本文選題：麩皮多糖 + 微生物發(fā)酵��； 參考：《內(nèi)蒙古農(nóng)業(yè)大學(xué)》2017年碩士論文

【摘要】：本研究以麩皮為原料,配以豆粕粉、玉米粉等輔料,利用微生物發(fā)酵法制備麩皮多糖并對其粗制品抗氧化活性進行研究。本論文從以下兩個方面進行研究探討:1麩皮多糖的微生物發(fā)酵制備和提取1.1發(fā)酵菌種及其組合的篩選以發(fā)酵麩皮中多糖產(chǎn)量為指標,利用植物乳桿菌P8(LactobacillusplantarumP8)、釀酒酵母 CGMCC2.119(Saccharomyces cerevisiae CGMCC2.119)、枯草芽孢桿菌CGMCC 1.0892(Bacillussubtilis CGMCC 1.0892)和地衣芽孢桿菌 D3(Bacillus licheniformis D3)四種菌進行單菌和混菌發(fā)酵麩皮培養(yǎng)基,結(jié)果發(fā)現(xiàn)釀酒酵母菌與枯草芽孢桿菌混菌發(fā)酵效果最好,此時的麩皮多糖產(chǎn)量為44.55mg/g1.2接種比例及輔料的確定在發(fā)酵菌種及其組合確定的基礎(chǔ)上,以發(fā)酵麩皮中多糖產(chǎn)量作為指標,通過Plackett-Burman試驗設(shè)計和響應(yīng)面優(yōu)化法得到:釀酒酵母菌與枯草芽孢桿菌的最適接種比例為6.7:3.3;以麩皮為主料,豆粕粉和玉米粉為輔料的固態(tài)培養(yǎng)基配比為麩皮80.46%、豆粕粉9.32%和玉米粉10.22%。此時,發(fā)酵產(chǎn)物中多糖產(chǎn)量55.92mg/g1.3增效劑的確定在前面試驗的基礎(chǔ)上,以麩皮多糖產(chǎn)量作為指標,通過Plackett-Burman設(shè)計篩選MgS04、MnS04、CaCl2、KH2P04、尿素、檸檬酸鈉、酒石酸鉀鈉和腐植酸鈉等8種無機鹽和有機鹽,結(jié)果發(fā)現(xiàn)只有尿素對發(fā)酵麩皮多糖產(chǎn)量有顯著影響(P0.05),可作為發(fā)酵增效劑;對尿素添加量進行單因素優(yōu)化試驗得出:尿素的最佳添加量為0.1%。此時,發(fā)酵產(chǎn)物中多糖產(chǎn)量122.8 mg/g。1.4發(fā)酵條件的確定在前面試驗的基礎(chǔ)上,以多糖產(chǎn)量作為優(yōu)化指標,對發(fā)酵條件(發(fā)酵時間、發(fā)酵溫度、接種量、料水比)進行優(yōu)化。通過單因素和響應(yīng)面優(yōu)化試驗得到最后發(fā)酵條件為:發(fā)酵溫度35.4℃,發(fā)酵時間52.7h,接種量10.4%,料水比為1:1.16,此時得到發(fā)酵產(chǎn)物中實際多糖含量為130.21mg/g,較優(yōu)化前提高了 6%。1.5麩皮粗多糖提取條件研究由單因素試驗確定,浸提溫度80°C,浸提時間30min,料水比1:20為麩皮粗多糖的最佳提取條件。2麩皮多糖粗制品的抗氧化活性的研究通過腹腔注射Diquat建立氧化應(yīng)激動物模型,以研究麩皮多糖粗制品對Wistar大鼠氧化應(yīng)激的保護作用。試驗選用雄性Wistar大鼠36只,暫養(yǎng)后按試驗要求將大鼠隨機分為6組。低劑量組(low-dose,CL)、中劑量組(middle-dose,CM)和高劑量組(high-dose,CH):每天分別灌服100、200和400 mg/kg體重麩皮多糖粗制品;正對照組(positive control,PC):100mg/kg體重維生素C;負對照組(negative control,NC):空白攻毒;正常對照組(control,CT):空白不攻毒。試驗期14d。試驗結(jié)束當天,除CT組外,其它五組大鼠腹腔注射Diquat0.1mmol/kg體重,CT組注射等量生理鹽水,攻毒24h后屠宰。選取血漿、肝臟、腎臟、回腸和脾臟樣品,測定抗氧化指標和炎性因子指標。試驗表明:與NC組相比,麩皮多糖粗制品處理組的大鼠血漿和組織的CAT、SOD、GSH-Px活性和GSH含量顯著上升(P0.05),脂質(zhì)過氧化產(chǎn)物MDA水平顯著降低(P0.05),其中CH組大鼠的部分抗氧化指標恢復(fù)到CT組的水平;同時,麩皮多糖粗制品處理組的大鼠,血漿和肝臟、腎臟、回腸和脾臟的TNF-α、IL-6、IL-2和IL-1β炎性因子顯著低于NC組(P＜0.0S)。綜上,麩皮多糖粗制品可以有效緩解Diquat誘導(dǎo)的Wistar大鼠機體的氧化應(yīng)激。就整體效果而言,麩皮多糖粗制品的保護作用具有劑量依賴性。
[Abstract]:In this study, bran was used as the raw material, mixed with soybean meal powder, corn flour and other excipients, and used microbial fermentation to prepare the bran polysaccharide and study the antioxidant activity of its crude products. This paper has been studied in the following two aspects: 1 fermentation of bran polysaccharide by microorganism fermentation and extraction of 1.1 fermenting strains and the selection of their combinations to ferment bran P8 (LactobacillusplantarumP8), Saccharomyces cerevisiae CGMCC2.119 (Saccharomyces cerevisiae CGMCC2.119), Bacillus subtilis CGMCC 1.0892 (Bacillussubtilis CGMCC 1.0892) and Bacillus licheniformis D3 (Bacillus licheniformis) were used to ferment bran culture medium by bacteria and mixed bacteria. The results showed that the fermenting effect of Saccharomyces cerevisiae and Bacillus subtilis was the best. At this time, the yield of bran polysaccharide was 44.55mg/g1.2 inoculation ratio and the determination of the excipient materials on the basis of fermentation strain and its combination. The yield of polysaccharide in fermented bran was taken as an index, and the Plackett-Burman test design and response surface optimization were obtained. The optimum proportion of the inoculation of yeast and Bacillus subtilis is 6.7:3.3, and the ratio of solid medium with bran as the main material, soybean meal powder and corn flour is 80.46% of bran, 9.32% of soybean meal powder and corn flour 10.22%., and the determination of 55.92mg/g1.3 synergist in the production of polysaccharide in the fermentation product