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PEGylated recombinant L-asparaginase from Erwinia carotovora: Production,properties, and potential applications
Authors:N. S. Melik-Nubarov  I. D. Grozdova  G. Yu. Lomakina  M. V. Pokrovskaya  V. S. Pokrovski  S. S. Aleksandrova  O. Yu. Abakumova  O. V. Podobed  D. V. Grishin  N. N. Sokolov
Affiliation:1.Department of Chemistry,Lomonosov Moscow State University,Moscow,Russia;2.Orekhovich Scientific Research Institute of Biomedical Chemistry,Moscow,Russia
Abstract:N-hydroxysuccinimide ester of monomethoxy polyethylene glycol hemisuccinate was synthesized. It acylated amino groups in a molecule of recombinant L-asparaginase from Erwinia carotovora. A method of L-asparaginase modification by the obtained activated polyethylene glycol derivative was developed. The best results were produced by modification of the enzyme with a 25-fold excess of reagent relative to the enzyme tetramer. The modified L-asparaginase was isolated from the reaction mixture by gel filtration on Sepharose CL-6B. The purified bioconjugate did not contain PEG unbound to the protein, demonstrated high catalytic activity, and exhibited antiproliferative action on cell cultures.
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