本文關(guān)鍵詞：抑制引起腹瀉致病菌的益生菌篩選及機(jī)制初步研究　出處：《哈爾濱工業(yè)大學(xué)》2010年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 益生菌篩選 蛋白質(zhì)純化 抑制黏附 致病性大腸桿菌

【摘要】： 腹瀉是發(fā)展中國(guó)家導(dǎo)致兒童死亡的主要原因,每年有大約140萬(wàn)-250萬(wàn)的生命死于腹瀉。腹瀉除了直接引起健康的損傷外,還引起人體營(yíng)養(yǎng)失調(diào),生長(zhǎng)速度減緩,智力發(fā)育等問(wèn)題。盡管在發(fā)達(dá)國(guó)家腹瀉導(dǎo)致死亡的的情況并不經(jīng)常發(fā)生,但是腹瀉仍然是引起普通疾病的主要原因,而且還消耗了大量的醫(yī)療費(fèi)用。我們篩選能夠抑制引起腹瀉致病菌的益生菌,這些益生菌對(duì)于預(yù)防腹瀉發(fā)揮重要的作用,益生菌配合藥物治療腹瀉既能夠使腹瀉快速康復(fù),又能夠保護(hù)腸道菌群免受藥物的破壞。用益生菌預(yù)防和治療腹瀉還能夠有效地降低醫(yī)療費(fèi)用,減輕醫(yī)療負(fù)擔(dān)。 從健康成人腸道中分離腸道益生菌。采集糞便樣本,稀釋之后用RB、CAN和LAM培養(yǎng)基分離出39株益生菌,其中15株雙歧桿菌、14株乳酸球菌和10株乳酸桿菌。然后以致病性大腸桿菌(E.coli)作為指標(biāo)菌株對(duì)這39株益生菌進(jìn)行抑菌篩選,發(fā)現(xiàn)乳酸球菌對(duì)于致病性大腸桿菌沒(méi)有抑制作用,大部分雙歧桿菌和乳桿菌有抑菌作用。對(duì)于剩余的具有抑菌能力的益生菌菌液上清進(jìn)行中和,發(fā)現(xiàn)大部分益生菌喪失抑菌能力,僅有三株益生菌具有較強(qiáng)的抑菌活性,最后通過(guò)過(guò)氧化氫酶試驗(yàn)和蛋白酶檢測(cè)試驗(yàn)確定1321、0533和1333這三株菌,不僅能夠代謝產(chǎn)生各種酸抑菌,還能夠產(chǎn)生抗菌肽抑制致病菌的生長(zhǎng)。最后通過(guò)蛋白質(zhì)純化儀對(duì)于益生菌代謝產(chǎn)物中的抗菌肽進(jìn)行分離純化。 篩選出能夠抑制致病菌的益生菌后還要對(duì)益生菌的耐酸、耐膽鹽和黏附能力進(jìn)行篩選,因?yàn)橹挥幸嫔軌蚰褪苋梭w胃和腸道的復(fù)雜環(huán)境,順利黏附到人體小腸上皮細(xì)胞上才能發(fā)揮作用。最后對(duì)于益生菌抑制致病菌黏附進(jìn)行了初步研究,驗(yàn)證了益生菌通過(guò)代謝產(chǎn)物抑制黏附抑制腹瀉的機(jī)制。
[Abstract]:Diarrhoea is the leading cause of death among children in developing countries, with about 1.4 million-2.5 million deaths per year due to diarrhoea, which, in addition to direct health damage, causes nutritional disorders in the human body. Problems such as slower growth and mental development. Although diarrhoea is not a common cause of death in developed countries, diarrhoea is still the leading cause of common diseases. We screen probiotics that inhibit the bacteria that cause diarrhoea, which play an important role in preventing diarrhea. Probiotics combined with drugs to treat diarrhea can not only make rapid recovery of diarrhea, but also protect intestinal flora from drug damage. Probiotics to prevent and treat diarrhea can also effectively reduce the cost of medical treatment. Lighten the burden of medical care. Intestinal probiotics were isolated from the intestinal tract of healthy adults. Fecal samples were collected, and 39 strains of probiotics were isolated by dilution with RBCAN and LAM medium, of which 15 strains were bifidobacterium. 14 strains of Lactococcus lactis and 10 strains of Lactobacillus were used as indicator strains of E. coli to screen 39 strains of probiotics. It was found that Lactococcus had no inhibitory effect on pathogenic Escherichia coli and most of Bifidobacterium and Lactobacillus had bacteriostasis. The supernatant of the remaining probiotic bacteria with bacteriostasis was neutralized. It was found that most probiotics lost their bacteriostatic ability, and only three probiotics had strong bacteriostatic activity. Finally, 1321 was determined by catalase test and protease test. The three strains, 0533 and 1333, can not only metabolize and produce a variety of acid antibacterial bacteria. Antimicrobial peptides can also be produced to inhibit the growth of pathogenic bacteria. Finally, the antimicrobial peptides in the metabolites of probiotics were separated and purified by protein purification apparatus. After screening probiotics which can inhibit pathogenic bacteria, we have to screen the acid resistance, bile salt tolerance and adhesion ability of probiotics, because only probiotics can tolerate the complex environment of human stomach and intestine. Finally, a preliminary study on the inhibition of bacterial adhesion by probiotics was carried out to verify the mechanism of the inhibition of diarrhea by metabolites of probiotics.
【學(xué)位授予單位】：哈爾濱工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類(lèi)號(hào)】：R371

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中國(guó)期刊全文數(shù)據(jù)庫(kù) 前10條

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