PROTEASES FROM CELL CULTURE OF Bromelia hemisphaerica ...

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FSB1 – 2004 Food Science and Biotechnology in Developing Countries The specific protease activity in the callus medium (extracellular enzyme) was different for each growth-regulator combination (Figure 2). The maximal value of specific activity of callus was obtained with Picloram and BAP 0.5 mg l -1 . And these calli were selected for transfer to liquid medium to form cells suspension cultures. The results indicated that callus and biomass from cell suspension culture contain higher concentration of intracellular proteases than extracellular proteases (Figure 2). However the extracellular protease is more attractive with a view to biotechnological production. The extracellular protease activities of free cell culture medium are shown in the figure 3. The enzyme activity directly correlated with the biomass production (date not shown). In addition the extracellular protease reached the maximal value after 13 weeks decreasing subsequently to lower value (Figure 3). Proteolytic activity was showed in agar medium were calli grown (Figure 2). UT/L 18000 16000 14000 12000 10000 8000 6000 4000 2000 0 MS4 MS5 MS8 MS12 CALLUS Murashige & Skoog Medium Figure 2. Extracellular and intracellular (calli in MS12) protease activity of calli of B. hemisphaerica grown in different growth regulators proportion, Auxins 0.5 mg l -1 . Activity expressed in terms of tyrosine (UT/L). Cell suspension culture medium had extacellular proteolytic activity of 2000 – 20000 UT/L (Figure 3).
UT/L 25000 20000 15000 10000 5000 FSB1 – 2004 Food Science and Biotechnology in Developing Countries . 0 0 2 4 6 8 10 12 Weeks 14 16 18 20 22 24 Figure 3. Extracellular protease of cell suspension cultures of B. hemisphaerica. MS 12: Picloram and BAP 0.5 mg l -1 . Activity expressed in terms of tyrosine (UT/L). CONCLUSION In our opinion callus and cell suspension cultures of B. hemisphaerica show promising potential for the production of cysteine protease that might be suitable for the transformation of foods. Acknowledgements This work was supported by Mexican National Polytechnic Institute (IPN) Project CGPI: 20030510. The authors would like to thank: 1a,2a SIBE-COFFA-IPN, 1b PIFI-IPN,
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PROTEASES FROM CELL CULTURE OF Bromelia hemisphaerica ...

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