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Cloning, expression, and purification of Helicobacter pylori L-asparaginase

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Abstract

Asparaginase from Helicobacter pylori (HpA) has been cloned and expressed in E. coli cells. The recombinant strain stably expressed catalytically active HpA. Optimization of culturing and expression conditions resulted in the expression level of the recombinant enzyme amounting up to 6% of total protein of the producer strain. A method developed for HpA purification included a single chromatographic stage and provided more than 60%-yield of the active enzyme. Specific asparaginase activity was 92 U/mg of protein, whereas the rate of glutamine hydrolysis was just 8.3 × 10−3 U/mg, respectively. Data obtained indicate that due to low glutaminase specificity HpA may be employed as a non-toxic enzyme preparation for treatment of leukemia.

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Authors and Affiliations

  1. Orekhovich Institute of Biomedical Chemistry, Russian Academy of Medical Sciences, Moscow, 119121, Russia

    Yu. A. Gladilina, N. N. Sokolov & J. V. Krasotkina

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  1. Yu. A. Gladilina
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  2. N. N. Sokolov
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  3. J. V. Krasotkina
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Correspondence to J. V. Krasotkina.

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Original Russian Text © Yu.A. Gladilina, N.N. Sokolov, J.V. Krasotkina, 2009, published in Biomeditsinskaya Khimiya.

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Gladilina, Y.A., Sokolov, N.N. & Krasotkina, J.V. Cloning, expression, and purification of Helicobacter pylori L-asparaginase. Biochem. Moscow Suppl. Ser. B 3, 89–91 (2009). https://doi.org/10.1134/S1990750809010132

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  • DOI: https://doi.org/10.1134/S1990750809010132

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