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Pectin Methylesterase of Datura species purification and characterization from Datura stramoniumand its application

机译:曼陀罗属果胶的果胶甲基酯酶的纯化鉴定及其应用

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摘要

Pectin methylesterases (PME; EC 3.1.1.11) involved in de-esterification of pectin and have applicability in food, textiles, wines, pulp, and paper industries. In the present study, we compared PME activity of different parts of 3 Datura species and found that fruit coat showed maximum PME activity followed by leaf and seed. PME from leaves of D. stramonium (DsPME) was purified and characterized. DsPME showed optimum activity at 60 °C and pH 9 in the presence of 0.3 M NaCl. DsPME was stable at 70 °C and retained more than 40% activity after 60 min of incubation. However, enzyme activity completely abolished at 80 after 5 min of incubation. It follows Michaelis-Menten enzyme kinetics. Km and Vmax with citrus pectin were 0.008 mg/ml and 16.96 µmol/min, respectively. DsPME in combination with polygalactourenase (PGA) increased the clarity of orange, apple, pomegranate and pineapple juices by 2.9, 2.6, 2.3, and 3.6 fold, respectively in comparison to PGA alone. Due to very high de-esterification activity, easy denaturation and significant efficacy in incrementing clarification of fruit juice makes DsPME useful for industrial application.
机译:果胶甲基酯酶(PME; EC 3.1.1.11)参与果胶的去酯化反应,可用于食品,纺织品,葡萄酒,纸浆和造纸工业。在本研究中,我们比较了3种曼陀罗物种不同部位的PME活性,发现果皮表现出最大的PME活性,其次是叶片和种子。纯化并鉴定了来自D. stramonium(DsPME)叶片的PME。在存在0.3 M NaCl的情况下,DsPME在60°C和pH 9时显示最佳活性。 DsPME在70°C下稳定,孵育60分钟后保留超过40%的活性。但是,孵育5分钟后,酶活性在80时完全消失。它遵循Michaelis-Menten酶动力学。柑橘果胶的Km和Vmax分别为0.008 mg / ml和16.96 µmol / min。与单独的PGA相比,DsPME与聚半乳糖醛酸酶(PGA)的结合使橙汁,苹果汁,石榴汁和菠萝汁的澄清度分别增加了2.9倍,2.6倍,2.3倍和3.6倍。由于很高的去酯化活性,易于变性和在提高果汁澄清度方面的显着功效,使得DsPME可用于工业应用。

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