Overexpression and one-step renaturation-purification of the tagged creatinine deiminase of Corynebacterium glutamicum in Escherichia coli cells.

Autor: Zakalskiy AE; Institute of Cell Biology, National Academy of Sciences of Ukraine, 14/16 Drahomanov Str., 79005, Lviv, Ukraine.; Ivan Bobersky Lviv State University of Physical Culture, 11 Kostiushko Str., 79000, Lviv, Ukraine., Stasyuk NY; Institute of Cell Biology, National Academy of Sciences of Ukraine, 14/16 Drahomanov Str., 79005, Lviv, Ukraine., Zakalska OM; Institute of Cell Biology, National Academy of Sciences of Ukraine, 14/16 Drahomanov Str., 79005, Lviv, Ukraine., Boretsky YR; Ivan Bobersky Lviv State University of Physical Culture, 11 Kostiushko Str., 79000, Lviv, Ukraine., Gonchar MV; Institute of Cell Biology, National Academy of Sciences of Ukraine, 14/16 Drahomanov Str., 79005, Lviv, Ukraine.; Ivan Bobersky Lviv State University of Physical Culture, 11 Kostiushko Str., 79000, Lviv, Ukraine.; Drohobych Ivan Franko State Pedagogical University, 24 Ivan Franko Str., 82100, Drohobych, Ukraine.
Jazyk: angličtina
Zdroj: Cell biology international [Cell Biol Int] 2020 May; Vol. 44 (5), pp. 1204-1211. Date of Electronic Publication: 2020 Feb 19.
DOI: 10.1002/cbin.11320
Abstrakt: The codA gene of Corynebacterium glutamicum PCM 1945 coding for a creatinine deiminase (CDI) (EC 3.5.4.21) has been amplified and cloned. The recombinant strain of Escherichia coli that overproduces the (His) 6 -tagged inactive CDI of C. glutamicum as inclusion bodies has been constructed. After solubilization of inclusion bodies in the presence of 0.3% N-lauroylsarcosine, the enzyme was renaturated and purified by a single-step procedure using metal-affinity chromatography. The yield of the (His) 6 -tagged CDI is ~30 mg from 1 L culture. The purified enzyme is sufficiently stable under the conditions designed and possesses an activity of 10-20 U/mg. The main characteristics of the tagged enzyme remained similar to that of the natural enzyme.
(© 2020 International Federation for Cell Biology.)
Databáze: MEDLINE
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