Kannurin makes enormous intra-molecular interactions and structural re-arrangements to attain stable lowest energy state in solution. When they reach a particular concentration (CMC) especially in aqueous environment, tends to form structural aggregates called ‘micelles’. With the structural information and activity relationship described in this study, it is trying to point out the sensitive structural entities that can be modified to improve the efficacy and target specificities of lipopeptide class of antibiotics.