【作者】 章文琴；

【导师】 刘成梅；

【作者基本信息】 南昌大学 ， 农产品加工与贮藏工程， 2010， 硕士

【摘要】 本课题采用热提-极性pH法从台湾75-1大豆中提取大豆多糖,并以动态高压微射流技术(dynamic high pressure microfluidization,DHPM)为手段对大豆可溶性多糖(soluble soybean polysaccharides,SSPS)进行改性,研究DHPM对SSPS组分、结构及功能特性的影响。1、大豆多糖分离纯化实验结果表明：选择台湾75-1大豆(糖含量28.2%)为实验对象,采用热提-极性pH值法提取SSPS,确定实验条件为：碱脱蛋白料液比1：10,pH值9.5；酸提SSPS提取温度120℃,pH4.0,得率为41.83%。使用离子交换树脂色谱分离法纯化多糖,确定上样量为50mg,缓冲液pH值为6.8。分级得到两个组分SSPS-1.SSPS-2,通过HPLC凝胶色谱和紫外谱图确定SSPS-2的分子量为91.5kD,蛋白含量86.7%,为高结合蛋白含量的单一多糖。2、SSPS-1组分的结构分析表明：SSPS-1含有5个单峰,相对分子量分别为733kD、295 kD、25 kD、6223D及1589D,含少量蛋白的杂多糖；SSPS-1经过GC、红外扫描及甲基化分析,确定其为β-吡喃糖,且含有较多糖醛酸,主链为鼠李糖与半乳糖醛酸,连接方式为1-3-Rha及1-4-GalA,其他组成糖为阿拉伯糖、半乳糖、木糖及甘露糖等。3、DHPM对SSPS-1结构的影响：对SSPS-1进行DHPM处理,大分子量组分降低,小分子量组分降低,中等分子量组分增加,分析DHPM对SSPS-1的作用方式可能为：大分子向低分子量转化,低分子量向高分子量聚集。SSPS-1由原来的针状堆积排列,向末端膨大呈球形的“火柴棒”状有序排列结构转变。经DHPM处理的SSPS-1主链的单糖Rha和GalA含量降低,侧链部分的单糖Ara、Gal、Fuc、Man降低,Xyl、Glc、GlcA则未检出。连接主链的糖苷键(1→3)键鼠李糖及(1→4)键半乳糖醛酸含量降低,而其主要支链上的糖苷键的含量得到增加。DHPM处理会增加糖醛酸含量,但糖环构型未改变。4、DHPM对SSPS-1理化性质的影响：SSPS-1随着DHPM的处理次数的增加,粒径分布、Zeta电位、表观粘度,均呈现先降低,而后回升,最后降低的趋势。SSPS-1经DHPM 150MPa处理4次后的乳化性、乳化稳定性呈现出最大值。SSPS-1经处理6次后,起泡性达到最大值,SSPS-1的起泡稳定性在处理2次后达到最高峰。5、DHPM对SSPS-1生理功能的影响：健康小鼠喂食经DHPM处理后的SSPS-1,其粪便菌群及肠道菌群中的双歧杆菌数目和乳酸菌的数目均有所增加,大肠杆菌数目下降,即SSPS-1对有益菌(双歧杆菌、乳酸菌)有增殖效果,对有害菌(大肠杆菌)有抑制效果。SSPS-1对两种有益菌的增加比例相当,且高于有害菌的抑制比例。SSPS-1经DHPM 150MPa处理4次后,对有益菌的增殖比例最高,对大肠杆菌的抑制比例亦最高。SSPS-1对胆汁酸有一个吸附解析的过程,DHPM的作用使得SSPS-1对胆汁酸的吸附量增加,胆汁酸的释放延长。SSPS-1对Ca2+、Mg2+是一个动态平衡过程,SSPS-1对Cu2+、Pb2+的吸附相对稳定,呈缓慢释放的现象。综上所述：DHPM的作用可使分级得到的SSPS分子排布向中等分子量靠拢,进一步改变糖苷键比例、单糖组成、官能团,从而使SSPS的粒径分布、表观粘度及乳化起泡能力,对健康小鼠肠道、粪便菌群影响,对胆汁酸及Ca2+、Mg2+、Cu2+、Pb2+等金属离子的吸附能力改变。更多还原

