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Extracellular urease from Arthrobacter creatinolyticus MTCC 5604: scale up, purification and its cytotoxic effect thereof

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Abstract

Urease is a potent metalloenzyme with diverse applications. This paper describes the scale up and purification of an extracellular urease from Arthrobacter creatinolyticus MTCC 5604. The urease production was scaled-up in 3.7 L and 20 L fermentor. A maximum activity of 27 and 27.8 U/mL and a productivity of 0.90 and 0.99 U/mL/h were obtained at 30 h and 28 h in 3.7 and 20 L fermentor, respectively. Urease was purified to homogeneity with 49.85-fold purification by gel filtration and anion exchange chromatography with a yield of 36% and a specific activity of 1044.37 U/mg protein. The enzyme showed three protein bands with molecular mass of 72.6, 11.2 and 6.1 kDa on SDS-PAGE and ~ 270 kDa on native PAGE. The cytotoxic effect of urease was assessed in vitro using cancer cell lines (A549 and MG-63) and normal cell line (HEK 293). Urease showed its inhibitory effects on cancer cell lines through the generation of toxic ammonia, which in turn increased the pH of the surrounding medium. This increase in extracellular pH, enhanced the cytotoxic effect of weak base chemotherapeutic drugs, doxorubicin (50 µM) and vinblastine (100 µM) in the presence of urease (5 U/mL) and urea (0–4 mM) significantly.

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Acknowledgements

The authors thank the Director, CSIR - Central Leather Research Institute, Chennai, India, for his kind permission to publish this work. The financial assistance rendered by University Grants Commission and DST - INSPIRE, Government of India is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Biochemistry and Biotechnology, CSIR - Central Leather Research Institute, Adyar, Chennai, 600 020, India

    Ramesh Rajendran, Ajitha Pandi, Aparna Ramchary, Hemalatha Thiagarajan, Ayyadurai Niraikulam, Gowthaman Marichetti Kuppuswami & Kamini Numbi Ramudu

  2. Department of Pharmaceutical Technology, International Medical University, Bukit Jalil, 57000, Kuala Lumpur, Malaysia

    Jithendra Panneerselvam

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  1. Ramesh Rajendran
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  2. Ajitha Pandi
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Correspondence to Kamini Numbi Ramudu.

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Rajendran, R., Pandi, A., Ramchary, A. et al. Extracellular urease from Arthrobacter creatinolyticus MTCC 5604: scale up, purification and its cytotoxic effect thereof. Mol Biol Rep 46, 133–141 (2019). https://doi.org/10.1007/s11033-018-4453-8

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  • DOI: https://doi.org/10.1007/s11033-018-4453-8

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