Identification of archaeon-producing hyperthermophilic α-amylase and characterization of the α-amy.pdfVIP

Appl Microbiol Biotechnol (2008) 80:605–614 DOI 10.1007/s00253-008-1561-8 BIOTECHNOLOGICALLY RELEVANT ENZYMES AND PROTEINS Identification of archaeon-producing hyperthermophilic α-amylase and characterization of the α-amylase Shujun Wang Zhaoxin Lu Mingsheng Lu Song Qin Hongfei Liu Xiangyuan Deng Qian Lin Jianan Chen Received: 22 February 2008 /Revised: 29 May 2008 /Accepted: 30 May 2008 / Published online: 28 June 2008 # Springer-Verlag 2008 Abstract The extremely thermophilic anaerobic archaeon shown by the sodium dodecyl sulfate-polyacrylamide gel strain, HJ21, was isolated from a deep-sea hydrothermal electrophoresis. vent, could produce hyperthermophilic α-amylase, and later was identified as Thermococcus from morphological, Keywords Hyperthermophilic α-amylase . biochemical, and physiological characteristics and the 16S Thermococcus sp. Thermostability . Archaeon . ribosomal RNA gene sequence. The extracellular thermo- Identification . Characterization stable α-amylase produced by strain HJ21 exhibited maximal activity at pH 5.0. The enzyme was stable in a broad pH range from pH 5.0 to 9.0. The optimal Introduction temperature of α-amylase was observed at 95°C. The half-life of the enzyme was 5 h at 90°C. Over 40% and α-Amylase(EC 3.2.1.1), endo-acting enzyme that hydro- 30% of the enzyme activity remained after incubation at lyzes starch by cleaving α-1,4-glucosidic linkages at random 100°C for 2 and 3 h, respectively. The enzyme did not sites, is one of the most important commercial enzymes require Ca2+ for thermostability. This α-amylase gene was widely used in starch-processing, brewing, alcohol produc- cloned, and its nucleotide sequence displayed an open tion, te