Enhancement of anti-tumor activity in melanoma using arginine deiminase fused with 30Kc19α protein

Haein Lee, Geunhwa Park, Seulha Kim, Boram Son, Jinmyoung Joo, Hee Ho Park, Tai Hyun Park

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1 Scopus citations


Abstract: Arginine deiminase (ADI) is a microbial-derived enzyme which catalyzes the conversion of l-arginine into l-citrulline. ADI originating from Mycoplasma has been reported to present anti-tumor activity against arginine-auxotrophic tumors, including melanoma. Melanoma cells are sensitive to arginine depletion due to reduced expression of argininosuccinate synthase 1 (ASS1), a key enzyme for arginine biosynthesis. However, clinical applications of recombinant ADI for melanoma treatment present some limitations. Since recombinant ADI is not human-derived, it shows instability, proteolytic degradation, and antigenicity in human serum. In addition, there is a problem of drug resistance issue due to the intracellular expression of once-silenced ASS1. Moreover, recombinant ADI proteins are mainly expressed as inclusion body forms in Escherichia coli and require a time-consuming refolding process to turn them back into active form. Herein, we propose fusion of recombinant ADI from Mycoplasma hominis and 30Kc19α, a cell-penetrating protein which also increases stability and soluble expression of cargo proteins, to overcome these problems. We inserted matrix metalloproteinase-2 cleavable linker between ADI and 30Kc19α to increase enzyme activity in melanoma cells. Compared to ADI, ADI-LK-30Kc19α showed enhanced solubility, stability, and cell penetration. The fusion protein demonstrated selective cytotoxicity and reduced drug resistance in melanoma cells, thus would be a promising strategy for the improved efficacy in melanoma treatment. Key points: • Fusion of ADI with 30Kc19α enhances soluble expression and productivity of recombinant ADI in E.

Original languageEnglish
Pages (from-to)7531-7545
Number of pages15
JournalApplied Microbiology and Biotechnology
Issue number22
StatePublished - Nov 2022

Bibliographical note

Funding Information:
This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. 2020R1A4A3078645, 2021R1C1C1014606).

Publisher Copyright:
© 2022, The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.


  • Arginine deiminase (ADI)
  • Cell-penetrating protein
  • Enzyme stabilizer
  • Melanoma
  • Solubility enhancer


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