Spirulina maxima L-asparaginase: Immobilization, Antiviral and Antiproliferation Activities

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.

Recent Strategies and Applications for l-Asparaginase Confinement

“…Despite becoming space-restricted, the enzymes remain free in movement, while small substrates can freely cross the semi-permeable membrane [132,133]. Baky and Baroty [87] successfully entrapped ASNase from Spirulina maxima onto natural polymers, within which agar cake beads displayed the highest ASNase ηi and confined enzymatic activity [87].…”

“…Baky and Baroty [87] successfully entrapped ASNase from Spirulina maxima onto natural polymers, within which agar cake beads displayed the highest ASNase η i and confined enzymatic activity [87].…”

“…Recent in vitro screening of confined ASNase in distinct cancer cell lines, namely lung cancer cell line (A549) using AuNPs [76], cerium selenium nanobiocomposite [84] and natural polymers such as agar cake beads [87]; ovarian cancer cell line (A2780) via AuNPs [76]; brain cancer cell lines (U87) and cervical cancer cell lines (HeLa) through β-cyclodextrin-gelatin nanobiocomposite [86]; prostate cancer cell lines (PC3) applying β-cyclodextrin nanobiocomposite [85] and natural polymers [87]; human myeloid leukaemia cell line (U937) with β-cyclodextrin nanobiocomposite [85]; ALL cell line (MOLT-4) using AONP [77]; and hepatocellular carcinoma (Hep-G2) cancer cell lines using natural polymers [87] displayed promising anticancer activity (detailed in Table 4). Regarding ASNase from A. terreus, while the highest in vitro cytotoxicity levels (84.51%) were obtained with AuNPs against A549 cell line [77], the in vitro cytotoxicity levels of AuNPs and free ASNase against the same cell line were already high (73.68% and 74.88%, respectively), thus the in vitro cytotoxicity levels only increased by 10.83% and 9.63%, respectively.…”

l-asparaginase: Need for an Expedition from an Enzymatic Molecule to Antimicrobial Drug

“…N-Bromosuccinimide-modified l -asparaginase has shown greater stability and prolonged half-life (144 h) in vitro to proteolytic digestion as compared to unmodified l -asparaginase enzyme (93 h) (Mohan Kumar et al 2014 ). Recently antiviral activity of this enzyme was shown against Coxsackie B3 Virus (Abd El-Baky and El-Baroty et al 2020 ). They have advocated the antiviral capability of l -asparaginase via inhibiting attachment, blocking the adsorption, and penetrating viral replication.…”

“…(Vimal and Kumar 2017 ). We have noticed an interesting fact from the literature that l -asparaginase was explored as an anti-infectious agent also (Vimal and Kumar 2021 ; Abd El-Baky and El-Baroty 2020 ; Meganathan 2016 ). Therefore, we have tried to provide an outlook for the further development of biopolymer-based l -asparaginase nanoformulation as an emerging and alternative therapy against microbial infections.…”