文档介绍：论文题目:大肠杆菌内源过氧化氢检测方法的建立
学院及专业:食品与生物工程学院食科
本科生:
指导教师:
摘要
本实验旨在通过物理、化学方法的对比及探索改进,找出一种检测细菌內源过氧化氢的模型方法。
本实验以大肠杆菌突变体为研究对象,首先以30%的H2O2为研究对象,确定其原液的浓度,然后通过辣根过氧化物酶—酚红法、荧光法、硫酸钛法三种不同方法绘制出不同的标准曲线,最后根据不同的标准曲线计算出20个大肠杆菌抗氧化基因突变体菌株在同一生长时期时內源过氧化氢的释放量。
结果显示:30%,、、,根据辣根过氧化物酶—酚红法和荧光法不同的线性关系计算出来不同菌株H2O2的释放量都有明显的差异,范围在1μM~10μM之间。硫酸钛法的检测区间在10μM~500μM之间,明显检测不到大肠杆菌抗氧化基因突变体菌株中內源过氧化氢的释放量。
通过分析对比发现三种方法中辣根过氧化物酶—酚红法和荧光法都可以检测到大肠杆菌抗氧化基因突变体菌株中內源过氧化氢的释放量,硫酸钛法检测不到,但是辣根过氧化物酶—酚红法比较繁琐且需要严格控制反应条件,而荧光法的灵敏度比较高,操作也比较简单方便。
关键词:內源过氧化氢,检测方法,大肠杆菌,突变体
Subject: Measure assays establishment of endogenous hydrogen peroxide in Escherichia coli
College and Specialty: College of Food & Bioengineering
Food Science & Engineering Name:
Supervisor:
ABSTRACT
This experiment is for the purpose of discovering one detection model method to detect endogenous hydrogen peroxide by physical, chemistry method contrast and the exploration improvement.
This experiment takes the Escherichia coli mutants as the objects of study, first take 30% H2O2 as the object of study and determine the density of its original fluid, then through the horse radish peroxide enzyme - phenol red method, the fluorescence method, the titanium sulfate method three different methods draws up the different standard curve, Finally according to different standard curve to figure out endogenous hydrogen peroxide release quantity of the twenty Escherichia coli oxidation resistance gene mutant strains in the identical growth period.
The results showed that 30% H2O2 original fluid density is , use three detection methods to draw up the standard curves, and whose linear correlation coefficient respectively are , , , calculates that the H2O2 release quantity of different strains have the obvious difference according to different linear relationship of the horse radish peroxide enzyme - phenol red method and the fluorescence method, the scope is between 1μM~10μM. Th