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首页> 外文期刊>Applied biochemistry and biotechnology, Part A. enzyme engineering and biotechnology >Construction of Potent Recombinant Strain Through Intergeneric Protoplast Fusion in Endophytic Fungi for Anticancerous Enzymes Production Using Rice Straw
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Construction of Potent Recombinant Strain Through Intergeneric Protoplast Fusion in Endophytic Fungi for Anticancerous Enzymes Production Using Rice Straw

机译：用稻草抗生素真菌抗生素真菌融合效应效应效应

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摘要

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.
机译：在所有真菌内心细胞中，来自不同的民族医疗植物，超产量L-天冬酰胺酶和L-谷氨酰胺酶野生菌株Trichoderma sp。 11型和囊孢子sp。选择在固态发酵（SSF）下使用稻草的Gen 20。所选菌株用作母体用于代苯基原生质体融合程序，以构建重组菌株以提高一种重组菌株在一种重组菌株中提高这些酶的产生。在获得的21个杂散中，重组菌株Aya 20-1，L-天冬酰胺酶和L-谷氨酰胺酶活性的2.11倍和2.58倍的增加，比亲本分离株trichoderma sp更多。 11型和囊孢子sp。分别在SSF下使用稻草获得Gen 20。纯化治疗性酶L-天冬酰胺酶和L-谷氨酰胺酶，分别从重组AYA 20-1株的培养上清液中纯化，分别为50.6和83.2kDa的分子量。两种酶都不是金属酶。虽然硫醇基团被阻断试剂如p-氯酸盐脱苯甲酸酯和碘乙酰胺完全抑制L-天冬酰胺酶活性，其参考其催化活性涉及巯基和半胱氨酸残基，它们对L-谷氨酰胺酶活性没有影响。有趣的是，为两种酶检测有效的抗癌，抗氧化剂和抗微生物活性。

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