In this study, screening and isolation of potent L- glutaminase producing halotolerant bacteria was done. This strain was identified as Bacillus halotolerans PHN1. It was observed that organisms have the ability to synthesize and produce L-glutaminase enzyme at 3% NaCl concentration and can withstand with 7% of NaCl concentration. Maximum enzyme production was at 96 h of its growth. Partial purification of this enzyme was carried out at 60 % ammonium sulphate precipitation followed by dialysis and then Ion exchange chromatography. The overall yield obtained after purification is 4.68% with 1.56 U/mg of specific activity. The molecular weight of the enzyme was shown 60 kDa confirmed by SDS-PAGE electrophoresis. The enzyme has optimum activity at pH 7.0 with 30ºC. It is strongly inhibited by various metal ions included Hg+2; Ag+2, Zn+2, (percentage inhibition obtained was 60-70% with a concentration of metal ion taken was 25µg, 50µg and 75µg/ ml respectively) Mg2+ and Mn2+ concentration dependent manner. Km and Vmax for enzyme are 0.014 mM and 0.07U/ml respectively.