The presented study aims to the screening of the endophytic fungi associated with Saint Katherine plants to produce L-asparaginase enzyme. Twenty-five endophytic fungal species were isolated and identified from a twenty-three-plant species using surface sterilization techniques. Plant species were collected from nine wadis over Saint Katherine protectorate (SKP), during the period of 2015-2017` with a total count of 4466 CFU. Five species with nine morphotypes showed L-asparaginase activity ranging from 44.5 (±1.66) to 152.58 (±0.63) U/ml. Lasiodiplodia theobromae recoded the highest activity with 152.58 U/ml. L-asparaginase was induced with an activity of 315 U/ml. L -asparaginase was purified with a specific activity of 468.03 U/mg and a total activity of 84.4 U/ml. Purified L-asparaginase showed an approximate size of 70 kDa. Purified enzyme was characterized chemically. Purified L-asparaginase showed an IC50 of 35.2±0.7 U/ml with leukemic M-NFS-60 cell lines and CC50 of 79.4±1.9 U/ml with normal WI-38 cell line. The presented study suggests the use of endophytic fungi as a sustainable source for L-asparaginase production.
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