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Halophiles produce stable enzymes under extreme conditions. The scant information about chitinolytic haloarchaea led us to conduct the present study in order to isolate and screen native halophilic archaea with chitinolytic activity and to optimize the enzyme production conditions. Among 500 haloarchaeal strains isolated from water samples from different hypersaline lakes of Iran, five strains showed chitinolytic activity. Based on biochemical, morphological and molecular analyses, we established that all five potent strains belonged to the genus Natrinema. Besides, observing chitinase function in culture media, through an additional molecular test the presence of the chitinase gene in chitinase-producing strains was also confirmed by PCR amplification. Compared with other potent strains, Natrinema sp. strain BS5 showed significant chitinase production. The production of chitinase in strain BS5 accompanied growth, started at the logarithmic phase and increased to its maximum level at the beginning of the stationary phase. Maximum chitinase production was obtained at 37˚C, pH 7.5, 3 M NaCl and 1% colloidal chitin. The strain BS5 showed 38%, 30%, 24% and 28% decreases in enzyme production at 40˚C, pH 8, 3.5 M NaCl and 0.5% substrate, respectively. This strain was able to produce the enzyme in NaCl 4 M and in the absence of MgCl 2 and MgSO 4. This study revealed the strong potential of the genus Natrinema to produce chitinase at high salt concentrations without Mg 2+ requirement.

Groundwater pollution is one of the major environmental concerns. The entrance of pollutants into the oligotrophic groundwater ecosystems alters native microbial community structure and metabolism. This study investigated the application of innovative Small Bioreactor Chambers and CaO2 nanoparticles for phenol removal within continuous-flow sand-packed columns for 6 months. Scanning electron microscopy and confocal laser scanning microscopy analysis were conducted to indicate the impact of attached biofilm on sand surfaces in bioremediation columns. Then, the influence of each method on the microbial biodiversity of the column’s groundwater was investigated by next-generation sequencing of the 16S rRNA gene. The results indicated that the simultaneous application of biostimulation and bioaugmentation completely eliminated phenol during the first 42 days. However, 80.2% of phenol remained in the natural bioremediation column at the end of the experiment. Microbial diversity was decreased by CaO2 injection while order-level groups known for phenol degradation such as Rhodobacterales and Xanthomonadales dominated in biostimulation columns. Genome-resolved comparative analyses of oligotrophic groundwater prokaryotic communities revealed that Burkholderiales, Micrococcales, and Cytophagales were the dominant members of the pristine groundwater. Six-month exposure of groundwater to phenol shifted the microbial population towards increasing the heterotrophic members of Desulfobacterales, Pseudomonadales, and Xanthomonadales with the degradation potential of phenol and other hydrocarbons.