烟管菌多糖的分离制备及降血糖活性研究

发布时间：2018-05-29 23:25

本文选题：多糖 + 响应面　；参考：《沈阳农业大学》2017年硕士论文

【摘要】：烟管菌属多孔菌科真菌,有关烟管菌生物学效应的报道多涉及其抗癌活性物质,而烟管菌活性多糖降血糖作用则较少报道。本研究以本实验室分离的多孔菌科烟管菌为实验材料,对其种属鉴定、活性多糖提取及降血糖作用进行研究,主要结果如下:1.通过对真菌进行ITS测序,经BLAST比对鉴定菌种为担子菌纲非褶菌目多孔菌科烟管菌属烟管菌。2.以菌丝干重和多糖含量为标准,其中以多糖含量为主,筛选出的最适合的液体发酵培养基组分为:乳糖20g,酵母浸粉3.5g,MgS04.7H2O1g,KH2P042g,去皮马铃薯 100 g,H20 1 L。3.以菌丝干重为标准,筛选出的最适的发酵条件为:pH 6,培养时间8 d,培养温度 27℃转速160r·min-1 接种量 8mL,装液量 100mL/250mL。4.为了探索超声波辅助提取烟管菌菌丝体多糖的最佳工艺,在单因素试验的基础上选取水料比、超声时间、超声温度三个因素为影响因子,利用响应面Box-Behnken中心组合试验的范围和水平,以烟管菌菌丝体多糖提取率为响应值,进一步优化了超声波辅助提取多糖的条件。通过研究和探讨三个因素的最佳水平范围和相互之间的作用,建立的影响菌丝体多糖提取率的模型为Y = 8.56-0.20X1-0.25X2 + 0.27X3 + 0.080X1X2 + 0.010X1X3-0.025X2X3-0.66X12-0.54X22-0.42X32。由统计学分析结果可以知道,方程中一次项X1、X2、X3,二次项X12、X22、X23对多糖提取率有非常显著的影响(p0.01),并且交互项X1X2对多糖提取率有显著(p0.05)的影响。根据回归分析结果得到的超声波辅助提取烟管菌菌丝体多糖的最佳工艺条件为:水料比48:1,超声时间27 min,超声温度53℃。在此条件下菌丝体多糖提取率达到8.57%。5.将烟管菌多糖配制成不同浓度水溶液,测定各自的DPPH清除率,在浓度为1.5 mg·mL-1时,DPPH清除率可以达到73.39%。6.用IBMX+DEX+Insulin联合的方法将3T3-L1前脂肪细胞成功诱导为脂肪细胞,选择3μmol/LDEX对进行脂肪细胞进行诱导,建立了胰岛素抵抗(IR)细胞模型,经过研究发现烟管菌菌丝体多糖具有降糖活性。7.为了对多糖分子形貌进行表征,分别对多糖进行拉曼光谱和扫描电镜。采用532 nm激光光源,测试了烟管菌多糖的拉曼光谱。光谱中出现了 440 cm-1峰表明多糖中含有葡萄糖。而通过对多糖进行扫描电镜,在不同倍数物镜下的烟孔菌多糖呈现不同形态。200 μm物镜下可以看到多糖为枯叶状,表面粗糙;在50 μm物镜下则呈不规则的碎石状,而在5 μm物镜下多糖主要呈规则的蚕豆、小球形态,表面平滑。
[Abstract]:There are many reports about the biological effects of tobacco tube bacteria on the biological effects of the bacteria, and the antitumor activity of the bacteria is less reported. The results are as follows: 1. by ITS sequencing of fungi and BLAST comparison, the strain was identified as the strain.2. of the genus basidiomycetes of the genus polyporia. The dry weight of mycelium and the content of polysaccharide were the standard, and the most suitable liquid fermentation culture medium was divided into lactose 20g, yeast extract 3.5g, MgS04.7H2O1. G, KH2P042g, peeled potato 100 g, H20 1 L.3. were the standard of mycelial dry weight. The optimum fermentation conditions were: pH 6, incubation time 8 D, 160r. Min-1 inoculation at 27 centigrade temperature, 8mL, and liquid volume 100mL/250mL.4. in order to explore the optimum process of ultrasonic assisted extraction of mycelium polysaccharides from the mycelium of tobacco tube bacteria, in the basis of single factor test On the basis of the selection of water material ratio, ultrasonic time and ultrasonic temperature three factors as the influencing factors, using the range and level of the response surface Box-Behnken center combination test, the conditions of ultrasonic assisted extraction of polysaccharide were further optimized with the response value of the polysaccharide extraction rate of mycelium of the mycelium of the tobacco tube bacteria. The optimum level range of three factors was studied and discussed. The interaction between Y = 8.56-0.20X1-0.25X2 + 0.27X3 + 0.080X1X2 + 0.010X1X3-0.025X2X3-0.66X12-0.54X22-0.42X32. was established by the model of influencing the polysaccharide extraction rate of mycelium. The one term X1, X2, X3, two term X12, X22, and X23 had a very significant effect on the extraction rate of polysaccharide (P0.01). And the interaction term X1X2 has a significant effect on the extraction rate of polysaccharide (P0.05). The optimum technological conditions for ultrasonic assisted extraction of Polysaccharide from the mycelium of the mycelia obtained by the regression analysis are as follows: the ratio of water to 48:1, the ultrasonic time 27 min, and the ultrasonic temperature of 53. Under this condition, the polysaccharide extraction rate of mycelium is 8.57%.5. and the polysaccharide of the tobacco tube bacteria is prepared. The DPPH scavenging rate of different concentrations of water was measured. When the concentration was 1.5 mg. ML-1, the DPPH clearance rate could be achieved by 73.39%.6. using IBMX+DEX+Insulin to induce the 3T3-L1 preadipocytes to be successfully induced as adipocytes, and 3 u mol/LDEX were selected to induce the adipocytes, and the insulin resistance (IR) cell model was established. It was found that the polysaccharides of the mycelium of the mycelium were characterized by the hypoglycemic activity of.7. in order to characterize the molecular morphology of polysaccharides. The Raman and scanning electron microscopy of polysaccharide were carried out respectively. The Raman spectra of the polysaccharide were measured with 532 nm laser light source. The peak of 440 cm-1 in the spectrum showed that the glucose was contained in the polysaccharide. Under the microscope, the polysaccharides under different multiple objective lenses showed different forms of.200 mu m objective. The polysaccharides could be seen as dry leaves and rough surface. Under the 50 mu m objective lens, the irregular macadam was found. Under the 5 m objective lens, the polysaccharide mainly showed the regular broad bean, the shape of the ball and the surface was smooth.
【学位授予单位】：沈阳农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R284;R285

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