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The Modified Heparin-Binding l-Asparaginase of Wolinella succinogenes

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Abstract

The modified asparaginase Was79 was derived from the recombinant wild-type l-asparaginase of Wolinella succinogenes. The Was79 contains the amino acid substitutions V23Q and K24T responsible for the resistance to trypsinolysis and the N-terminal heparin-binding peptide KRKKKGKGLGKKR responsible for the binding to heparin and tumor K562 cells in vitro. When tested on a mouse model of Fischer lymphadenosis L5178Y, therapeutic efficacy of Was79 was significantly higher than that of reference enzymes at all single therapeutic doses used (125–8000 IU/kg). At Was79 single doses of 500–8000 IU/kg, the complete remission rate of 100 % was observed. The Was79 variant can be expressed intracellularly in E. coli as a less immunogenic formyl-methionine-free form at high per cell production levels.

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Acknowledgments

This work was supported by the Ministry of Education and Science of RF (State contract No. 14.N08.11.0014).

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Authors and Affiliations

  1. State Research Institute for Genetics and Selection of Industrial Microorganisms, Moscow, Russia, 117545

    E. P. Sannikova, N. V. Bulushova, S. E. Cheperegin, I. I. Gubaydullin, G. G. Chestukhina, V. V. Ryabichenko, I. A. Zalunin, E. K. Kotlova, G. E. Konstantinova, T. S. Kubasova, S. V. Yarotsky, B. D. Efremov & D. G. Kozlov

  2. N. N. Blokhin Russian Cancer Research Center, Kashirskoye Shosse 24, Moscow, Russia, 115478

    A. A. Shtil & V. S. Pokrovsky

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  1. E. P. Sannikova
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  2. N. V. Bulushova
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  3. S. E. Cheperegin
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Correspondence to D. G. Kozlov.

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All applicable international, national, and institutional guidelines for the care and use of animals were followed.

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Sannikova, E.P., Bulushova, N.V., Cheperegin, S.E. et al. The Modified Heparin-Binding l-Asparaginase of Wolinella succinogenes . Mol Biotechnol 58, 528–539 (2016). https://doi.org/10.1007/s12033-016-9950-1

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  • DOI: https://doi.org/10.1007/s12033-016-9950-1

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