本文選題：青春雙歧桿菌 + 耐氧馴化��； 參考：《東北林業(yè)大學(xué)》2010年碩士論文

【摘要】： 本研究目的是從嬰兒糞便中分離出青春雙歧桿菌,經(jīng)過(guò)生化及分子鑒定后,對(duì)此分離菌株進(jìn)行抗逆環(huán)境馴化及篩選,對(duì)篩選出的優(yōu)良菌株進(jìn)行培養(yǎng)條件的優(yōu)化,從而滿足工業(yè)化生產(chǎn)的技術(shù)需求。為了確保其安全性,對(duì)該菌株進(jìn)行毒理試驗(yàn),同時(shí)進(jìn)行功能性測(cè)試,并開發(fā)益生菌相關(guān)應(yīng)用產(chǎn)品。 分離得到的疑似目的菌株,通過(guò)特征酶及發(fā)酵糖試驗(yàn)進(jìn)行菌種屬的篩選,再進(jìn)一步應(yīng)用分子生物學(xué)方法,提取菌株16S rDNA,通過(guò)基因序列的比對(duì),對(duì)菌株進(jìn)行種的鑒定。成功分離出目的菌株——青春雙歧桿菌,通過(guò)發(fā)育樹系統(tǒng)命名為：Bifidobacterium adolescentis BA005。 通過(guò)無(wú)氧-有氧交替馴化的方法,篩選出耐氧能力較強(qiáng)的青春雙歧桿菌菌株,馴化后菌株的菌落形態(tài)和顯微鏡觀察狀態(tài)基本沒(méi)有改變,菌體生長(zhǎng)速度較無(wú)氧培養(yǎng)時(shí)緩慢,生長(zhǎng)曲線中對(duì)數(shù)生長(zhǎng)期拖后,但對(duì)培養(yǎng)后的活菌數(shù)影響不大,有氧培養(yǎng)24h后活菌數(shù)可達(dá)到3.30×1010 cfu/mL。 為降低工業(yè)生產(chǎn)中的生產(chǎn)成本,本研究采用玉米汁為培養(yǎng)基質(zhì),對(duì)耐氧Bifidobacterium adolescentis BA005進(jìn)行培養(yǎng)條件響應(yīng)面法優(yōu)化。確定培養(yǎng)的最佳條件：接種量5%,玉米糖化液濃度12.1Bx,初始pH值6.48,培養(yǎng)溫度37℃,發(fā)酵時(shí)間20h。在此條件下菌液最大OD值為1.819,活菌數(shù)為3.05×1012 cfu/mL。 人工模擬人胃、腸道酸性及酶環(huán)境,對(duì)耐氧Bifidobacterium adolescentis BA005進(jìn)行抗胃腸道逆環(huán)境菌株篩選,并對(duì)常用抗生素耐性進(jìn)行測(cè)試。結(jié)果表明,在模擬胃液環(huán)境中菌存活率為72.12%；在模擬腸道環(huán)境中菌存活率為53.33%；對(duì)大多數(shù)常用抗生素僅為中度敏感,對(duì)氨芐西林具有較強(qiáng)的耐藥性。證明此菌株可以發(fā)揮其保健作用,維持人體健康。 對(duì)Bifidobacterium adolescentis BA005進(jìn)行小鼠急性毒理試驗(yàn)及短期(30天)喂養(yǎng)試驗(yàn),試驗(yàn)劑量超過(guò)了人可能食用劑量的10倍,試驗(yàn)小鼠沒(méi)有出現(xiàn)中毒或死亡等異常狀況,解剖其內(nèi)臟主要器官,亦未見異常。從而證明此菌為安全菌株,可以放心食用并進(jìn)行開發(fā)利用。 通過(guò)藥物建造腹瀉和便秘小鼠模型,對(duì)模型小鼠給予不同種類的益生菌劑,通過(guò)比較,Bifidobacterium adolescentis BA005菌株對(duì)腹瀉和便秘癥狀的緩解和治療功效最強(qiáng)。 基于青春雙歧桿菌的諸多功能性,對(duì)其進(jìn)行新產(chǎn)品開發(fā)。產(chǎn)品之一為格瓦斯發(fā)酵飲料,通過(guò)雙歧桿菌和酵母菌的共同發(fā)酵,飲料中含有豐富的營(yíng)養(yǎng)物質(zhì),酸甜適口；產(chǎn)品之二為功能性發(fā)酵飼料,通過(guò)雙歧桿菌和酵母菌共同對(duì)常規(guī)飼料進(jìn)行發(fā)酵,產(chǎn)生更利于動(dòng)物吸收利用的營(yíng)養(yǎng)物質(zhì)。喂養(yǎng)奶�？梢蕴岣咂鋵�(duì)常規(guī)飼料的消化吸收率,平衡胃腸道微生態(tài)系統(tǒng),增強(qiáng)免疫力。
[Abstract]:The purpose of this study was to isolate Bifidobacterium pubertifolia from infant feces. After biochemical and molecular identification, the strains were acclimated and screened in the stress resistant environment, and the culture conditions of the excellent strains were optimized. In order to meet the technical needs of industrial production. In order to ensure its safety, toxicological tests, functional tests and probiotic application products were carried out. Strain 16s rDNA was extracted by molecular biology method, and the strain was identified by comparison of gene sequence. The strain was screened by characteristic enzyme and fermentation sugar test, and the strain 16s rDNA was extracted by molecular biology method. Bifidobacterium adolescentis BA005. The strain of Bifidobacterium youth with strong oxygen tolerance was screened by the method of anaerobic and aerobic alternate acclimation. After acclimation, the colony morphology and microscopic observation state of the acclimated strain were basically unchanged, and the growth rate of the strain was slower than that of the anaerobic culture. The logarithmic growth period in the growth curve was delayed, but it had little effect on the number of viable bacteria. After 24 hours of aerobic culture, the number of viable bacteria could reach 3.30 脳 1010 cfu-mL. In order to reduce the production cost in industrial production, the culture condition response surface method (RSM) was used to optimize the oxygen tolerant Bifidobacterium adolescentis BA005 culture by using corn juice as the culture substrate. The optimum culture conditions were determined as follows: inoculation amount 5, corn saccharification solution concentration 12.1Bx, initial pH 6.48, culture temperature 37 鈩，

本文編號(hào)：1904967