Evaluation of L-Asparaginase Purified from Pseudomonas Fluorescens as Inhibitor for Biofilm Producers in Dental Decays.

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This study aimed to assess Pseudomonas fluorescens-purified L-Asparaginase's effectiveness as a broad-spectrum inhibitor of biofilm producers in dental decays. The 16S rRNA sequence was used to build a phylogenetic tree to calculate the evolutionary distance between the isolated bacterial strain SW3 and other species. The evolutionary history was inferred by using the neighbor-joining approach. The bacteria were identified from dental decays, including Staphylococcus aureus, Streptococcus mutans, Streptococcus oralis, and Streptococcus mitis. Each one of these isolates showed different degrees of biofilm development. Purified L-Asparaginase inhibited the most potent Gram-positive biofilm-forming bacteria (biofilm producers) with higher inhibition percentages against Streptococcus oralis and Streptococcus mitis, 65 - 73.8 % and 54.7 - 63%, respectively. The inhibition percentages increased with increasing concentration and reached up to 74 - 81% with Streptococcus oralis and 66 - 74% with Streptococcus mitis, while SW3 bacteria showed (100%). This strain was suggested SW3 (Pseudomonas spp.). Pseudomonas fluorescens bacterial strain isolated from rhizosphere soil produced extracellular L-Asparaginase when grown on as a substrate. L-Asparaginase was purified to homogeneity by using ammonium sulfate at 60% saturation, followed by gel filtration chromatography on a sephadex G-100 column, with a recovery yield of 49% and a purification fold of 2.22. L-Asparaginase had a promising use for removing and avoiding biofilm growth, implying that it might be used in the dental industry in the future.