Partial purification and characterization of L-asparaginase from an endophytic Talaromyces pinophilus isolated from the rhizomes of Curcuma amada

• First report on characterization of l-asparaginase from an endophytic fungus.
• The enzyme is stable over a good range of pH and temperature.
• The enzyme is a heterodimer of 61.9 kDa and 39.1 kDa.
• The enzyme has a turnover number of 286.26 s−1.
• Cysteine residues play critical role in catalysis.

l-Asparaginase is a commercially significant enzyme. There exists a demand for a broad variety of microbial l-asparaginases with characteristics compatible with its different applications. Endophytic microorganisms, in particular are emerging as potential sources of l-asparaginases.The current work involves partial purification and characterization of l-asparaginase from Talaromyces pinophilus, an endophytic fungus isolated from the rhizomes of Curcuma amada. Maximum enzyme activity could be achieved at pH 8.0 and with temperature 28 °C. The enzyme Exhibits 95 % and 98% of its total activity at physiological pH and temperature, respectively. The enzyme activity is largely unhindered in the presence of metal ions such as Ca2+, Cu2+, Fe2+, Mg2+, Mn2+, Zn2+. Increase in the enzyme activity in the presence of thiol groups and reduction in the same upon addition of thiol blockers indicates the involvement of cysteine in the enzyme’s catalytic activity. The enzyme is a heterodimer of 62 kDa and 39 kDa. The enzyme has a Km of 6.4 mM, its turnover number towards l-asparagine is 286.3 s−1. The enzyme has 16% glutaminase activity; its Km towards glutamine is 13.3 mM and turnover number is 54.6 s−1.Our results highlight that l-asparaginase from endophytic Talaromyces pinophilus could be considered as potential candidate for clinical and industrial trials, owing to its efficiency and biochemical properties. To the best of our knowledge, this is the first report on partial purification and characterization of L-asparaginase from an endophyte.

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