Abstract
Background: L-asparaginase (L-AsnA) enzyme has gained significant attention in the food, biocatalysts and pharmaceutics industry. It (L-AsnA) has been widely used in food processing industries as a promising acrylamide mitigating agent and as a therapeutic agent in the treatment of certain human cancers.
Objective: Based on US Patent (4,433,054; 1984), L-asparaginase (L-AsnA) enzyme is immobilized by admixing the active enzyme on the polysaccharide to be in a gel form. The storage stability of immobilized L-AsnA enzyme and its anti-proliferation and antiviral activity were determined.
Methods: In the present study, S. maxima was cultured at large scales (300 liter) for the production of enough extracellular L-asparaginase (L-AsnA) using modified (high N concentration) Zarrouk medium as we reported in a previous study. L-AsnA was immobilized on natural polymers, as agar cake beads, agarose pieces and gelatin blocks, in order to evaluate the efficiency of physical entrapment techniques. Anti-proliferation properties of L-AsnA against lung carcinoma A549, hepatocellular carcinoma Hep-G2 and prostate carcinoma PC3 human cancer cell lines were assessed by the MTT cell viability method. In addition, the antiviral activity against Coxsackie B3 (CSB3) Virus was assessed.
Results: The highest L-AsnA immobilized activity and immobilization yield were achieved with agar cakes bead. The purified S. maxima L-AsnA showed good antiviral activity against Coxsackie B3 (CSB3) Virus in a dose-dependent manner with an IC50 value 17.03 μg/ml. The antiviral mode of action is presumably due to their capability of inhibiting attachment, blocking the adsorption and penetration event of the viral replication cycle with 89.24%, 72.78% and 72.78%, respectively. Also, S. maxima L-AsnA showed anti-proliferation effect against lung carcinoma A549, hepatocellular carcinoma Hep-G2 and prostate carcinoma PC3 human cancer cell lines, with an IC50 of 22.54, 24.65 and 56.61 μg/ml, respectively.
Conclusion: It is interesting to favor L-asparaginase of S. maxima which showed antiviral activity and anti-proliferation effect against different types of human cell lines. Thus, S. maxima microalgae might be a good source for L-AsnA enzymes and can be immobilized on natural polymers.
Keywords: L-asparaginase, anti-proliferation activity, antiviral activity, Spirulina maxima, enzyme, virus.
Graphical Abstract
[http://dx.doi.org/10.1155/2016/1785938]
[http://dx.doi.org/10.1016/j.cej.2018.08.058]
[PMID: 17285805]
[http://dx.doi.org/10.1016/j.ijbiomac.2016.11.115] [PMID: 27919811]
[http://dx.doi.org/10.25004/IJPSDR.2016.080407]
[http://dx.doi.org/10.1016/j.pep.2019.01.002] [PMID: 30615939]
[http://dx.doi.org/10.1016/j.ijbiomac.2016.07.031] [PMID: 27402458]
[http://dx.doi.org/10.1016/j.micres.2018.01.011] [PMID: 29551216]
[http://dx.doi.org/10.1016/j.ijbiomac.2018.12.162] [PMID: 30576739]
[http://dx.doi.org/10.1016/j.bcab.2013.10.005]
[http://dx.doi.org/10.1007/s13205-014-0215-z] [PMID: 28324309]
[http://dx.doi.org/10.1002/adsc.201100534]
[http://dx.doi.org/10.1021/bm400762h] [PMID: 23822160]
[http://dx.doi.org/10.1016/j.procbio.2018.09.016]
[http://dx.doi.org/10.1016/j.foodchem.2018.09.090] [PMID: 30724265]
[http://dx.doi.org/10.1080/00986445.2015.1065821]
[http://dx.doi.org/10.1007/BF02872912] [PMID: 4611872]
[http://dx.doi.org/10.4103/0250-474X.65010]] [PMID: 20838527]
[http://dx.doi.org/10.1046/j.1472-765x.1997.00331.x]
[http://dx.doi.org/10.1142/S0192415X12500607] [PMID: 22809033]
[http://dx.doi.org/10.1016/0022-1759(83)90303-4] [PMID: 6606682]
[http://dx.doi.org/10.1029/JD094iD13p16417]
[http://dx.doi.org/10.1016/j.ijbiomac.2017.11.114] [PMID: 29175524]
[http://dx.doi.org/10.1016/j.bcab.2018.11.018]
[http://dx.doi.org/10.1097/00001813-200102000-00007]] [PMID: 11261887]
[PMID: 19205431]
[http://dx.doi.org/10.1023/A:1011066129771] [PMID: 11216870]
[http://dx.doi.org/10.4236/aim.2012.22019]
[http://dx.doi.org/10.1371/journal.pone.0065250] [PMID: 23762328]
[http://dx.doi.org/10.1080/10498850.2011.654381]
[http://dx.doi.org/10.1016/j.virol.2015.06.006] [PMID: 26142496]
[http://dx.doi.org/10.1136/pgmj.49.572.428]