Email this article Recombinant α-NAcetylgalactosaminidase from Marine Bacterium-Modifying A Erythrocyte Antigens
- Authors: Balabanova L.A.1,2, Golotin V.А.1, Bakunina I.Y.1, Slepchenko L.V.1, Isakov V.V1, Podvolotskaya A.B.2, Rasskazov V.А.1
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Affiliations:
- G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences
- Far Eastern Federal University
- Issue: Vol 7, No 1 (2015)
- Pages: 117-120
- Section: Short communications
- URL: https://actanaturae.ru/2075-8251/article/view/10520
- DOI: https://doi.org/10.32607/20758251-2015-7-1-117-120
- ID: 10520
Cite item
Abstract
A plasmid based on pET-40b was constructed to synthesize recombinant α-N-acetylgalactosaminidase of the marine bacterium Arenibacter latericius KMM 426 T (α-AlNaGal) in Escherichia coli cells. The yield of α-Al-NaGal attains 10 mg/ml with activity of 49.7 ± 1.3 U at 16°C, concentration of inductor 2 mM, and cultivation for 12 h. Techniques such as anion exchange, metal affinity and gel filtration chromatography to purify α-AlNaGal were applied. α-AlNaGal is a homodimer with a molecular weight of 164 kDa. This enzyme is stable at up to 50°C with a temperature range optimum activity of 20-37°C. Furthermore, its activity is independent of the presence of metal ions in the incubation medium. 1H NMR spectroscopy revealed that α-AlNaGal catalyzes the hydrolysis of the O-glycosidic bond with retention of anomeric stereochemistry and possesses a mechanism of action identical to that of other glycoside hydrolases of the 109 family. α-AlNaGal reduces the serological activity of A erythrocytes at pH 7.3. This property of α-AlNaGal can potentially be used for enzymatic conversion of A and AB erythrocytes to blood group O erythrocytes.
About the authors
L. A. Balabanova
G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences; Far Eastern Federal University
Author for correspondence.
Email: balaban@piboc.dvo.ru
Russian Federation
V. А. Golotin
G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences
Email: balaban@piboc.dvo.ru
Russian Federation
I. Yu. Bakunina
G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences
Email: balaban@piboc.dvo.ru
Russian Federation
L. V. Slepchenko
G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences
Email: balaban@piboc.dvo.ru
Russian Federation
V. V Isakov
G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences
Email: balaban@piboc.dvo.ru
Russian Federation
A. B. Podvolotskaya
Far Eastern Federal University
Email: balaban@piboc.dvo.ru
Russian Federation
V. А. Rasskazov
G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences
Email: balaban@piboc.dvo.ru
Russian Federation
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