Purification and biochemical characterisation of acid phosphatase-I from seeds of Nelumbo nucifera.
Nat Prod Res
; 30(5): 570-3, 2016.
Article
in En
| MEDLINE
| ID: mdl-25887488
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 Zn(2+), W(2+); weakly inhibited by Cu(2+), Mo(2+) and Cr(6+) and moderately activated by Mg(2+). 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.
Key words
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Seeds
/
Acid Phosphatase
/
Nelumbo
Language:
En
Journal:
Nat Prod Res
Year:
2016
Document type:
Article
Affiliation country: