Among several bacteria examined in this study, a hyper acidophil and thermostable Micrococcus sp.NS 211 designated as M.Amy NS 211 was selected for production of amylase using starch agar plates with following incubation at 85°C. Identification by 16SrRNA on selected bacterium disclosed the highest similarity for protean regions of this gene, 27 F and 1492R as Micrococcus sp.NS 211. Although activity of M.Amy NS 211 was established at temperatures between 70 and 110°C and pH ranges 1.2–8.0, the optimum temperature and pH was achieved at 85°C and 3.5 in sodium citrate buffer system respectively. Two‐step chromatography was performed using (CM Bio‐Gel A) and (Bio‐Gel A‐150) columns to purify 84 kDa hyper acidophil and thermostable α‐amylase. SDS‐PAGE analysis showed molecular mass and amylolytic activity as single band. Enhancement of enzyme activity was obtained in presence of 5 mM MnCl2 (298%), CaCl2 (347%), FeCl2 (211%), MgCl2 (253%), ZnCl2 (146%), NiCl (142%), NaCl (141%), Na‐sulfate (153%) while inhibition was observed with (5 mM) EDTA, PMSF (3 mM), urea (8 M), and SDS (1%) at 143, 134, 43, and 119%, respectively. M.Amy NS 211 can be applied in laundry detergents, textile, and modern relevant industrial processes at extreme temperatures and under acidic conditions.

Our newly discovered metalloprotease, designated as ALP NS12 was selected using gelatin agar plates with incubation at 100 °C. Subcloning of the fragments in to pUC118 to make E. coli HB101 (pPEMP01NS) with following two-step chromatography using diethylaminoethyl sepharose (DEAE-sepharose) and Sephadex G-100 columns to purify 97-kDa expressed enzyme was performed. Although activity of immobilized ALP NS12 on glass surface was established at temperatures between 70 and 120 °C and pH ranges 4.0-13.0, the optimum temperature and pH were achieved at 100 °C and 11.0, respectively. Enhancement of enzyme activity was obtained in the presence of 5 mM MnCl2 (91 %), CaCl2 (357 %), FeCl2 (175 %), MgCl2 (94 %), ZnCl2 (412 %), NiCl (86 %), NaCl (239 %), and Na-sulfate (81 %) while inhibition was observed with EDTA (5 mM), PMSF (3 mM), urea (8 M), and SDS (1 %) at 65, 37, 33, and 42 %, respectively. Consequently, the enzyme was well analyzed using crystallography and protein modeling. ALP NS12 can be applied in industrial processes at extreme temperatures and under highly basic conditions, chelators, and detergents.