Cloning and expression of an α-amylase encoding gene from the hyperthermophilic archaebacterium Thermococcus hydrothermalis and biochemical characterisation of the recombinant enzyme

被引:48
作者
Lévêque, E
Haye, B
Belarbi, A
机构
[1] Europol Agro, Ctr Biotechnol, UFR Sci Reims, Lab Microbiol Gen & Mol, F-51687 Reims 2, France
[2] Europol Agro, Ctr Biotechnol, UFR Sci Reims, Biochim Lab, F-51687 Reims 2, France
关键词
hyperthermophilic archaeon; alpha-amylase; nucleotide sequence; Thermococcus hydrothermalis;
D O I
10.1016/S0378-1097(00)00117-8
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
An alpha-amylase encoding gene from the extremely thermophilic Archaea Thermococcus hydrothermalis was cloned and expressed in Escherichia coli. The encoded alpha-amylase possesses molecular characteristics specific to the Archaea, especially from Pyrococcus species, with biochemical characteristics of the alpha-amylases from Thermococcus. The gene is 1374 bp long and encodes a protein of 457 amino acids composed of a 22 amino acid putative signal peptide and a 435 amino acid mature protein (calculated molecular mass 49 236 Da). The T. hydrothermalis recombinant alpha-amylase is optimally active at 75-85 degrees C and at pH 5.0-5.5. (C) 2000 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.
引用
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页码:67 / 71
页数:5
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