Open AccessNucleotide Sequence of the -Amylase-Pullulanase Gene from Clostridium Thermohydrosulfuricum
Author(s) -
Hannes Melasniemi,
Marja Paloheimo,
Leena Hemiö
Publication year - 1990
Publication title -
journal of general microbiology/journal of general microbiology
Language(s) - English
Resource type - Journals
eISSN - 2059-9323
pISSN - 0022-1287
DOI - 10.1099/00221287-136-3-447
Subject(s) - nucleic acid sequence , biology , gene , peptide sequence , bacillus subtilis , open reading frame , pullulanase , biochemistry , ribosomal binding site , protein primary structure , geobacillus stearothermophilus , genetics , structural gene , amylase , sequence alignment , enzyme , ribosome , escherichia coli , thermophile , bacteria , rna
The nucleotide sequence of the gene (apu) encoding the thermostable alpha-amylase-pullulanase of Clostridium thermohydrosulfuricum was determined. An open reading frame of 4425 bp was present. The deduced polypeptide (Mr 165,600), including a 31 amino acid putative signal sequence, comprised 1475 amino acids, with no cysteine residues. The structural gene was preceded by the consensus promoter sequence TTGACA TATAAT, a putative regulatory sequence and a putative ribosome-binding sequence AAA. The codon usage resembled that of Bacillus genes. The deduced sequence of the mature apu product showed similarities to various amylolytic enzymes, especially the neopullulanase of Bacillus stearothermophilus, whereas the signal sequence showed similarity to those of the alpha-amylases of B. stearothermophilus and B. subtilis. Three regions thought to be highly conserved in the primary structure of alpha-amylases could also be distinguished in the apu product, two being partly 'duplicated' in this alpha-1,4/alpha-1,6-active enzyme.