Open AccessA few amino acid substitutions are responsible for the higher thermostability of a novel NAD + ‐dependent bacillar alcohol dehydrogenase
Author(s) -
CANNIO Raffaele,
ROSSI Mosè,
BARTOLUCCI Simonetta
Publication year - 1994
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1994.tb18873.x
Subject(s) - thermostability , sulfolobus solfataricus , thermophile , alcohol dehydrogenase , biochemistry , biology , amino acid , peptide sequence , nucleic acid sequence , nad+ kinase , escherichia coli , gene , enzyme , microbiology and biotechnology , archaea
The gene adh‐hT encoding a thermostable and thermophilic NAD + ‐dependent alcohol dehydrogenase (ADH) from the novel and more thermophilic Bacillus stearothermophilus LLD‐R strain was cloned and its nucleotide sequence determined. The deduced protein sequence shows remarkable amino acid substitutions when compared to the sequence of the protein isolated from strain NCA1503 and significant similarity with the highly thermostable ADH from the thermoacidophilic archaebacterium Sulfolobus solfataricus . The alignment of these sequences led to the identification of three amino acid replacements probably responsible for the higher thermostability of the novel bacillar ADH. Adh‐hT gene expression in Escherichia coli , a fast purification procedure and the characterization of the recombinant enzyme are also described.