Ukr.Biochem.J. 2016; Volume 88, Issue 3, May-Jun, pp. 18-28

doi: https://doi.org/10.15407/ubj88.03.018

Isolation and purification of Bacillus thuringiensis var. israelensis IМV В-7465 peptidase with specificity toward elastin and collagen

21.06.2016   Uncategorized   No comments

N. А. Nidialkova1, L. D. Varbanets1, V. O. Chernyshenko2

1Institute of Microbiology and Virology, National Academy
of Sciences of Ukraine, Kyiv;
e-mail: Nidialkova@gmail.com;
2Palladin Institute of Biochemistry, National Academy
of Sciences of Ukraine, Kyiv

Peptidase of Bacillus thuringiensis var. israelensis IМV В-7465 was isolated from culture supernatant using consecutive fractionations by an ammonium sulphate (60% saturation), ion-exchange chromatography and gel-filtration on the TSK-gels Toyoperl HW-55 and DEAE 650(M). Specific elastase (442 U∙mg of protein-1) and collagenase (212.7 U∙mg of protein-1) activities of the purified enzyme preparation were 8.0- and 6.1-fold, respectively higher than ones of the culture supernatant. Peptidase yields were 33.5% for elastase activity and 30.1% for collagenase activity. It was established that the enzyme is serine metal-dependent alkaline peptidase with Mr about 37 kDa. Maximal hydrolysis of elastin and collagen occurs at the optimum pH 8.0 and t° – 40 and 50 °С, respectively. The purified preparation has high stability at pH in the range of 7.0 to 10.0 and 40-50 °С.

Keywords: , , , ,

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