is based on the previous experiment with the production of Bran Polysaccharides. As an indicator, 8 kinds of inorganic salts and organic salts, such as MgS04, MnS04, CaCl2, KH2P04, urea, sodium citrate, potassium tartrate sodium and sodium humate, were designed and screened by Plackett-Burman. The results showed that only urea had a significant effect on the yield of fermented bran polysaccharide (P0.05), which could be used as a synergist for fermentation; the single factor optimization test of the urea addition was carried out. It is concluded that the optimum addition of urea is 0.1%., and the fermentation conditions of polysaccharide yield 122.8 mg/g.1.4 are determined on the basis of the previous experiment, and the optimization of the fermentation conditions (fermentation time, fermentation temperature, inoculation quantity and ratio of feed water) is carried out on the basis of the previous experiment, and the final hair is obtained by the single factor and the response surface optimization test. Fermentation conditions are: fermentation temperature 35.4 C, fermentation time 52.7h, inoculation amount 10.4%, material water ratio is 1:1.16, at this time the actual polysaccharide content in the fermentation product is 130.21mg/g, compared with the optimization before optimization, the extraction conditions of 6%.1.5 bran coarse polysaccharide were improved by single factor test, the extraction temperature was 80 C, the extraction time was 30min, and the material water was at 1:20 as the coarse polysaccharide of bran. The optimum extraction conditions of.2 bran polysaccharide crude products were studied by intraperitoneal injection of Diquat to establish an oxidative stress animal model to study the protective effect of bran on oxidative stress in Wistar rats. 36 male Wistar rats were selected, and the rats were randomly divided into 6 groups according to the test requirements. Low dose group (l Ow-dose, CL), medium dose group (middle-dose, CM) and high dose group (high-dose, CH): 100200 and 400 mg/kg weight Bran Polysaccharides each day respectively; positive control group (positive control, PC): 100mg/kg weight vitamin C; negative control group: blank attack; normal control group: blank no attack. Experimental period On the day of the experiment, in addition to group CT, other five groups of rats were injected with Diquat0.1mmol/kg weight intraperitoneally, group CT was injected with equal amount of saline, and 24h after 24h was slaughtered. Plasma, liver, kidney, ileum and spleen were selected to determine the antioxidant index and inflammatory factors. The experiment showed that compared with the NC group, the rat plasma of the bran polysaccharide crude processing group was plasma The activity of CAT, SOD, GSH-Px and GSH in the tissues increased significantly (P0.05), and the MDA level of lipid peroxidation decreased significantly (P0.05), and some of the antioxidant indices of group CH rats recovered to the level of group CT; meanwhile, the rats of the bran polysaccharide crude processing group, plasma and liver, kidney, ileum and spleen were TNF- a, IL-6, IL-2, and beta inflammatory properties. The factors were significantly lower than that in group NC (P < 0.0S). To sum up, bran polysaccharides can effectively alleviate the oxidative stress in Wistar rats induced by Diquat. As a whole, the protective effect of bran polysaccharide crude products is dose-dependent.
【學(xué)位授予單位】：內(nèi)蒙古農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S816

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