PJB-2019-453
Antifungal and anticancer efficiencies of purified chitinase produced from Trichoderma viride under submerged fermentation
Medhat Ahmed Abu-Tahon
Abstract
Trichoderma viride AUMC 13021 was optimized to enhance chitinase production under submerged culture condition. The maximum enzyme yield (38.33U/mg protein) was obtained at 1.4% of colloidal chitin, 96 h of incubation, 35oC, 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 73.1% and 5.48 purification fold and specific activity of 210.16 U/mg protein. The molecular weight of the purified chitinase was 62000 daltons, The amino acid analysis of the purified chitinase recorded eighteen amino acids. Maximal activity was recorded at pH 6.5 and 40oC. The apparent Km value was 6.66 mmol and Vmax was 90.8 Uml-1. The purified chitinase was activated by Ca+2 and Mn+2 while inhibited by Hg+2, Zn+2, Cu+2, Co+2, dodecyl sulphate and EDTA. The purified enzyme showed significant antifungal activity against Fusarium oxysporum f. sp. lycopersici race 3. Moreover, In vivo, the median lethal dose (LD50) was approximately 18.43 mg/Kg body of Sprague Dawley rats. MTT assay showed that the purified chitinase has high toxic effect to MCF7 with IC 50 value 20 μg/ml and moderate cytotoxic effect to HCT-116 cell lines with IC50 value 44 μg/ml.
To Cite this article:
Download
