|
KMID : 0545119940040020113
|
|
Journal of Microbiology and Biotechnology
1994 Volume.4 No. 2 p.113 ~ p.118
|
|
|
Purification and Characterization of a Thermostable Protease from Pseudomonas aeruginosa NS-83
|
|
Kim, Hyung Kwoun
Kim, Kee Hyun/Lee, Jung Kee/Bae, Kyung Sook/Sung, Chang/Oh, Tae Kwang
|
|
|
Abstract
|
|
|
|
A bacterial strain NS-83 isolated from soil was able to produce an extracellular thermostable protease. The strain was identified as Pseudomonas aeruginosa based on its morphological and physiological characteristics. A thermostable protease from this strain has been purified to homogeneity as judged by SDS-PAGE and isoelectric focusing. The purification procedures included hydrophobic interaction, ion exchange, and gel filtration chromatography. The M_r and the pl of the enzyme were 32,000 and 5.9, respectively. The optimal pH at 55℃ and the optimal temperature at pH 7.0 were 8.0 and 60℃, respectively. The D-values of the enzyme at 60, 65, and 70℃ were 22, 2.1, and 0.75 hrs, respectively. The enzyme activity was significantly inhibited in the presence of 1 mM o-phenanthroline or EDTA, suggesting that the enzyme is metalloprotease. The K_m and V_max for Hammarsten casein were found to be 3.2 ㎎/㎖ and 0.918 unit/㎖, respectively. These enzymatic properties were similar to those of elastase produced from P. aeruginosa IFO 3455, but the enzyme was clearly different from the reported elastase, in respect to Ca^++ effects on enzyme-thermostability. This property, together with amino acid composition analysis, confirmed that the enzyme differs from the known P. aeruginosa elastase.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
  
|
|