|
ИСТИНА |
Войти в систему Регистрация |
ИСТИНА ПсковГУ |
||
Cloning, expression and characterization of the recombinant Yersinia pseudotuberculosis L-asparaginaseстатья
- Авторы: Pokrovskaya MV, Aleksandrova SS, Pokrovsky VS, Omeljanjuk NM, Borisova AA, Anisimova NY, Sokolov NN
- Журнал: Protein Expression and Purification
- Том: 82
- Номер: 1
- Год издания: 2012
- Издательство: Academic Press
- Местоположение издательства: United States
- DOI: 10.1016/j.pep.2011.12.005
- Аннотация: We have cloned ansB (YPTB1411) gene from Yersinia pseudotuberculosis Q66CJ2 and constructed stable inducible expression system that overproduce L-asparaginase from Y. pseudotuberculosis (YpA) in Escherichiacoli BL21 (DE3) cells. For purification of YpA we used Q-Sepharose and DEAE-Toyopearl column chromatography. We examined kinetics of the enzyme reaction, catalytic activity as a function of pH, temperature and ionic strength, thermostability and other enzyme properties. Biochemical properties of YpA are similar with those of E. coli type II L-asparaginase. K(m) for L-asparagine is 17 ± 0.9 μM and pI 5.4 ± 0.3. Enzyme demonstrates maximum activity at pH 8.0 and 60 °C. YpA L-glutaminase activity is relatively low and more than 15 times less than specific activity towards L-asn. We evaluated also the antiproliferative effect of YpA in vitro and in vivo with E. colil-asparaginase (EcA) as the reference substance at similar conditions.
- Добавил в систему: Покровский Вадим Сергеевич