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Arginine deaminase from Pseudomonas aeruginosa PS2: purification, biochemical characterization and in-vitro evaluation of anticancer activity

3 Biotech volume 10, Article number: 226 (2020) Cite this article

Abstract

In the present study, arginine deaminase (ADI) was purified from Pseudomonas aeruginosa PS2 which showed relative molecular mass of 70 ± 3 kDa on native-PAGE and 36 ± 0.5 kDa on SDS-PAGE. Purified ADI exhibited optimum activity at pH 6.5 and temperature 40 ºC. Metal ions, K+ and Mg2+ had positive, while Mn2+, Cr2+, Co2+, Fe3+, Ni2+, Cu2+, Cd2+ and Hg2+ had negative effects on catalytic activity of ADI. Purified enzyme showed high substrate specificity towards natural substrate L-arginine and did not hydrolyse its structural analogues. In-vitro serum half-life of purified ADI was 40 h, whereas proteolytic half-life was 28, 27, and 32 min against trypsin, elastase-I and proteinase-K, respectively. Anticancer activity of ADI has been evaluated against panel of human cancer cell lines (LS-180, HCT-116, MCF-7, BT-549, T47D, HL-60, MOLT-4, K-562, and PC-3) but lowest IC50 1.2 IU ml−1 was recorded with MCF-7 cells. Colony forming assay, wound-healing migration assay, phase contrast microscopy, DAPI staining, cell cycle analysis and DNA laddering assay revealed that ADI treatment induced apoptotic cell death in dose dependent manner. Increased level of MMP loss, ROS generation and decreased level of SOD, CAT, GPx and GSH displayed ADI treatment induced mitochondrial dysfunctioning. Furthermore, purified ADI had no substantial toxicity against human normal cell lines and blood erythrocytes. These findings suggesting that purified ADI could be developed as an anticancer agent but more in depth studies are warranted.

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Acknowledgements

Authors are thankful to the Head, Department of P.G. Studies and Research in Biological Science, Rani Durgavati University, Jabalpur, Madhya Pradesh, India, for providing Laboratory facilities to carry out this work. Islam Husain is thankful to Department of Science and Technology (DST), Indian, for providing N-PDF Grant (DST No: PDF/2016/002697).

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  1. Bacteriology Laboratory, Department of P.G. Studies and Research in Biological Science, Rani Durgavati University, Jabalpur, Madhya Pradesh, India

    Kiran Bala, Islam Husain & Anjana Sharma

  2. Molecular and Structural Biology Division, CSIR-Central Drug Research Institute, Sector 10, Jankipuram Extension, Lucknow, Uttar Pradesh, India

    Islam Husain

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  1. Kiran Bala
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  2. Islam Husain
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Correspondence to Anjana Sharma.

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Bala, K., Husain, I. & Sharma, A. Arginine deaminase from Pseudomonas aeruginosa PS2: purification, biochemical characterization and in-vitro evaluation of anticancer activity. 3 Biotech 10, 226 (2020). https://doi.org/10.1007/s13205-020-02212-6

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  • DOI: https://doi.org/10.1007/s13205-020-02212-6

Keywords

  • Arginine deiminase
  • Anticancer activity
  • Antioxidants
  • MCF-7
  • Hemolysis