Santucci Roberto, Laurenti Enzo, Sinibaldi Federica, Ferrari Rosa Pia
Dipartimento di Medicina Sperimentale e Scienze Biochimiche, Università di Roma 'Tor Vergata', Via Montpellier 1, 00133, Rome, Italy.
Biochim Biophys Acta. 2002 Apr 29;1596(2):225-33. doi: 10.1016/s0167-4838(02)00220-0.
Electrochemical biosensors have found wide application in food and clinical areas, as well as in environmental field. A large number of articles focused on horseradish peroxidase (HRP)-based biosensors have been published in the last decade, due to the capability of HRP to quantitatively detect the presence of hydrogen peroxide produced in a reaction. At present a large body of multi-enzymatic amperometric biosensors are realized by entrapping HRP together with other enzymes into a polymeric matrix; these systems represent a promising example of simple, low-cost electrochemical tools for the analysis of bioanalytes in solution, such as glucose, choline and cholesterol. Due to the fact that polymers used for HRP entrapping are soluble in organic solvents and that many solvents are strong denaturants of aquo-soluble proteins, in this paper we investigate (in particular, by circular dichroism and electron paramagnetic spectroscopies) the effect of dimethyl sulfoxide, one of the organic solvents employed for polymer solubilization, on the structure and the functionality of HRP, in order to determine the effect induced by the solvent concentration on the structure and activity of the hemoprotein. This is relevant for basic and applied biochemistry, HRP being largely employed in bioinorganic chemistry and sensor area.
电化学生物传感器已在食品、临床领域以及环境领域得到广泛应用。在过去十年中，由于辣根过氧化物酶（HRP）能够定量检测反应中产生的过氧化氢的存在，大量聚焦于基于HRP的生物传感器的文章得以发表。目前，通过将HRP与其他酶一起包埋在聚合物基质中，实现了大量的多酶安培生物传感器；这些系统是用于分析溶液中生物分析物（如葡萄糖、胆碱和胆固醇）的简单、低成本电化学工具的一个有前景的例子。由于用于包埋HRP的聚合物可溶于有机溶剂，且许多溶剂是水溶性蛋白质的强变性剂，在本文中我们（特别是通过圆二色性和电子顺磁共振光谱法）研究了用于聚合物增溶的有机溶剂之一二甲基亚砜对HRP结构和功能的影响，以确定溶剂浓度对血蛋白结构和活性的影响。这对于基础生物化学和应用生物化学而言至关重要，因为HRP在生物无机化学和传感器领域大量应用。
