A halo-tolerant glutaminase gene (BlglsA) was isolated from Bacillus licheniformis. Heterologous expression of BlglsA revealed that it encodes for a 36 kDa protein containing 327 amino acid residues. The purified enzyme showed optimal activity at a pH of 9.5 while 35 °C was found to be the optimum temperature. The enzyme retained about 92 and 97% stability at pH 12 and temperature (40 °C) respectively. Subsequent immobilization of BlglsA on nano magnetic cellulose sheet (NMCS) led to an enhanced tolerance to higher temperature. NMCS-BlglsA showed optimum activity at 45 °C, although it was stable even at 60 °C. NaCl tolerance (≥90% in 0.3 M) was almost similar to BlglsA and NMCS-BlglsA. The metal ions Fe2+ (5 mM) and Mn2+ (2.5 mM) improved the BlglsA relative activity by 61 and 48%, respectively. In contrast, 5 mM Mn2+ was found suitable to enhance the activity of NMCS-BlglsA up to 72%. The production of glutamic acid by NMCS-BlglsA was 1.61 g/l in 48 h. Reusability test of NMCS-BlglsA showed 76 and 35% retention of the actual activity after 4th and 7th cycle, respectively. Such remarkable biochemical properties of NMCS-BlglsA make it an attractive enzyme for food industries.
Keywords: Bacillus licheniformis; Glutamic acid; Glutaminase; Immobilization; Thermostability.
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