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The DNA sequence of the 5' region of the Bacillus licheniformis a-amylase gene is reported. Comparison of the inferred amino acid sequence of the B. licheniformis a-amylase gene with that of the Bacillus amyloliquefaciens gene shows that whereas the amino acid sequences of the mature proteins have considerable homology, the sequences for the signal peptides are distinct.Several Bacillus species secrete commercially important amylases (1,4-a-D-glucan glucanohydrolase; EC 3.2.1.1). Alpha-amylase genes from Bacillus subtilis (19,20), Bacillus amyloliquefaciens (12), Bacillus coagulans (2), and Bacillus licheniformis (11) have been cloned and expressed in either Escherichia coli or B. subtilis, and recently the DNA sequences of amylase genes from B. subtilis (19) and B. amyloliquefaciens (13, 16) have been determined. We report here the nucleotide sequence of the 5' regulatory region of the B. licheniformis gene, including the region coding for the signal peptide, which is not present on the mature form of the enzyme, and compare it with the sequenc,e of the B. amyloliquefaciens gene.The a-amylase gene from B. licheniformis FDO2 had previously been cloned onto B. subtilis replicons and localized to a 3.5-kilobase (kb) EcoRI fragment of DNA on a recombinant plasmid pSA33 (11). A simplified restriction map of the DNA in the vicinity of the gene is shown in Fig. 1. Comparison of the restriction enzyme cleavage map of this gene with the published map for the amylase gene from B. coagulans suggested that these two genes were quite similar. The construction of genetic fusions of the B. coagulans gene to the P-lactamase gene of E. coli (2) suggested that the 5' region of the B. coagulans gene was located to the left of a central PstI site. By analogy, we anticipated that the regulatory region of the B. licheniformis gene would be located to the left of the central PstI site in the EcoRI insert of pSA33. The region surrounding this PstI site has been seqvenced by both chemic,al and dideoxy methods. For the chemical sequencing, the 1.1-kb EcoRI-PstI fragment and the 2.3-kb PstI fragment lying on either side of the central PstI site were subcloned into the E. coli plasmid pUC8 (18). The DNA was sequenced on both strands from the HindIII site in pUC8, using the sequencing method of Maxam and Gilbert (8). For the dideoxy sequencing (7), the 0.7-kb MboI fragment spanning the central PstI site of the insert in pSA33 was excised and cut with PstI, and the two MboI-PstI fragments subcloned into pUC8 were cut with BamHI and PstI. The two inserts were sequenced directly after cutting with SmaI, using primer and other reagents supplied by Bethesda Research Laboratories. The portion of the sequence which includes the putative promoter, ribosome binding site, and NH2-terminal 75 amino acids is shown in Fig. 2. * Corresponding author.To the right of the central PstI site, an open reading frame was observed that would encode a protein with a sequence highly hormologous to the first 18 amino acids of the reported sequence of the mature pr...

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