Cloning and expression of an alpha-amylase encoding gene from the hyperthermophilic archaebacterium Thermococcus hydrothermalis and biochemical characterisation of the recombinant enzyme.
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 49236 Da). The T. hydrothermalis recombinant alpha-amylase is optimally active at 75-85 degrees C and at pH 5.0-5.5.
Lévêque
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Laboratoire de Microbiologie Générale et Moléculaire, Europol'Agro, Centre des Biotechnologies, UFR Sciences de Reims, France.
Haye
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Belarbi
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D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D017422:Sequence Analysis, DNA; D013696:Temperature; D019712:Thermococcus / Q000201:enzymology* / Q000235:genetics*; D000516:alpha-Amylases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
D011994:Recombinant Proteins; D000516:alpha-Amylases