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KMID : 1007519970060030224
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Food Science and Biotechnology
1997 Volume.6 No. 3 p.224 ~ p.228
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Cloning and Molecular Characteristics of an apr Gene of Bacillus subtilis NS15-4 Encoding an Alkaline Protease
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OH, TAE KWANG
Nam, Hee Sop/Lee, Hyung Jae/Kim, Jong Hwa/Chae, Keon Sang/Jeong, Hyo Young/Yang, Moon Sik
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Abstract
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Three independently amplified DNA fragments of an aprNS gene of Bacillus subtilis NS15-4 encoding an alkaline protease were obtained by polymerase chain reaction (PCR) and their nucleotide sequences were analyzed in order to find out the correct nucleotide sequences of the gene. Nucleotide sequence of one fragment, which is 1,389 bp in a total, was determined completely in both directions, and those of the other two were done in one direction, partially in both directions. The determined nucleotide sequences indicated that there is an 1,143 by open reading frame (ORF) possibly encoding a 381 amino acid polypeptide. A putative promoter sequence possibly acting in Escherichia coli and a Shine-Dalgarno (SD) sequence were found in base pairs between 29 and 60, and 9 bp upstream to the translation initiation codon, GTG, respectively. The aprNS gene showed a similarity with that for subtilisin Amylosacchariticus (94%), subtilisin Carlsberg (94%), or subtilisin NAT (93%) in the nucleotide sequence. The deduced amino cid sequence revealed a putative N-terminal signal peptide of 29 residues and a pro-peptide of 77 residues followed by the 275 amino acid-mature protein. The AprNS ORF had codon bias index (CBI) of 0.2, suggesting that it may not be translated in a large amount in E. coli, and showed 95% similarity with subtilisin NAT, subtilisin J, and subtilisin Amylosacchariticus in the amino acid sequence.
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KEYWORD
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