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Obtaining mutant fungal strains of Aspergillus niger with high production of fructooligosaccharides (FOS) using ultraviolet light irradiation

机译:使用紫外线照射获得高产低聚果糖(FOS)的黑曲霉突变菌菌株

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A strain of Aspergillus niger PR-142 native to northern coast of Peru was subjected to successive processes of mutagenesis by ultraviolet light (UV) irradiation at 253.7 nm to increase the production of fructooligosaccharides (FOS). An initial selection was made by considering the mutants with increased invertase activity followed by the measurement of β-fructosyltransferase (FTase) activity both in mycelium and extracellular environment. Five selected mutants, which showed increased values of mycelium invertase activity (ranging from 101 to 128% as compared to the parent strain) at 40°C and sodium dodecylsulfate 0.15 (w/v), were? grown in a fermentative medium in 50 mL conical tubes on a rotary shaker, and their FTase activity was determined. The 6-M69 mutant showed the most active mycelium activity of 1.5 fold as compared to the parent strain. When the same reaction was performed between 1 to 4 h, at the 3rd h, the mycelium FTase activity significantly increased up to 7 and 3 times in the mutant and parental strain, respectively. Finally, 4 mutants and the parental PR-142 were genetically characterized using inter simple sequence repeat polymerase chain reaction (ISSR-PCR) molecular markers. This analysis showed a significant 33% polymorphic bands between the parent and mutant markers, and 20 bands were unique to the mutants.
机译:秘鲁北部海岸的黑曲霉PR-142菌株通过253.7 nm的紫外线(UV)辐照进行了连续诱变过程,以增加低聚果糖(FOS)的产量。通过考虑具有增加的转化酶活性的突变体,然后在菌丝体和细胞外环境中测量β-果糖基转移酶(FTase)活性来进行初步选择。选择的五个突变体在40°C和十二烷基硫酸钠0.15(w / v)下显示的菌丝体转化酶活性值增加(与亲本菌株相比,从101%到128%)。在旋转摇床上在50 mL锥形管中的发酵培养基中培养,测定其FTase活性。与亲本菌株相比,6-M69突变体显示出最活跃的菌丝体活性1.5倍。当相同的反应在1至4小时之间进行时,在第3小时,突变体和亲本菌株的菌丝体FTase活性分别显着提高了7倍和3倍。最后,使用简单序列重复聚合酶链反应(ISSR-PCR)分子标记对4个突变体和亲代PR-142进行了遗传鉴定。该分析表明,亲本和突变体标记之间存在显着的33%多态性条带,而突变体独有20条条带。

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