Cloning, high-level expression and enzymatic properties of an intracellular serine protease from Bacillus sp. WRD-2
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Title |
Cloning, high-level expression and enzymatic properties of an intracellular serine protease from Bacillus sp. WRD-2
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Creator |
An, Sun-Young
Ok, Min Kim, Ji-Youn Jang, Moon-Sun Cho, Young-Su Choi, Yong-Lark Kim, Cheorl-Ho Lee, Young-Choon |
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Subject |
Bacillus
Cloning Overexpression Intracellular serine protease Purification Recombinant enzyme |
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Description |
141-147
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. |
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Date |
2009-04-02T06:32:11Z
2009-04-02T06:32:11Z 2004-08 |
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Type |
Article
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Identifier |
0301-1208
http://hdl.handle.net/123456789/3711 |
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Language |
en_US
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Publisher |
CSIR
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Source |
IJBB Vol. 41(4) [August 2004]
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