本文選題：紅豆越橘　切入點(diǎn)：總黃酮　出處：《哈爾濱商業(yè)大學(xué)》2017年碩士論文

【摘要】：紅豆越橘中含豐富的有機(jī)酸、維生素、糖類和微量元素,還含有花青素、原花青素、黃烷醇、黃酮醇及其糖苷等活性成分,使紅豆越橘具有抗氧化、抗癌、抵抗炎癥等作用。目前,對(duì)紅豆越橘生物活性的研究較少,其主要的開發(fā)和利用集中在果酒、飲料和果茶方面。本文主要以紅豆越橘為研究對(duì)象,考究了它的抑菌成分,以及對(duì)不同微生物的抑制效果,同時(shí)進(jìn)一步研究紅豆越橘主要抑菌成分的抑菌機(jī)理,為紅豆越橘進(jìn)一步開發(fā)利用提供相關(guān)的理論依據(jù)。首先,以水和乙醇作為提取溶劑,對(duì)紅豆越橘中的成分進(jìn)行提取,經(jīng)過初步的檢測(cè)實(shí)驗(yàn),證實(shí)了紅豆越橘的提取液中含有氨基酸、蛋白質(zhì)、糖類、酚類成分、黃酮及其甙類、生物堿、鞣質(zhì)和蒽醌及其甙類等物質(zhì)。并通過濾紙片法對(duì)紅豆越橘提取物進(jìn)行了初步的抑菌試驗(yàn),后又采用對(duì)倍稀釋法測(cè)得了紅豆越橘水、乙醇提取物對(duì)供試菌的最低抑菌濃度(MIC)。紅豆越橘水提取物對(duì)各供試菌的抑菌圈直徑分別為:大腸桿菌17.3mm、枯草芽孢桿菌11.6mm、黑曲霉5.4mm和青霉6.0mm;MIC分別為:大腸桿菌31.25mg/mL、枯草芽孢桿菌62.5mg/mL、黑曲霉250mg/mL和青霉125mg/mL。紅豆越橘乙醇提取物對(duì)各供試菌的抑菌圈直徑分別為:枯草芽孢桿菌27mm,大腸桿菌20.6mm、黑曲霉5.9mm、青霉6.3mm;MIC分別為:大腸桿菌31.25mg/mL、枯草芽孢桿菌15.625mg/mL、黑曲霉125mg/mL和青霉62.5mg/mL。可見紅豆越橘的乙醇提取物對(duì)供試菌的抑制作用強(qiáng)于水提取物。采用單因素和正交實(shí)驗(yàn)確定紅豆越橘中總黃酮提取最佳條件為:超聲波功率為450W、超聲波處理時(shí)間為25min、乙醇水溶液濃度為50%、料液比為1:15,此條件下總黃酮提取率為29.87mg/g。當(dāng)紅豆越橘中總黃酮濃縮液的濃度為39.83mg/mL時(shí)對(duì)各供試菌的抑菌圈分別為:枯草芽孢桿菌21.67mm、大腸桿菌21.17mm、黑曲霉5.81mm和青霉6.10mm;MIC分別為:大腸桿菌5mg/mL、枯草芽孢桿5mg/mL、黑曲霉20mg/mL和青霉20mg/mL。而經(jīng)中和后的紅豆越橘總黃酮濃縮液(pH=7.0)對(duì)各菌株的MIC分別為:大腸桿菌400mg/mL、枯草芽孢桿菌600mg/mL,對(duì)黑曲霉和青霉沒有明顯抑制作用。采取響應(yīng)曲面的設(shè)計(jì)方法,明確了紅豆越橘中有機(jī)酸的最佳提取工藝:超聲波功率150W,超聲波處理溫度70℃,溶劑濃度為10%的乙醇水溶液,料液比為1:25,提取時(shí)間為40min。當(dāng)其有機(jī)酸提取濃縮液的濃度為1.275g/mL時(shí),有機(jī)酸對(duì)大腸桿菌的MIC為7.969mg/mL、枯草芽孢桿的MIC為15.938mg/mL、黑曲霉的MIC為63.75mg/mL和青霉的MIC為31.875mg/mL。通過超微結(jié)構(gòu)觀察和生化等試驗(yàn),研究紅豆越橘有機(jī)酸濃縮液的抑菌機(jī)理。有機(jī)酸濃縮液導(dǎo)致供試菌的離子及小分子物質(zhì)漏出;各供試菌菌體對(duì)還原糖的利用能力下降、可溶性蛋白質(zhì)大量溶出;細(xì)胞表面出現(xiàn)褶皺變形,部分出現(xiàn)溶脹破裂。
[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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