麸皮多糖微生物发酵制备及其粗制品抗氧化活性的研究

发布时间：2018-07-03 09:28

本文选题：麸皮多糖 + 微生物发酵　；参考：《内蒙古农业大学》2017年硕士论文

【摘要】：本研究以麸皮为原料,配以豆粕粉、玉米粉等辅料,利用微生物发酵法制备麸皮多糖并对其粗制品抗氧化活性进行研究。本论文从以下两个方面进行研究探讨:1麸皮多糖的微生物发酵制备和提取1.1发酵菌种及其组合的筛选以发酵麸皮中多糖产量为指标,利用植物乳杆菌P8(LactobacillusplantarumP8)、酿酒酵母 CGMCC2.119(Saccharomyces cerevisiae CGMCC2.119)、枯草芽孢杆菌CGMCC 1.0892(Bacillussubtilis CGMCC 1.0892)和地衣芽孢杆菌 D3(Bacillus licheniformis D3)四种菌进行单菌和混菌发酵麸皮培养基,结果发现酿酒酵母菌与枯草芽孢杆菌混菌发酵效果最好,此时的麸皮多糖产量为44.55mg/g1.2接种比例及辅料的确定在发酵菌种及其组合确定的基础上,以发酵麸皮中多糖产量作为指标,通过Plackett-Burman试验设计和响应面优化法得到:酿酒酵母菌与枯草芽孢杆菌的最适接种比例为6.7:3.3;以麸皮为主料,豆粕粉和玉米粉为辅料的固态培养基配比为麸皮80.46%、豆粕粉9.32%和玉米粉10.22%。此时,发酵产物中多糖产量55.92mg/g1.3增效剂的确定在前面试验的基础上,以麸皮多糖产量作为指标,通过Plackett-Burman设计筛选MgS04、MnS04、CaCl2、KH2P04、尿素、柠檬酸钠、酒石酸钾钠和腐植酸钠等8种无机盐和有机盐,结果发现只有尿素对发酵麸皮多糖产量有显著影响(P0.05),可作为发酵增效剂;对尿素添加量进行单因素优化试验得出:尿素的最佳添加量为0.1%。此时,发酵产物中多糖产量122.8 mg/g。1.4发酵条件的确定在前面试验的基础上,以多糖产量作为优化指标,对发酵条件(发酵时间、发酵温度、接种量、料水比)进行优化。通过单因素和响应面优化试验得到最后发酵条件为:发酵温度35.4℃,发酵时间52.7h,接种量10.4%,料水比为1:1.16,此时得到发酵产物中实际多糖含量为130.21mg/g,较优化前提高了 6%。1.5麸皮粗多糖提取条件研究由单因素试验确定,浸提温度80°C,浸提时间30min,料水比1:20为麸皮粗多糖的最佳提取条件。2麸皮多糖粗制品的抗氧化活性的研究通过腹腔注射Diquat建立氧化应激动物模型,以研究麸皮多糖粗制品对Wistar大鼠氧化应激的保护作用。试验选用雄性Wistar大鼠36只,暂养后按试验要求将大鼠随机分为6组。低剂量组(low-dose,CL)、中剂量组(middle-dose,CM)和高剂量组(high-dose,CH):每天分别灌服100、200和400 mg/kg体重麸皮多糖粗制品;正对照组(positive control,PC):100mg/kg体重维生素C;负对照组(negative control,NC):空白攻毒;正常对照组(control,CT):空白不攻毒。试验期14d。试验结束当天,除CT组外,其它五组大鼠腹腔注射Diquat0.1mmol/kg体重,CT组注射等量生理盐水,攻毒24h后屠宰。选取血浆、肝脏、肾脏、回肠和脾脏样品,测定抗氧化指标和炎性因子指标。试验表明:与NC组相比,麸皮多糖粗制品处理组的大鼠血浆和组织的CAT、SOD、GSH-Px活性和GSH含量显著上升(P0.05),脂质过氧化产物MDA水平显著降低(P0.05),其中CH组大鼠的部分抗氧化指标恢复到CT组的水平;同时,麸皮多糖粗制品处理组的大鼠,血浆和肝脏、肾脏、回肠和脾脏的TNF-α、IL-6、IL-2和IL-1β炎性因子显著低于NC组(P＜0.0S)。综上,麸皮多糖粗制品可以有效缓解Diquat诱导的Wistar大鼠机体的氧化应激。就整体效果而言,麸皮多糖粗制品的保护作用具有剂量依赖性。
[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.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S816

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