The Modified Heparin-Binding l-Asparaginase of Wolinella succinogenes

• Published in: Molecular biotechnology 2016-09, Vol.58 (8-9), p.528-539
• Main Authors: Sannikova, E. P., Bulushova, N. V., Cheperegin, S. E., Gubaydullin, I. I., Chestukhina, G. G., Ryabichenko, V. V., Zalunin, I. A., Kotlova, E. K., Konstantinova, G. E., Kubasova, T. S., Shtil, A. A., Pokrovsky, V. S., Yarotsky, S. V., Efremov, B. D., Kozlov, D. G.
• Format: Article
• Language: English
• Subjects: Amino Acid Substitution; Amino acids; Animals; Antineoplastic Agents - administration & dosage; Antineoplastic Agents - pharmacology; Asparaginase - administration & dosage; Asparaginase - genetics; Asparaginase - metabolism; Asparaginase - pharmacology; Bacterial Proteins - administration & dosage; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Bacterial Proteins - pharmacology; Biochemistry; Biological Techniques; Biotechnology; Cell Biology; Cell Proliferation - drug effects; Cell Survival - drug effects; Chemistry; Chemistry and Materials Science; E coli; Enzymes; Escherichia coli; Gram-negative bacteria; Heparin - metabolism; Human Genetics; Humans; K562 Cells; Leukemia L5178 - drug therapy; Mice; Mutagenesis; Original Paper; Peptides; Protein Science; Recombinant Proteins - metabolism; Recombinant Proteins - pharmacology; Wolinella - enzymology; Wolinella - genetics; Wolinella succinogenes; Xenograft Model Antitumor Assays
• ISSN: 1073-6085; 1559-0305
• DOI: 10.1007/s12033-016-9950-1

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.