title: Construction of Potent Recombinant Strain Through Intergeneric Protoplast Fusion in Endophytic Fungi for Anticancerous Enzymes Production Using Rice Straw.
abstract: Among all fungal endophytes isolates derived from different ethno-medical plants, the hyper-yield L-asparaginase and L-glutaminase wild strains Trichoderma sp. Gen 9 and Cladosporium sp. Gen 20 using rice straw under solid-state fermentation (SSF) were selected. The selected strains were used as parents for the intergeneric protoplast fusion program to construct recombinant strain for prompt improvement production of these enzymes in one recombinant strain. Among 21 fusants obtained, the recombinant strain AYA 20-1, with 2.11-fold and 2.58-fold increase in L-asparaginase and L-glutaminase activities more than the parental isolates Trichoderma sp. Gen 9 and Cladosporium sp. Gen 20, respectively, was achieved using rice straw under SSF. Both therapeutic enzymes L-asparaginase and L-glutaminase were purified and characterized from the culture supernatant of the recombinant AYA 20-1 strain with molecular weights of 50.6 and 83.2 kDa, respectively. Both enzymes were not metalloenzymes. Whereas thiol group blocking reagents such as p-chloromercurybenzoate and iodoacetamide totally inhibited L-asparaginase activity, which refer to sulfhydryl groups and cysteine residues involved in its catalytic activity, they have no effect toward L-glutaminase activity. Interestingly, potent anticancer, antioxidant, and antimicrobial activities were detected for both enzymes.
authors.affiliation: Department of Biological Sciences, Faculty of Sciences, King Abdulaziz University (KAU), Jeddah, 21589, Saudi Arabia.|Chemistry of Natural and Microbial Products Department, National Research Centre, Dokki, Giza, 12622, Egypt.
authors.forename: Mervat Morsy Abbas Ahmed
authors.initials: MMAA
authors.lastname: El-Gendy
authors.affiliation: Department of Biological Sciences, Faculty of Sciences, King Abdulaziz University (KAU), Jeddah, 21589, Saudi Arabia.
authors.forename: Salha Hassan Mastour
authors.initials: SHM
authors.lastname: Al-Zahrani
authors.affiliation: Genetics and Cytology Department, National Research Centre, Dokki, Giza, 12622, Egypt. ahmed_bondkly@yahoo.com.
authors.forename: Ahmed Mohamed Ahmed
authors.initials: AMA
authors.lastname: El-Bondkly
mesh_terms: D000970:Antineoplastic Agents / Q000378:metabolism*; D001215:Asparaginase / Q000096:biosynthesis / Q000235:genetics; D005972:Glutaminase / Q000096:biosynthesis / Q000235:genetics; D006801:Humans; D012275:Oryza / Q000737:chemistry*; D011523:Protoplasts / Q000201:enzymology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014242:Trichoderma / Q000201:enzymology / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D011994:Recombinant Proteins; D001215:Asparaginase; D005972:Glutaminase
keywords: Cladosporium sp.; Endophytes; Intergeneric protoplast fusion; L-Asparaginase; L-Glutaminase; Trichoderma sp.