Anticancer and antifungal efficiencies of purified chitinase produced from Trichoderma viride under submerged fermentation

• Published in: Journal of general and applied microbiology Vol. 66; no. 1; pp. 32 - 40
• Main Authors: Isaac, George Saad, Abu-Tahon, Medhat Ahmed
• Format: Journal Article
• Language: English
• Published: Japan Applied Microbiology, Molecular and Cellular Biosciences Research Foundation 01.01.2020; Japan Science and Technology Agency
• Subjects: anticancer efficiency; Antifungal activity; Biocompatibility; Body weight; Calcium; Calcium ions; Carbon dioxide; Cell lines; Chitin; Chitinase; Cobalt; Colloid chitin; Copper; Enzymes; Ethylenediaminetetraacetic acids; Fermentation; Fungi; Fungicides; Fusarium oxysporum; Lethal dose; Maltose; Mercury (metal); pH effects; Protected areas; Protein folding; Protein purification; Proteins; purification; submerged fermentation; Supplements; Tomatoes; Trichoderma viride; Wilt; Yeast; Zinc; submerged fermentation; chitinase; anticancer efficiency; antifungal activity; Trichoderma viride; purification
• ISSN: 0022-1260; 1349-8037
• DOI: 10.2323/jgam.2019.04.006

Trichoderma viride AUMC 13021 isolated from Mangrove soil of Ras Mohammed protected area at Sharm El-Sheikh, Egypt, was optimized to promote chitinase activity under submerged fermentation. The maximum enzyme yield (38.33 U/mg protein) was obtained at 1.4% of colloidal chitin, 96 h of incubation, 35°C, pH 6.5 and 125, rpm and using maltose (1%) and yeast extract (1%) as supplementation of salt basal medium. The enzyme has been purified with an overall yield of 73.1% and 5.48 purification fold, and a specific activity of 210.16 U/mg protein. The molecular mass of the purified chitinase was 62 kDa. Maximal activity of chitinase was recorded at pH 6.5 and 40°C. The highest activity was recorded in the case of colloidal chitin, with an apparent Km value of 6.66 mg/ml and Vmax of 90.8 U/ml. The purified chitinase was activated by Ca2+ and Mn2+ while the activity was inhibited by Hg2+, Zn2+, Cu2+, Co2+, dodecyl sulphate and EDTA. In vivo, the median lethal dose (LD50) was approximately 18.43 mg/kg body weight of Sprague Dawley rats. MTT assay showed that the purified chitinase has a toxic effect to MCF7 with an IC50 value 20 μg/ml, and HCT-116 cell lines with an IC50 value 44 μg/ml. Moreover, the purified enzyme showed significant antifungal activity against Fusarium oxysporum f. sp. lycopersici race 3 the causal agent of tomato wilt.