Signal peptide of Bacillus subtilis alpha-amylase.
Mature alpha-amylase of Bacillus subtilis is known to be formed from its precursor by the removal of the NH2-terminal 41 amino acid sequence (41 amino acid leader sequence). DNA fragments coding for short sequences consisting of 28 (Pro as the COOH terminus) 29 (Ala), 31 (Ala), and 33 (Ala) amino acids from the translation initiator, Met, in the leader sequence were prepared and fused in frame to the DNA encoding the mature alpha-amylase. The secretion activity of the 33 amino acid sequence was nearly twice as high as that of the parental 41 amino acid sequence, whereas the activity of the 31 amino acid sequence was 75% of that of the parent. In contrast, almost no secretion activity was observed with the 28 and 29 amino acid sequences. The signal peptide cleavage site of the precursor expressed from the plasmid encoding the 33 amino acid sequence was located between Ala and Leu at positions 33 and 34 and that from the 31 amino acid sequence between Thr and Ala at positions 33 and 34. The NH2-terminal amino acid from the latter corresponded to the 3rd amino acid of the mature enzyme. These results indicated that the functional signal peptide of the B. subtilis beta-amylase consists of the first 33 amino acids from the initiator, Met.
Sasamoto
H
H
Institute of Biological Sciences, University of Tsukuba, Ibaraki.
Nakazawa
K
K
Tsutsumi
K
K
Takase
K
K
Yamane
K
K
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000201:enzymology; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D005110:Extracellular Space / Q000201:enzymology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010957:Plasmids; D021382:Protein Sorting Signals / Q000235:genetics / Q000378:metabolism*; D000516:alpha-Amylases / Q000096:biosynthesis* / Q000235:genetics
D004269:DNA, Bacterial; D021382:Protein Sorting Signals; D000516:alpha-Amylases