Biochemical characterization and antitumor activity of three phase partitioned l-asparaginase from Capsicum annuum L

• Published in: Separation and purification technology 2015-03, Vol.142, p.258-267
• Main Authors: Shanmugaprakash, M., Jayashree, C., Vinothkumar, V., Senthilkumar, S.N.S., Siddiqui, S., Rawat, V., Arshad, M.
• Format: Article
• Language: English
• Subjects: Anticancer properties; Antiproliferative activity; Antiproliferatives; Capsicum annuum; Enzymes; Flow-cytometry; Homogeneity; l-asparaginase; Partitioning; Purification; Three phase; Three phase partitioning
• ISSN: 1383-5866; 1873-3794
• DOI: 10.1016/j.seppur.2014.12.036

[Display omitted] •A rapid and single step process was developed for purification of l-asparaginase.•The optimum temperature and pH of enzyme was determined as 40°C and 8.5.•The enzyme showed good scavenging activity and the total antioxidant capacity.•The purified TPP l-asparaginase showed anti-proliferative activity in cancer cell lines.•The IC50 for (Hela, A549, and KB) were evaluated. In this study, three phase partitioning (TPP) as an efficient bioseparation method, was explored for the first time to purify stable uncontaminated l-asparaginase from Capsicum annuum L., and its antitumor activity was evaluated. With the optimized system parameters, the enzyme was purified to homogeneity and having 6.83-fold with 567.4% recovery of its activity. The optimum pH and temperature of the TPP purified enzyme were determined as 8.5 and 40°C, respectively. The stability studies of the enzyme activity envisaged that the enzyme is stable up to 45°C and retained its activity over a wide range of pH (5.0–9.0). The purified enzyme showed good scavenging activity, and its total antioxidant capacity was found to be 88.58μM ascorbic acid equivalent. The antiproliferative activity of the purified l-asparaginase was also investigated against the three human cancerous cell lines.