Purification and biochemical characterisation of acid phosphatase-I from seeds of Nelumbo nucifera |
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| Authors: | Sanaullah Khan Shahnaz Khan Sajida Batool Mushtaq Ahmed |
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| Institution: | 1. Department of Bioscience, COMSATS Institute of Information Technology Islamabad, Islamabad, Pakistansanaullah.khan@comsats.edu.pk;3. Department of Chemistry, University of Science and Technology, Bannu, Pakistan;4. Department of Bioscience, COMSATS Institute of Information Technology Islamabad, Islamabad, Pakistan;5. Department of Biotechnology, University of Science and Technology, Bannu, Pakistan |
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| Abstract: | Acid phosphatase-I (Apase-I) from seeds of Nelumbo nucifera was purified to electrophoretic homogeneity by combination of ammonium sulfate precipitation, size-exclusion and ion exchange chromatography. SDS-PAGE of purified Apase-I gave a single band with molecular mass of 80 kDa under reducing and non-reducing conditions, indicating that the enzyme was a monomer. The purified enzyme showed maximum activity at 50°C and at pH 5. The Km, Vmax and Kcat for p-nitrophenyl phosphate were 132 μM, 10 μmol/min/mg and 6.7/sec respectively. Apase-I activity was strongly inhibited by Zn2+, W2+; weakly inhibited by Cu2+, Mo2+ and Cr6+ and moderately activated by Mg2+. The enzyme was shown to be thermolabile as it lost 50% of its activity at 50°C after incubation for 1 hour. The amino acid analysis of enzyme revealed high proportion of acidic amino acids, which is very similar to that of tomato Apase-I and lower than potato Apase. |
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| Keywords: | Nelumbo nucifera seeds acid phosphatase-I purification biochemical characterisation |
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