本文選題：雙歧桿菌 + F6PPK��； 參考：《河南農(nóng)業(yè)大學(xué)》2008年碩士論文

【摘要】： 作為人和溫血?jiǎng)游矬w內(nèi)最重要的益生菌,雙歧桿菌對(duì)宿主顯示了多方面的促進(jìn)健康作用。目前市場(chǎng)上以含有雙歧桿菌的混合菌產(chǎn)品居多,而目前對(duì)該類(lèi)產(chǎn)品卻沒(méi)有一個(gè)準(zhǔn)確的檢測(cè)方法。 本文旨在建立一種新的雙歧桿菌定量分析技術(shù)。論文首先建立了雙歧桿菌的培養(yǎng)體系,兩歧雙歧桿菌的培養(yǎng)結(jié)果顯示:BBL瓊脂培養(yǎng)基和TPY液體培養(yǎng)基對(duì)兩歧雙歧桿菌的培養(yǎng)效果較好,作為兩歧雙歧桿菌的培養(yǎng)基。然后進(jìn)行了雙歧桿菌屬特征性酶F6PPK測(cè)定條件的優(yōu)化。試驗(yàn)首先改良了原初文獻(xiàn)(Scardovi法)F6PPK酶活測(cè)定中對(duì)照反應(yīng)管的設(shè)置,又對(duì)雙歧桿菌培養(yǎng)時(shí)間、細(xì)胞破碎方法、底物濃度、比色波長(zhǎng)、反應(yīng)溫度與時(shí)間等條件進(jìn)行了優(yōu)化。結(jié)果表明,雙歧桿菌培養(yǎng)18 h,收集菌體,以超聲波法破碎細(xì)胞制備粗酶液,采用4 %的底物濃度,將粗酶液與底物于40℃保溫30 min,最終反應(yīng)物于500 nm比色,所得F6PPK酶活測(cè)定結(jié)果更可靠。 在獲得優(yōu)化的F6PPK酶活力測(cè)定條件的基礎(chǔ)上,通過(guò)培養(yǎng)不同的時(shí)間來(lái)獲取不同數(shù)量的菌體,用厭氧箱法對(duì)雙歧桿菌進(jìn)行平板菌落計(jì)數(shù),用優(yōu)化過(guò)的酶活測(cè)定方法測(cè)定F6PPK酶活力,分別建立了兩歧雙歧桿菌和長(zhǎng)雙歧桿菌的菌數(shù)與酶活力的對(duì)應(yīng)關(guān)系的工作曲線(xiàn)。并用市場(chǎng)上的產(chǎn)品對(duì)工作曲線(xiàn)進(jìn)行了驗(yàn)證,結(jié)果與平板計(jì)數(shù)法的結(jié)果一致。證明該方法是一種可靠的雙歧桿菌的定量分析方法。
[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，

本文編號(hào)：2105487