一种定量分析双歧杆菌的新方法

发布时间：2018-07-07 16:03

  本文选题：双歧杆菌 + F6PPK　； 参考：《河南农业大学》2008年硕士论文

【摘要】： 作为人和温血动物体内最重要的益生菌,双歧杆菌对宿主显示了多方面的促进健康作用。目前市场上以含有双歧杆菌的混合菌产品居多,而目前对该类产品却没有一个准确的检测方法。 本文旨在建立一种新的双歧杆菌定量分析技术。论文首先建立了双歧杆菌的培养体系,两歧双歧杆菌的培养结果显示:BBL琼脂培养基和TPY液体培养基对两歧双歧杆菌的培养效果较好,作为两歧双歧杆菌的培养基。然后进行了双歧杆菌属特征性酶F6PPK测定条件的优化。试验首先改良了原初文献(Scardovi法)F6PPK酶活测定中对照反应管的设置,又对双歧杆菌培养时间、细胞破碎方法、底物浓度、比色波长、反应温度与时间等条件进行了优化。结果表明,双歧杆菌培养18 h,收集菌体,以超声波法破碎细胞制备粗酶液,采用4 %的底物浓度,将粗酶液与底物于40℃保温30 min,最终反应物于500 nm比色,所得F6PPK酶活测定结果更可靠。 在获得优化的F6PPK酶活力测定条件的基础上,通过培养不同的时间来获取不同数量的菌体,用厌氧箱法对双歧杆菌进行平板菌落计数,用优化过的酶活测定方法测定F6PPK酶活力,分别建立了两歧双歧杆菌和长双歧杆菌的菌数与酶活力的对应关系的工作曲线。并用市场上的产品对工作曲线进行了验证,结果与平板计数法的结果一致。证明该方法是一种可靠的双歧杆菌的定量分析方法。
[Abstract]:As the most important probiotics in human and warm-blooded animals, Bifidobacterium has many health promoting effects on the host. At present, the mixed bacteria products containing Bifidobacterium are the most in the market, but there is no accurate detection method for this kind of products at present. The aim of this paper is to establish a new technique for quantitative analysis of Bifidobacterium. Firstly, the culture system of Bifidobacterium was established. The results of the culture of Bifidobacterium bifidobacterium showed that the Agar medium and TPY liquid medium had a good effect on the culture of Bifidobacterium bifidobacterium and served as the culture medium for Bifidobacterium bifidobacterium. Then, the determination conditions of characteristic enzyme F 6 PPK of Bifidobacterium were optimized. The experiment first improved the setting of control reaction tube in the determination of enzyme activity of F6PPK in original literature (Scardovi method), and optimized the conditions such as culture time of Bifidobacterium, method of cell fragmentation, substrate concentration, colorimetric wavelength, reaction temperature and time, etc. The results showed that bifidobacterium was cultured for 18 h, and the cells were broken by ultrasonic method to prepare the crude enzyme solution. The crude enzyme solution was incubated at 40 鈩，

本文编号：2105487