They can be cultivated under extreme pH conditions and these species produce extracellular enzymes that are resistant to high pH and/or high

temperature conditions. 1 and 2 Since enzymes produced by alkalophiles are active in the alkaline pH range, they are found to be most suitable in detergent formulations. The search for new species of microbes having the ability to produce industrially important enzymes with novel properties is a continuous process. The aim of this study was to search for alkalophilic bacteria having the ability to produce two industrially important alkaline enzymes viz. alkaline protease and alkaline amylase. Looking to the increased demand of alkaline protease and alkaline amylase 3, 4 and 5 in detergent industry and in treatment of alkaline wastes, studies on the cost effective production of these enzymes Selleck GSK126 is essential. Multiple enzymes produced from a single organism can be a useful step in this direction. 6 The work undertaken deals with the concomitant production of alkaline protease and alkaline amylase by an alkalophilic bacterium viz. Bacillus agaradhaerens. This study focuses on phenotypic and phylogenetic analysis performed in order to establish the taxonomic position of the isolated strain of B. agaradhaerens. Alkalophilic bacteria were screened by enrichment culture technique from Z-VAD-FMK molecular weight diverse samples collected in and around the

city of Indore of Madhya Pradesh, India. These samples included soil, sewage and industrial effluents. The samples were inoculated in Horikoshi’s broth medium7 I, pH 10.0, containing (g %) glucose; 1.0, peptone; 0.5, yeast extract; 0.5, KH2PO4; 0.1, MgSO4; 0.02, Na2CO3 1.0 (separately sterilized), distilled water 100.0 ml, followed by isolation on Horikoshi’s agar medium these I (pH 10.0). Single colonies that developed after 48 h of incubation at 30 °C were isolated. The same medium was used for maintenance of the strains. The alkalophilic/alkalotolerant nature of isolates was determined by growing each isolate on

Horikoshi’s M-I (pH 7.0) agar medium and incubating at 30 °C for 24 h. Individual bacterial colonies obtained on Horikoshi’s M-I (pH 10.0) agar plates were evaluated for their proteolytic ability by measuring the zone of casein hydrolysis on milk agar medium, pH 10.0, containing (g %) peptone; 1.0, meat extract; 0.5, NaCl; 0.5, Na2CO3; 1.0, distilled water; 100.0 ml, agar; 2.0. Separately sterilized 10% skimmed milk and Na2CO3 were added to the sterilized nutrient agar base, cooled up to 45 °C. Likewise the amylolytic activity of the alkalophilic isolates was evaluated by measuring the zone of starch hydrolysis on starch agar medium, pH 10.0, containing (g %) starch; 2.0, peptone; 0.5, yeast extract; 0.1, KH2PO4; 0.2, MgSO4; 0.02, Na2CO3; 1.0, agar; 2.0, distilled water; 100.0 ml Na2CO3 was sterilized separately and mixed.