PROTEASES FROM CELL CULTURE OF Bromelia hemisphaerica ...

Food Science and Biotechnology for Developing Countries.2004
U/m g
6.4
5.6
4.8
4.0
3.2
2.4
1.6
0.8
0.0
30 35 40 45 50 55 60
T (°C)
65 70 75 80 85 90
Fig. 5. Temperature effect on P-III enzyme activity with 1% casein- 0.05 M phosphate buffer,
pH 7.6 and 20 mM Cys at 35 °C.
The above values are in the range of optimal pH and temperature reported for
another cysteine proteases of plants and are similar to that from another proteases
isolated from the same latex source. The studies of pH stability for P-III showed that
this enzyme maintains 100% of proteolytic activity within a 2-24 hs period along the
studied range of pH (4.0-9.0). The results here described show an enzyme with great
stability to pH, as occurs for other cysteine proteases obtained from J. mexicana
latex, i.e. P-IV and PV (pH 3.0-10.0) (7) and that produced by C.papaya (pH 3.0-
10.0) (8).
CONCLUSIONS.
• The described strategy was appropiate to achieve the purification of P-III
protease, the third in abundance in the latex from J. mexicana.
• Econo Pac® Methyl column of hydrophobic interaction showed a greater
separation capacity compared to the strong cationic exchanger of Econo Pac ®
High S.
• The specific absorptivity by the dry weight method rendered a value of 3.2 g -1 L
cm -1 for P-III
• P-III molecular mass was 24.84 kDa determined by electrophoresis