Molecular and culture-based characterization of bacterial community from Manleluag Alkaline Spring in Pangasinan [Philippines]: project title: isolation and screening of alkalne bacteria from Manleluag Alkaline Spring producing and industrially important enzyme

A total of 826 bacterial isolates were cultured and characterized from three sample collection locations in Manleluag hyperalkaline spring in Mangatarem, Pangasinan [Philippines]. Sampling was also done in 4 different quarters to obtain a thorough presentation of the bacterial population of the area. Variation in the cultural morphology of the isolates were observed, with majority displaying Gram-positive reaction (90%) typical in environments with elevated pH conditions. Bacterial isolates were screened for the presence of different enzymes of industrial importance. A total of 428 bacterial isolates showed protease activity when tested using Skim Milk Assay. Majority of these isolates was obtained during the Q2 sampling Protease activity was evaluated at two pH conditions (pH 7 and pH 10). Isolate SNE-IV-1.2-137 had the highest protease activity (Cz ratio = 0.2056). Cyclodextrin glycosyltransferase (CGTase) activity was evaluated using Phenolphthalein Assay, with 54 bacterial isolates exhibiting significant activity. Isolate WNE-I-1.1-036 displayed highest CGTase activity (46.0094 U/ml). Asparaginase activity of the bacterial isolates were initially screened using qualitative assay using colorimetric method. Ninety seven isolates initially screened using a qualitative assay using colorimetric method. Ninety seven isolates initially displayed enzyme activity which was consequently verified using quantitative Nesslerization method. Isolate WNE-II-1.3-121 had the highest enzyme activity of 26 mM NH sub 3 per ml enzyme sample at 24 hours. Presence of antimicrobial activities were done using 16S rDNA sequencing. Lastly, preliminary optimization of protease extraction was also done using isolate SNE-IV-1.2-137. Protease extraction and was observed at its highest using 70% ammonium sulfate saturation.