【Abstract】 Dynamic high pressure microfluidization (DHPM) treated on soybean soluble polysaccharides (SSPS) composition, structure and functional properties, is good for understanding the molecular structure in the role of the polysaccharide to DHPM mechanism, and comprehending SSPS diversifications in relationships between structure and function.1、The soybean purification experimental results showed:Taiwan 75-1 soybean (28.2% sugar content) was selected to be our experimental subjects. SSPS was extracted by high temperature-polar pH value method (compared to enzymatic method).The main points of extraction:alkali deproteinized procedure:liquid ratio 1:10, pH value 9.5; acid extracting SSPS procedure:temperature 120℃, pH4.0.The total yield is 41.83 percent. Ion exchange resin chromatography was used to purify polysaccharide, determining the sample volume was 50mg, buffer pH value is 6.8. Classification of the SSPS obtained two components of SSPS-1, SSPS-2 by detecting the molecular weight spectra and UV spectra, SSPS-2 molecular weight is 91.5kD, protein content is 86.7%,which was the high-binding protein content in single polysaccharide. SSPS-1 was conducted as the object of DHPM treatment.2、SSPS-1 contains five single peak, and the relative molecular weights are 733kD,295 kD, 25 kD,6223D and 1589D respectively, which was considered to be the complex polysaccharides containing a small amount of protein.lt was determined to be theβ-pyranose, and the main chain involved of rhamnose and galacturonic acid, which was connected by 1-3-Rha, and 1-4-GalA.Other components of the polysaccharides were arabinose, galactose, wood glucose and mannose through the GC, infrared scanning and methylation analysis.3、The analysis of the components from SSPS-1:dealed with DHPM processing, large molecular weight components of SSPS-1 decreased,and small molecular weight components lowed, with middle molecular weight fractions increased.The mode of DHPM process on SSPS-1: macromolecules intended to lower molecular weight while low molecular weight converted to high molecular weight aggregates. SSPS-1 changed into a spherical enlarged end of the "match stick"-like structure of ordered arrangement from the needle-like stacking arrangement.The main chain monosaccharides content of SSPS-1 treated by DHPM:Rha and GalA,dropped down and part of the single-sugar side chain such as Ara, Gal, Fuc and Man reduced, and accompanily Xyl, Glc, GlcA was not detected. The content of Glycosidic bonds connecting the main chain of (1→3) Rha and (1→4) GalA decreased, while the content of glycosidic bonds connecting the main branch chain increased. Generally DHPM processing will increase the uronic acid content, but the polysaccharides ring configuration has not changed.4、The physical and chemical Properties of SSPS-1 treated by DHPM:SSPS-1 treated by DHPM with the increase of dealing times,its size distribution, Zeta potential, apparent viscosity, were decreased first, then showed a small peak. The emulsification, emulsion stability of SSPS-1 treated by DHPM processing at 150MPa、4 times showed the maximum value. SSPS-1 treated by six times, reached the highest position of foaming capacity while foaming stability treated by 2 times increased to the peak.5、The physiological function of SSPS-1 treated by DHPM:with DHPM treatment on the SSPS-1, the bifidobacterium flora and lactic acid bacteria have increased, followed by the number of E. coli decreased in the fecal flora and intestinal bacteria of the healthy mouse. In another way,SSPS-1 increased the beneficial bacteria (bifidobacterium bacteria, lactic acid bacteria) in a multiplier effect, and decreased the harmful bacteria (E. coli) an inhibitory effects. The two beneficial bacteria increased ratio effected by SSPS-1 is higher than the percentage inhibition of harmful bacteria. SSPS-1 treated by DHPM processing at 150MPa、4 times, have an effect on beneficial bacteria with the highest proportion and the inhibition of E. coli with the highest proportion.The absorption of bile acid on SSPS-1, was a process of adsorption analysis. The treatment of DHPM to SSPS-1 on bile acid adsorption added to the amount of absorption, simultaneously extended the release time of bile acids. The adsorption of SSPS-1 on Ca2+, Mg2+ is a dynamic equilibrium process, whlie the process on Cu2+, Pb2+ adsorption is a relatively stable, slow-released phenomenon.In summary:the treatment of DHPM can make the higher and lower molecular weight of classified SSPS(SSPS-1) moved closer to the medium molecular weight.Further the SSPS-1 changed in the glycosidic bond ratio, the monosaccharide composition and the functional groups, resulting the diversification of its functional properties and physiological properties (the size distribution, the apparent viscosity,the foaming capacity and foaming stability, emulsifying capacity and emulsifying stability,the fecal flora and intestinal bacteria from a healthy mice, the bile acids and Ca2+, Mg2+, Cu2+, Pb2+ metal ions adsorption capacity).更多还原