Department of Agriculture and Food Systems, Melbourne School of Land and Environment, University of Melbourne, Parkville, VIC 3010, Australia.
J Agric Food Chem. 2010 Nov 24;58(22):11725-31. doi: 10.1021/jf102171u. Epub 2010 Oct 19.
Pulsed electric field (PEF) treatment (35 kV cm(-1) for 19.2 μs using bipolar 2 μs pulses) was conducted on bovine lactoferrin (LF; 0.4 mg mL(-1)) prepared in simulated milk ultrafiltrate (SMUF), at concentrations between 0.2× and 2× normal strength, with electrical conductivities ranging from 0.17 to 1.04 S m(-1). The physicochemical and structural characteristics (LF content by a spectrophotometric and an ELISA method, surface hydrophobicity, electrophoretic mobility, far-UV circular dichroism spectra, and tryptophan fluorescence) of LF dissolved in SMUF of all strengths tested were not changed after PEF treatment. The PEF treatment of LF in 0.2 strength SMUF did not cause the release of LF-bound ferric ion into the aqueous phase, with a concentration of LF-bound iron being the same as that of the untreated LF control (174 μg L(-1)). However, in treatment media with higher ionic strengths, ferric ion was released from the LF molecule into the aqueous phase. The concentration of LF-bound iron decreased from 174 μg L(-1) for the LF treated in 0.2 strength SMUF to 80 μg L(-1) for that treated in double-strength SMUF. The results suggest that the PEF-induced iron depletion of LF does not appear to cause an appreciable conformational change in LF molecules. PEF treatment could be developed as a novel physical way to produce iron-depleted LF, as an alternative to the existing chemical method.
脉冲电场(PEF)处理(使用双极 2 μs 脉冲,35 kV cm(-1) 处理 19.2 μs)在模拟乳超滤物(SMUF)中进行,浓度范围为 0.2×至 2×正常强度,电导率范围为 0.17 至 1.04 S m(-1)。在所有测试强度的 SMUF 中,LF 的物理化学和结构特性(分光光度法和 ELISA 法测定 LF 含量、表面疏水性、电泳迁移率、远紫外圆二色光谱和色氨酸荧光)在 PEF 处理后没有变化。在 0.2 强度 SMUF 中 PEF 处理 LF 不会导致 LF 结合的铁离子释放到水相中,LF 结合铁的浓度与未处理 LF 对照(174 μg L(-1))相同。然而,在离子强度较高的处理介质中,铁离子从 LF 分子中释放到水相中。LF 结合铁的浓度从 0.2 强度 SMUF 中处理的 LF 从 174 μg L(-1) 降低到双强度 SMUF 中处理的 LF 从 80 μg L(-1)。结果表明,PEF 诱导 LF 中铁的耗尽似乎不会引起 LF 分子的明显构象变化。PEF 处理可以作为一种新的物理方法来生产缺铁 LF,作为现有化学方法的替代方法。