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Research Journal of Microbiology

Year: 2017 | Volume: 12 | Issue: 3 | Page No.: 187-201
DOI: 10.17311/jm.2017.187.201
Production and Partial Purification of Hyperthermostable Alkaline Amylase in a Newly Isolated Bacillus cereus (sm-sr14) from Hot-spring Water
Sumit Sahoo, Sudipta Roy and Smarajit Maiti

Abstract: Background and Objective: Thermostable amylases from bacteria are of great importance for industrial purposes and for today’s biotechnological applications. This research aimed to study industrially important and physico-chemical stress-resistant bacterial amylase and its applications. Methodology: Amylase producing bacteria was collected from Bakreshwar hot-spring water, India by serial-dilution plating method on starch agar media. The strain, sm-sr14 was selected due to its strong hydrolysis efficiency. The strain was identified by polyphasic approach using biochemical, physiological properties and 16S rDNA homolgy. The optimization for amylase production, enzyme characterization and production using solid-state natural substrate were carried out with special emphasis on its heat stability by DNS method. The SPSS 10 was used for statistical analysis. Results: A hyperthermostable (60% residual activity up to 250°C) and highly alkaline (8-13; optimum 10) amylase producing Bacillus cereus sm-sr14 (Accession No. KM251578.1) was obtained from hot spring water. The amylase activity was found to be optimum at 50°C for 30 min incubation. The divalent cations Mn2+ and Ca2+ played critical role by 56% increase of enzyme activity. The EDTA (3 mM) moderately inhibits enzyme activity whereas, it was increased by the same concentration of β-mercaptoethanol. Enzyme was partially purified by cold acetone precipitation. The molecular mass was found to be 81 kDa by SDS PAGE. This amylase was found to be highly catalytic towards natural solid substrates like potato, rice, wheat and corn which was almost similar to laboratory-grade purified soluble and insoluble starch. Conclusion: The present findings suggested isolation and characterization of a hyperthermostable alkaline amylase with some important biotechnological potential ranging from food, fermentation, textile, detergent to paper industries.

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How to cite this article
Sumit Sahoo, Sudipta Roy and Smarajit Maiti, 2017. Production and Partial Purification of Hyperthermostable Alkaline Amylase in a Newly Isolated Bacillus cereus (sm-sr14) from Hot-spring Water. Research Journal of Microbiology, 12: 187-201.

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