1887

Abstract

A recombinant clone expressing an amylase was identified from an generated genomic library of the thermophilic, moderately halophilic, anaerobic bacterium by activity screening, and the gene encoding the enzyme was designated AmyA. The gene was 1545 bp long, and encoded a 515 residue protein composed of a 25 amino acid putative signal peptide and a 490 amino acid mature protein. It possessed the five consensus regions characteristic of the α-amylase family and showed the greatest homology to the group of α-amylases. The gene was expressed in as a hexahistidine-tagged enzyme and purified. The purified recombinant enzyme was optimally active at 65 °C in 5% (w/v) NaCl at pH 75, with significant activity retained in the presence of up to 25% (w/v) NaCl. It had a specific activity of 2232 U mg and required NaCl and CaCl for optimum activity and thermostability. The relatively high proportion of acidic amino acids typically observed for many enzymes from halophiles was absent in AmyA.

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2002-08-01
2020-01-28
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