Purification and characterization of Natural Solid Substrate Degrading and Alcohol Producing Hyperthermostable Alkaline Amylase from Bacillus cereus (sm-sr14).

Amylases enzymes hydrolyze starch molecules to producediverse products including dextrins, and progressively smaller polymers. These include glucose units linked through α-1-1, α-1-4, α-1-6, glycosidic bonds. This enzyme carry an (α /β) 8 or TIM barrel structure ia also produced containing the catalytic site residues.These groups of enzymespossess four conserved regions in their primary sequence. In the carbohydrate-degrading enzyme(CAZy) database, α-amylases are classified into different glycoside hydrolase families (GHF) based on their amino acid sequence. Present objectives were to study on one such enzyme based on its molecular characterization after it purification in our laboratory. Its main property of solid-natural starch degradation was extensively investigated for its pharmaceutical/ industrialapplications.Amylase producing bacteria Bacillus cereus sm-sr14 (Accession no. KM251578.1) was purified to homogeneity on a Seralose 6B-150 gel-matrix and gave a single peak during HPLC. MALDI-TOF mass-spectrometry with bioinformatics studies revealed its significant similarity to α/β hydrolase family. The enzyme showed a very high application-favourable Km,Vmax and Kcat during the catalysis of different natural solid starch materials. Analysis for hydrolytic product showed that this enzyme can be classified as the exoamylaseas because it produced significant amount of glucose.Beside the purified enzyme, the present whole organism Bacillus cereus sm-sr14 could degrade natural solid starch materials like potato, rice up to the application level in pharmaceutical/ industrial field for alcohol production.