Features of bone destruction due to the collagenase activity of bacteria of the Bacillus and Clostridium genera

Abstract

The purpose of work was to study the activity of collagenase biosynthesis by bacilli and clostridia isolated from the bone microflora in a pure culture at different periods of bone decomposition and at different values of acidity of the medium and the temperature of cultivation. The comparative collagenase activity of the strains of Bacillus mycoides, Bacillus subtilis, Clostridium putrificum, Clostridium sporogenes found in microflora of bone tissue was determined. The catalytic activity of collagenases was evaluated according to the modified method of agar blocks by the diameter of precipitation zone as a result of diffusion of enzyme into an agar medium with collagen. Within 6 months of experiment, the dynamics of changes in the number of collagenolytic strains was monitored. For all isolated Bacillus and Clostridium isolates, a steady increase in collagenase synthesis was observed throughout the study period, with the exception of Bac. subtilis, which is probably due to the acidity of medium, which did not correspond to the optimal pH range for this species. The optimum temperature for manifestation of the maximum collagenolytic activity of bacilli was 40 °C, and for clostridia - 30 °C. The study of proteolytically active species involved in the destruction of bone collagen is promising for the purposes of forensic examination. The use of bacterial enzymes in forensic practice remains a difficult task, however, the use of their substrate specificity can significantly expand the evidence base of the examinations.