O. S. Savinova, A. M. Chulkin, K. V. Moiseenko, T. V. Fedorova
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引用次数: 0
摘要
基枝菌分泌酶复合物中的过氧化物酶之所以受到关注,是因为它们具有广泛的底物特异性,而且能够参与木质素等难以降解的生物聚合物的生物降解。然而,由于很难从原生资源中分离出这些酶,因此对它们的研究十分困难。在这项工作中,创建了携带 T. hirsuta LE-BIN072 多用途过氧化物酶 VP2 编码序列的表达载体,并将其转化到 P. canescens 菌株的基因组中。对转化体的筛选表明,过氧化物酶的活性高达 1 U/mL。质谱分析鉴定了所选转化子培养液中的目标蛋白片段。首次获得了一株能产生 T. hirsuta LE-BIN072 重组多功能过氧化物酶 VP2 的新菌株 P. canescens pVP2D-6,其分泌的酶复合物具有修饰碱性木质素的能力。
Obtaining a Recombinant Producer of Trametes hirsuta Versatile Peroxidase VP2 in Penicillium canescens
The interest in peroxidases of the basidiomycete secreted enzyme complex is due to their wide substrate specificity and the ability of these enzymes to participate in the biodegradation of such difficult to degrade biopolymers as lignin. However, due to the difficulty of isolating these enzymes from native sources, their study is difficult. In this work, expression vectors were created that carried the sequence encoding the T. hirsuta LE-BIN072 versatile peroxidase VP2, which was transformed into the genome of the P. canescens strain. Screening of transformants showed the presence of peroxidase activity up to 1 U/mL. Fragments of the target protein in the culture liquids of the selected transformants were identified by mass spectrometric analysis. A new strain, P. canescens pVP2D-6, a producer of the recombinant versatile peroxidase VP2 of T. hirsuta LE-BIN072, was obtained for the first time, and the ability of the enzyme complex secreted by it to modify alkaline lignin was shown.
期刊介绍:
Applied Biochemistry and Microbiology is an international peer reviewed journal that publishes original articles on biochemistry and microbiology that have or may have practical applications. The studies include: enzymes and mechanisms of enzymatic reactions, biosynthesis of low and high molecular physiologically active compounds; the studies of their structure and properties; biogenesis and pathways of their regulation; metabolism of producers of biologically active compounds, biocatalysis in organic synthesis, applied genetics of microorganisms, applied enzymology; protein and metabolic engineering, biochemical bases of phytoimmunity, applied aspects of biochemical and immunochemical analysis; biodegradation of xenobiotics; biosensors; biomedical research (without clinical studies). Along with experimental works, the journal publishes descriptions of novel research techniques and reviews on selected topics.