Division of Chemistry and Biochemistry, Graduate School of Natural Science and Technology, Okayama University, 3-1-1 Tsushima-naka, Kita-ku, Okayama 700-8530, Japan.
Division of Chemistry and Biochemistry, Graduate School of Natural Science and Technology, Okayama University, 3-1-1 Tsushima-naka, Kita-ku, Okayama 700-8530, Japan.
Colloids Surf B Biointerfaces. 2017 Nov 1;159:118-124. doi: 10.1016/j.colsurfb.2017.07.074. Epub 2017 Jul 27.
Enzymatic cleaning is a potentially useful method for removing proteinaceous fouling from solid surfaces under mild conditions. Herein, the influence of an external electric field on the enzymatic cleaning of a metal surface fouled with a protein was investigated. The model fouling protein (BSA; lysozyme) was prepared on a stainless steel (St) surface, and the resulting surface subjected to enzymatic cleaning with an electric potential being applied to the St plate. Trypsin, α-chymotrypsin, and thermolysin were used as model proteases. The amounts of protein remaining on the plate before and during the cleaning process were measured by means of a reflection absorption technique using Fourier transform infrared spectroscopy. In the case for BSA fouling, the cleaning efficiency of the protease tended to increase at more negative applied potentials. Whereas, there was an optimum applied potential for removing the lysozyme fouling. Atomic force microscopy analyses indicated that applying an adequate range of electric potential enhanced the enzymatic removal of protein fouling inside scratches on the St plate surface. These findings suggest the existence of two modes of electrostatic interactions for the external electric field, one with protease molecules and the other with digested fragments of the fouling protein.
酶法清洗是一种在温和条件下从固体表面去除蛋白质污垢的潜在有效方法。本文研究了外加电场对金属表面蛋白污垢酶法清洗的影响。在不锈钢(St)表面制备模型污染蛋白(BSA;溶菌酶),然后对 St 板施加电势以进行酶法清洗。使用胰蛋白酶、α-糜蛋白酶和嗜热菌蛋白酶作为模型蛋白酶。通过使用傅里叶变换红外光谱的反射吸收技术,在清洗过程前后测量留在板上的蛋白质量。在 BSA 污染的情况下,蛋白酶的清洗效率在施加更负的电势时趋于增加。然而,去除溶菌酶污染存在最佳施加电势。原子力显微镜分析表明,施加适当范围的电势可以增强 St 板表面划痕内蛋白质污垢的酶法去除。这些发现表明外加电场存在两种静电相互作用模式,一种与蛋白酶分子有关,另一种与污染蛋白的消化片段有关。