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Balabanova LA, Golotin VA, Bakunina IY, et al. Recombinant α-NAcetylgalactosaminidase from Marine Bacterium-Modifying A Erythrocyte Antigens. Acta Naturae. 2015;7(1):117-20
Balabanova, L. A., Golotin, V. A., Bakunina, I. Y., Slepchenko, L. V., Isakov, V. V., Podvolotskaya, A. B., & Rasskazov, V. A. (2015). Recombinant α-NAcetylgalactosaminidase from Marine Bacterium-Modifying A Erythrocyte Antigens. Acta naturae, 7(1), 117-20.
Balabanova, L A, et al. "Recombinant α-NAcetylgalactosaminidase from Marine Bacterium-Modifying A Erythrocyte Antigens." Acta naturae vol. 7,1 (2015): 117-20.
Balabanova LA, Golotin VA, Bakunina IY, Slepchenko LV, Isakov VV, Podvolotskaya AB, Rasskazov VA. Recombinant α-NAcetylgalactosaminidase from Marine Bacterium-Modifying A Erythrocyte Antigens. Acta Naturae. 2015 Jan-Mar;7(1):117-20. PMID: 25927009; PMCID: PMC4410403.
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Acta Naturae. 2015 Jan-Mar;7(1):117-20.

Recombinant α-NAcetylgalactosaminidase from Marine Bacterium-Modifying A Erythrocyte Antigens.

Acta naturae

L A Balabanova, V A Golotin, I Y Bakunina, L V Slepchenko, V V Isakov, A B Podvolotskaya, V A Rasskazov

Affiliations

  1. G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences, 100-letiya Vladivostoka Ave., 159, 690022, Vladivostok, Russia ; Far Eastern Federal University, Sukhanova Str., 8, 690950, Vladivostok, Russia.
  2. G.B. Elyakov Pacific Institute of Bioorganic Chemistry, Far Eastern Branch, Russian Academy of Sciences, 100-letiya Vladivostoka Ave., 159, 690022, Vladivostok, Russia.
  3. Far Eastern Federal University, Sukhanova Str., 8, 690950, Vladivostok, Russia.

PMID: 25927009 PMCID: PMC4410403

Abstract

A plasmid based on pET-40b was constructed to synthesize recombinant α-N-acetylgalactosaminidase of the marine bacterium Arenibacter latericius KMM 426T (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.

Keywords: 1H NMR spectroscopy; Arenibacter latericius; conversion of A erythrocytes; glycoside hydrolase GH109

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