Antioxidants, Antimicrobial, and Anticancer Activities of Purified Chitinase of Talaromyces funiculosus Strain CBS 129594 Biosynthesized Using Crustacean Bio-Wastes

Abstract

Talaromyces funiculosus strain CBS 129594 was optimized to promote chitinase activity under solid state fermentation using crustacean bio-wastes. The aim of the study was to use purified chitinase as antioxidant, antimicrobial, and anticancer activities. The results showed that the maximum enzyme yield (2.98 ± 0.2 U/g substrate) was obtained at 1:2 crab shell chitin with the inoculation size (2.5 × 106v/v) after seven days of incubation, pH 6.5, using 0.20% of soybean meal, malt extract, and yeast extract and 100% cane and beet molasses as supplementation. The enzyme was purified with an overall yield of 7.22 purification fold with a specific activity of 9.32 ± 0.3 U/mg protein. The molecular mass of the purified chitinase was 45 kDa. The highest chitinase activity was detected at pH 6.5 and 40 °C. The purified chitinase was activated by Ca2+, Cu2+, Na+, Mn2+, and Mg2+. On the other hand, the enzyme activity was inhibited in the presence of Hg2+, Ag2+, and Li+ at 10 mM, while Zn2+ and Co2+ caused no effect compared to media without any metals. The scavenging of 2.2-diphenyl-1-picrylhydrazyl (DPPH) radicals and 2.2-pheny-l-1-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) increased with increasing the concentrations of the purified chitinase enzyme (100, 200, 300, and 400 µg/mL) which ranged from 48.7% to 57.8% and 8.87% to 63.73%, respectively. The IC50 value of DPPH radicals and ABTS of purified chitinase produced by T. funiculosus strain CBS 129594 was 199 and 306 μg/mL concentration, respectively. The purified chitinase inhibited the growth of Gram-negative bacteria (Pseudomonas aeruginosa, Escherichia coli), Gram-positive bacteria (Bacillus subtilis, Staphylococcus aureus), and fungi (Aspergillus niger, Candida albicans). The highest concentrations of purified chitinase (1000 µg/mL) caused the higher toxicity of cancer cell line MCF7 (97%), HCT116 (88.2%), and HepG2 (97.1%). In conclusion, we can conclude that chitinase can be produced from marine waste and can be used as an antioxidant, antibacterial activity, cancer therapy, and ecofriendly biocontrol agent.