Volume 5 Supplement 1
Heterologous overexpression of a halophilic α-amylase
© Bautista et al; licensee BioMed Central Ltd. 2006
Published: 10 October 2006
Extracellular hydrolytic enzymes such as α-amylases are widely used in diverse applications in different industrial areas. α-amylase (EC 126.96.36.199) is an important endo-type carbohydrase that hydrolyzes α-1,4 glycosidic linkages of D-glucose oligomers and polymers. This enzyme has been found in organisms of the three Domains, being a key enzyme of carbohydrate metabolism. Haloferax mediterranei is an extremely halophilic Archaea that requires high salt concentrations to grow. This microorganism is able to grow in a minimal medium with ammonium acetate as the only source of carbon and nitrogen.H. mediterranei shows α-amylase extracellular activity when grows in this minimal medium in the presence of starch. The main role of this enzyme is the starch metabolism in the extracellular medium, so a lot of microorganisms depend on amylases for survival .
Furthermore, four genes have been sequenced by primer walking, which match up with four proteins belonging to ABC maltose transport system.
The α-amylase gene from H. mediterranei has been cloned, sequenced and overexpressed. The recombinant protein has been obtained in large amounts and refolded with a high efficiency.
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