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Purification and Characterization of RNA Allied Extracellular Tyrosinase from Aspergillus Species

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Abstract

Production of l-DOPA, an anti-Parkinson’s drug, using biological sources is widely studied in which tyrosinase is known to play a vital role. Tyrosinase is an omnipresent type 3 copper enzyme participating in many essential biological functions. Understanding properties of tyrosinase is essential for developing useful tyrosinase-based applications. Hence, extracellular tyrosinase from Aspergillus flavus UWFP 570 was purified using ammonium sulphate precipitation and DEAE ion exchange chromatography up to 8.3-fold. Purified protein was a riboprotein in nature containing significant amount of RNA which was confirmed colorimetrically and by electrophoresis. Removal of RNA reduced the activity and altered the conformation of tyrosinase as suggested by spectroflurometric results. Optimum pH and temperature of this 140 kDa protein were 7 and 40 °C, respectively. Copper sulphate and magnesium chloride enhanced the activity whereas in contrast FeCl3 inhibited the activity completely. Purified tyrosinase transformed l-tyrosine (5 mM) to l-DOPA within 5 h.

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Acknowledgments

Author S. A. Inamdar thanks UGC, New Delhi, for BSR meritorious fellowship. V. A. Bapat wishes to thank Indian National Science Academy, New Delhi, for senior scientist position. Ms. S. M. Joshi thanks DBT, New Delhi, for the grant of SRF. J. P. Jadhav thanks DBT, New Delhi, for IPLS program.

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Authors and Affiliations

  1. Department of Biotechnology, Shivaji University, Kolhapur, 416 004, India

    Shrirang Inamdar, Swati Joshi, Vishwas Bapat & Jyoti Jadhav

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  1. Shrirang Inamdar
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  2. Swati Joshi
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  3. Vishwas Bapat
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  4. Jyoti Jadhav
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Correspondence to Jyoti Jadhav.

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Inamdar, S., Joshi, S., Bapat, V. et al. Purification and Characterization of RNA Allied Extracellular Tyrosinase from Aspergillus Species. Appl Biochem Biotechnol 172, 1183–1193 (2014). https://doi.org/10.1007/s12010-013-0555-x

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  • DOI: https://doi.org/10.1007/s12010-013-0555-x

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