Abstract:
Papain (EC 2.4.22.2) is a globular protein family consisting of a single polypeptide chain with
three disulfide bridges and sulfuhydryl group necessary for proteolytic activity; it is an enzyme
of Carica papaya. The aim of this study is to extract, purify and characterize papain enzyme (EC
2.4.22.2) from the leaves of Carica papaya. Papain enzymes from Carica papaya was purified in
a 3-step purification using NH4(SO4)2 (70%), DEAE-Cellulose column chromatography and
Sephadex G-25 column chromatography to achieve different purification folds, as different the
fractions of the various peaks were pooled together. The effect of temperature, pH, substrate
concentration and various concentrations of Mg2+ concentrations on different casein
concentration was determined on the activity of the enzyme in the various pools. The findings
from this study shows that purified papain enzyme from Carica papaya has optimum active from
50°C - 59°C and pH 4.5 – 6.6. Its activity is generally enhanced by addition of Mg2+. However,
the enzyme is best activated at a concentration of 10mM of Mg2+
. The Km values for the papain
enzymes ranges from 1.47mg/ml – 8.70mg/ml and the Vmax value of the enzyme ranges from
0.042µmol/ml/min – 0.4167µmol/ml/min. The values of the parameters obtained from this study
could be put to use by the industry in the production of papain and also for household and
medical use in storing the product.
