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首页> 外文期刊>BioTechnology: An Indian Journal >Reverse micellar extraction procedure for continuous isolation of peroxidase form plant source
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Reverse micellar extraction procedure for continuous isolation of peroxidase form plant source

机译：反胶束萃取程序可从植物来源中连续分离过氧化物酶

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Horseradish peroxidase (HRP) is the group of enzymes that catalyze the oxidation of a compound by peroxide. Horseradish peroxidase is the universal choice of reagent in molecular level experiment, diagnostics, sewage treatment and in Biosensor. Horseradish peroxidase is separated through traditional down streaming method, which usually results in low yield and high cost. Liquid-liquid extraction (LLE) using organic/aqueous phase is not the choice of separation in biotechnology mainly due solubility and protein denaturation problems.Aqueous Two Phase System (ATPS) adapts the extraction through phase separation and is considered as single step isolation protocol. TritonX-100 and Tween 20 were used as surfactants, and efficiency were calculated.Aforward extraction yield of 72%and activity 890 U/mg was obtained for Triton X-100/ Toluene at pH 6, salt concentration 0.15M, phase ratio 1:1 and surfactant concentration 15mM. In Tween 20/ toluene system there was only 58% yield in 10μl surfactant concentration. For back extraction amaximumyield of 13%and activity of 648 U/mg obtained in Triton X-100/ Toluene system.ATPSwas found to be superior to traditional methods both in terms of yield and continuous process. Isolated peroxidase was checked for its purity using SDS-PAGE, which clearly displays 3 bands correspond to the standard marker and commercial peroxidase.Gas chromatography analysis conforms the presence of peroxidase however 11 more peaks indicating presence of other proteins and enzymes. It is suggested to optimize the techniques for continues separation and concentration of the peroxidase.
机译：辣根过氧化物酶（HRP）是催化过氧化物催化化合物氧化的一组酶。辣根过氧化物酶是分子水平实验，诊断，污水处理和生物传感器中试剂的通用选择。辣根过氧化物酶通过传统的下游分离方法分离，通常导致低产量和高成本。由于溶解度和蛋白质变性问题，使用有机/水相的液-液萃取（LLE）并不是生物技术中分离的选择。水两相系统（ATPS）通过相分离适应萃取，被视为单步分离方案。以TritonX-100和Tween 20作为表面活性剂，计算了效率。在pH 6，盐浓度0.15M，相比1的条件下，Triton X-100 /甲苯的正萃取产率为72％，活性为890 U / mg。 1和表面活性剂浓度为15mM。在Tween 20 /甲苯系统中，表面活性剂浓度为10μl时，收率仅为58％。在Triton X-100 /甲苯系统中，反萃取的最大收率达13％，活性为648 U /mg。ATPSwas在产率和连续工艺方面均优于传统方法。使用SDS-PAGE检查分离的过氧化物酶的纯度，可清楚地显示3条带对应于标准标记和市售过氧化物酶。气相色谱分析表明存在过氧化物酶，但还有11个峰表明存在其他蛋白质和酶。建议优化技术以继续分离和浓缩过氧化物酶。

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