红豆越橘主要抑菌成分的研究

发布时间：2018-03-28 00:28

  本文选题：红豆越橘　切入点：总黄酮　出处：《哈尔滨商业大学》2017年硕士论文

【摘要】：红豆越橘中含丰富的有机酸、维生素、糖类和微量元素,还含有花青素、原花青素、黄烷醇、黄酮醇及其糖苷等活性成分,使红豆越橘具有抗氧化、抗癌、抵抗炎症等作用。目前,对红豆越橘生物活性的研究较少,其主要的开发和利用集中在果酒、饮料和果茶方面。本文主要以红豆越橘为研究对象,考究了它的抑菌成分,以及对不同微生物的抑制效果,同时进一步研究红豆越橘主要抑菌成分的抑菌机理,为红豆越橘进一步开发利用提供相关的理论依据。首先,以水和乙醇作为提取溶剂,对红豆越橘中的成分进行提取,经过初步的检测实验,证实了红豆越橘的提取液中含有氨基酸、蛋白质、糖类、酚类成分、黄酮及其甙类、生物碱、鞣质和蒽醌及其甙类等物质。并通过滤纸片法对红豆越橘提取物进行了初步的抑菌试验,后又采用对倍稀释法测得了红豆越橘水、乙醇提取物对供试菌的最低抑菌浓度(MIC)。红豆越橘水提取物对各供试菌的抑菌圈直径分别为:大肠杆菌17.3mm、枯草芽孢杆菌11.6mm、黑曲霉5.4mm和青霉6.0mm;MIC分别为:大肠杆菌31.25mg/mL、枯草芽孢杆菌62.5mg/mL、黑曲霉250mg/mL和青霉125mg/mL。红豆越橘乙醇提取物对各供试菌的抑菌圈直径分别为:枯草芽孢杆菌27mm,大肠杆菌20.6mm、黑曲霉5.9mm、青霉6.3mm;MIC分别为:大肠杆菌31.25mg/mL、枯草芽孢杆菌15.625mg/mL、黑曲霉125mg/mL和青霉62.5mg/mL。可见红豆越橘的乙醇提取物对供试菌的抑制作用强于水提取物。采用单因素和正交实验确定红豆越橘中总黄酮提取最佳条件为:超声波功率为450W、超声波处理时间为25min、乙醇水溶液浓度为50%、料液比为1:15,此条件下总黄酮提取率为29.87mg/g。当红豆越橘中总黄酮浓缩液的浓度为39.83mg/mL时对各供试菌的抑菌圈分别为:枯草芽孢杆菌21.67mm、大肠杆菌21.17mm、黑曲霉5.81mm和青霉6.10mm;MIC分别为:大肠杆菌5mg/mL、枯草芽孢杆5mg/mL、黑曲霉20mg/mL和青霉20mg/mL。而经中和后的红豆越橘总黄酮浓缩液(pH=7.0)对各菌株的MIC分别为:大肠杆菌400mg/mL、枯草芽孢杆菌600mg/mL,对黑曲霉和青霉没有明显抑制作用。采取响应曲面的设计方法,明确了红豆越橘中有机酸的最佳提取工艺:超声波功率150W,超声波处理温度70℃,溶剂浓度为10%的乙醇水溶液,料液比为1:25,提取时间为40min。当其有机酸提取浓缩液的浓度为1.275g/mL时,有机酸对大肠杆菌的MIC为7.969mg/mL、枯草芽孢杆的MIC为15.938mg/mL、黑曲霉的MIC为63.75mg/mL和青霉的MIC为31.875mg/mL。通过超微结构观察和生化等试验,研究红豆越橘有机酸浓缩液的抑菌机理。有机酸浓缩液导致供试菌的离子及小分子物质漏出;各供试菌菌体对还原糖的利用能力下降、可溶性蛋白质大量溶出;细胞表面出现褶皱变形,部分出现溶胀破裂。
[Abstract]:Red bean blueberries are rich in organic acids, vitamins, carbohydrates and trace elements, as well as anthocyanins, proanthocyanidins, flavanols, flavonols and their glycosides. At present, there are few studies on the biological activity of red bean blueberry, the main development and utilization of which are concentrated on wine, beverage and tea. In this paper, red bean blueberry is taken as the object of study, and its antimicrobial composition is studied. At the same time, the inhibition mechanism of main antimicrobial components of red bean blueberry was further studied, which provided relevant theoretical basis for further development and utilization of red bean blueberry. Firstly, water and ethanol were used as extraction solvent. The components of red bean blueberry were extracted. The results showed that the extract of red bean blueberry contained amino acids, proteins, carbohydrates, phenols, flavonoids and its glycosides, alkaloids. Tannin, anthraquinone and its glycosides, etc. Preliminary bacteriostatic test was carried out on the extract of red bean blueberry by filter paper method, and then the red bean blueberry water was determined by double dilution method. The minimum inhibitory concentration (MIC) of ethanol extract on the tested bacteria was: E.coli 17.3mm, Bacillus subtilis 11.6mm, Aspergillus Niger 5.4mm and penicillium 6.0mmmL, respectively. Bacillus subtilis 62.5 mg / mL, Aspergillus Niger 250mg/mL and Penicillium 125mg / mL. The inhibitory circle diameters of ethanol extract of red bean blueberry against each tested bacterium were: Bacillus subtilis 27mm, Escherichia coli 20.6mm, Aspergillus Niger 5.9mm, Penicillium 6.3mmMIC: E.coli 31.25mg / mL, Bacillus subtilis L. Bacilli 15.625 mg / mL, Aspergillus Niger 125mg/mL and Penicillium 62.5 mg / mL. It can be seen that ethanol extract of red bean blueberry has stronger inhibitory effect on tested bacteria than water extract. Single factor and orthogonal experiment were used to determine the best conditions for extracting total flavonoids from red bean blueberry: ultrasonic wave. The power is 450 W, the ultrasonic treatment time is 25 min, the concentration of ethanol solution is 50 min, the ratio of material to liquid is 1: 15, and the extraction rate of total flavonoids is 29.87 mg / g. When the concentration of total flavone concentrate in blueberry is 39.83mg/mL, the inhibition zone of each tested bacteria is as follows:. The mics of Bacillus subtilis 21.67 mm, Escherichia coli 21.17 mm, Aspergillus Niger 5.81mm and Penicillium 6.10 mm-1 were Escherichia coli 5 mg / mL, Bacillus subtilis 5 mg / mL, Aspergillus Niger 20mg/mL and Penicillium penicillium 20 mg / mL. The MIC of each strain was:. Escherichia coli 400 mg / mL, Bacillus subtilis 600 mg / mL, has no significant inhibition on Aspergillus Niger and Penicillium. The optimum extraction technology of organic acid from red bean blueberry was determined as follows: ultrasonic power 150W, ultrasonic treatment temperature 70 鈩，

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