溶菌酶的活性检测方法

Detection Method of Lysozyme Activity

为解决现行溶壁微球菌（比浊）法测定溶菌酶活性方法中存在的测定条件不统一、重复性不高等问题。对该方法中4个主要影响因素——溶茵酶活性范围、底物悬浮液吸光度值范围、检测温度和茵体制备方法进行了研究，以得到最佳的检测条件及菌体制备方法。结果表明：（1）溶茵酶活性75-250U／mL之间，底物悬浮液吸光度值O．45-O．77（OD450）之间，酶活力曲线具有良好的线性关系，与国家药典中基本一致；（2）检测温度25-50℃之间，底物悬浮液的吸光度变化值△OD450随温度升高而增大，适当提高检测温度可提高检测的精度，有待进一步研究；（3）用于大规模样品检测时，冷冻茵体法为制备底物悬浮液的最佳方法，检测具有良好的线性关系、管间重复性及储存稳定性。

To address the current problems of Micrococcus Lysodeikticus Fleming （turbidity） method for the determination of lysozyme activity, such as not unified measurement conditions, poor repeatability, four main influence factors had been studied which were respectively the range of lysozyme activity, substrate suspension absorbance value range, measuring temperature and cell preparation in this paper. Ultimately, the study expected to be the best test conditions and substrate preparation method. The results were as follows： （1） the lysozyme activity curves had good linear relation when the lysozyme active range of 75 U/mL to 250 U/mL and substrate suspension absorptance value range of 0.45-0.77 （wavelength 450 nm）, in general accord with the GBT 25879-2010. （2） In measuring temperature 25-50℃, substrate suspensions of absorbanee difference （/k 0945o） increased with the rise of temperature. Appropriate to improve the measuring temperature could improve the measuring accuracy, which was needed to be further studied. （3） For large-scale sample test, frozen bacteria was selected as the optimum substrate, which had good linear relation, better repeatability and storage stability.