Место издания:Innovations and High Technologies MSU Ltd Moscow
Первая страница:115
Последняя страница:115
Аннотация:Penicillin acylase (EC 3.5.1.11, PA) is heterodimer. Active enzyme is produceв from single chain protein precursor through three steps of post-translational modification. Complicated maturation of precursor results in long time cultivation of recombinant strain E. coli producing PA (about 72 hours).
Previously we reported about preparation of permuted single chain penicillin acylase from Al-caligenes faecalis VKM B-1518 (pAfPA1). It was created by order change in DNA of gene of nucleo-tide sequences coding alpha- and beta- subunits and introduction of short linker between subunits. Here we report about preparation of the second version of permuted AfPA (pAfPA2) which contained shorter linker. pAfPA2 was expressed in E. coli cells as active and soluble enzyme. It was purified by the same procedure as developed for preparation of pAfPA1. It was shown that pAfPA2 has practically the same catalytic properties as wild-type AfPA and pAfPA1. Thermal stability was characterized through study of thermal inactivation kinetics. It was the same as one for pAfPA1.
The work was supported by Russian Foundation for Basic Research (RFBR),
grant 13-04-01907-а.