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 22.214.171.124) 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.
- Jones RA, Jermiin LS, Easteal S, Patel BKC, Beacham IR: Amylase and 16S rRNA genes from a hyperthermophilic archaebacterium. J Appl Microbiol. 1999, 86: 93-107. 10.1046/j.1365-2672.1999.00642.x.View ArticleGoogle Scholar
- Pérez-Pomares F, Bautista V, Ferrer J, Pire C, Marhuenda-Egea FC, Bonete MJ: Alpha-amylase activity from the halophilic archaeon Haloferax mediterranei . Extremophiles. 2003, 7 (4): 299-306. 10.1007/s00792-003-0327-6.View ArticleGoogle Scholar
- Pire C, Esclapez J, Ferrer J, Bonete MJ: Heterologous overexpression of glucose dehydrogenase from the halophilic archaeon Haloferax mediterranei, an enzyme of the medium chain dehydrogenase family. FEMS Microbiol Lett. 2001, 200: 221-227. 10.1111/j.1574-6968.2001.tb10719.x.View ArticleGoogle Scholar
This article is published under license to BioMed Central Ltd.