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Indian Journal of Biochemistry and Biophysics (IJBB) >
IJBB Vol.41 [2004] >
IJBB Vol.41(4) [August 2004] >
| Title: | Cloning, high-level expression and enzymatic properties of an intracellular serine protease from Bacillus sp. WRD-2 |
| Authors: | An, Sun-Young
Ok, Min
Kim, Ji-Youn
Jang, Moon-Sun
Cho, Young-Su
Choi, Yong-Lark
Kim, Cheorl-Ho
Lee, Young-Choon |
| Keywords: | Bacillus
Cloning
Overexpression
Intracellular serine protease
Purification
Recombinant enzyme |
| Issue Date: | Aug-2004 |
| Publisher: | CSIR |
| Abstract: | A gene, isp-B, encoding an intracellular serine protease from a newly isolated Bacillus sp. WRD-2 was cloned and characterized. Nucleotide sequence analysis showed an open reading frame of 960 bp encoding a polypeptide comprised of 319 amino acids. The primary structure of the enzyme predicted the structural features characteristic of other intracellular serine proteases, including active sites, Ser, His and Asp, as well as no signal sequence. The predicted amino acid sequence showed more than 60% homology with the intracellular serine proteases from Bacillus species. When expressed in E. coli, the recombinant enzyme (rISP-B) was overproduced in the cytoplasm as soluble and active form. The purified enzyme was completely inhibited by phenylmethylsulfonyl fluoride, EDTA and antipain. The enzyme showed maximum activity at pH 8.0 and 45ºC. It was stable at pH from 7.5 to 11.0 and below 50ºC. |
| Page(s): | 141-147 |
| ISSN: | 0301-1208 |
| Source: | IJBB Vol.41(4) [August 2004]
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