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Effects of Pulsed Electric Fields on the Activity and Structure of Pepsin.

机译:脉冲电场对胃蛋白酶活性和结构的影响。

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A continuous pulsed electric field (PEF) system integrated with six co-field flow PEF treatment chambers was used to study the inactivation of pepsin. The inactivation of pepsin activity was a function of applied electric field strength, electrical conductivity, and pH. The inactivation of pepsin by PEF followed a first-order model. The first-order inactivation kinetic constant of pepsin was 0.012 (1/mus) in 7.5 mM HCl (pH 2.0) at 34.2 kV/cm. Aggregation of pepsin was observed during PEF treatment; however, the inactivation took place before the formation of aggregates. Circular dichroism analysis showed that inactivation of pepsin by PEF was correlated to the loss of beta-sheet structure in a pepsin molecule. The relative residual activity of PEF-treated pepsin was correlated to the relative molar ellipticity at 215 nm. Both PEF- and heat-induced inactivation of pepsin were correlated with the alteration of the secondary structure (beta-sheet dominant structure) of pepsin.
机译:连续脉冲电场(PEF)系统与六个同场流动PEF处理室集成在一起,用于研究胃蛋白酶的灭活。胃蛋白酶活性的失活是施加的电场强度,电导率和pH的函数。 PEF对胃蛋白酶的灭活遵循一阶模型。在34.2 kV / cm的7.5 mM HCl(pH 2.0)中,胃蛋白酶的一级失活动力学常数为0.012(1 / mus)。在PEF治疗期间观察到胃蛋白酶的聚集;然而,灭活发生在聚集体形成之前。圆二色性分析表明,PEF使胃蛋白酶失活与胃蛋白酶分子中β-折叠结构的丧失有关。 PEF处理的胃蛋白酶的相对残留活性与215 nm处的相对摩尔椭圆率相关。 PEF和热诱导的胃蛋白酶失活均与胃蛋白酶的二级结构(β-表位显性结构)的改变有关。

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