Overexpression and one‐step renaturation‐purification of the tagged creatinine deiminase of Corynebacterium glutamicum in Escherichia coli cells. (19th February 2020)

• Record Type: Journal Article
• Authors: Zakalskiy, Andriy E.; Stasyuk, Nataliya Ye.; Zakalska, Oksana M.; Boretsky, Yuriy R.; Gonchar, Mykhailo V.
• Is Part Of: Cell biology international. Volume 44:Number 5(2020)
• Page Start: 1204
• Page End: 1211
• Publication Date: 2020-02-19
• Subjects: Corynebacterium glutamicum -- creatinine deiminase -- Escherichia coli -- gene cloning -- inclusion bodies -- metal‐affinity chromatography; Cytology -- Periodicals; Cells -- Periodicals; 571.605
• DOI: 10.1002/cbin.11320
• Languages: English
• ISSNs: 1065-6995

Abstract

Abstract: 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.