本文選題：絲瓜　切入點(diǎn)：多糖　出處：《沈陽(yáng)農(nóng)業(yè)大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：絲瓜,是一種常見的可供食用的植物,其化學(xué)成分復(fù)雜,含有多種活性成分。近年來(lái),植物多糖的研究成為眾人關(guān)注的焦點(diǎn),主要是由于植物多糖是一種天然的具有良好生物活性的物質(zhì),可用于治療多種疾病及提高人類的身心健康。然而對(duì)于絲瓜多糖的研究還處于初級(jí)階段,研究絲瓜多糖的性質(zhì)及結(jié)構(gòu)為日后絲瓜多糖產(chǎn)品的研發(fā)提供了理論依據(jù)。本文主要對(duì)絲瓜多糖的提取條件、分離純化方法、理化性質(zhì)、抗氧化活性及結(jié)構(gòu)進(jìn)行了研究,為擴(kuò)大絲瓜的使用范圍,研究絲瓜功能性食品提供參考依據(jù)。試驗(yàn)內(nèi)容與結(jié)果如下:(1)采用超聲波-酶復(fù)合法對(duì)絲瓜多糖進(jìn)行提取,利用響應(yīng)面法優(yōu)化絲瓜多糖提取工藝參數(shù)。以絲瓜多糖提取率為指標(biāo),對(duì)影響多糖提取率的四個(gè)因素:纖維素酶添加量、液料比、超聲溫度、超聲時(shí)間,進(jìn)行工藝參數(shù)優(yōu)化。在提取過(guò)程中,因素對(duì)絲瓜多糖提取率的影響大小順序?yàn)?液料比纖維素酶添加量超聲溫度超聲時(shí)間;提取工藝的優(yōu)化條件為:纖維素酶添加量5.6%,液料比33:1,超聲溫度72℃,超聲時(shí)間30min,該條件下得到的絲瓜多糖提取率為16.74%,說(shuō)明該方法能較好的用于絲瓜多糖提取條件的優(yōu)化。將得到的粗多糖命名為L(zhǎng)CP。(2)以絲瓜粗多糖LCP為原料,采用Sevage法脫蛋白,利用正交試驗(yàn)優(yōu)化脫蛋白條件。結(jié)果表明在V氯仿:V正丁醇比為3:1,VLCP:VSevage比為5:1,振蕩時(shí)間為20min的條件下脫蛋白效果最好,測(cè)得脫蛋白率為26.73%,多糖損失率為10.26%。(3)采用大孔樹脂法對(duì)脫蛋白后的絲瓜粗多糖LCP經(jīng)行了脫色試驗(yàn)。采用靜態(tài)吸附篩選出了大孔樹脂D301進(jìn)行動(dòng)態(tài)吸附試驗(yàn),最后結(jié)果表明上樣濃度3mg/mL,上樣量為10mL,上樣流速為2.5mL/min的條件下脫色效果最佳,得到的脫色率為74.05%,保留率為68.92%。(4)以脫蛋白脫色后的絲瓜粗多糖LCP為原料,采用DEAE-52纖維素柱層析,分別以蒸餾水、O.1mol/LNaCl溶液、0.2mol/LNaCl溶液、0.5mol/LNaCl溶液進(jìn)行洗脫得到LCPⅠ、LCPⅡ、LCPⅢ、LCPⅣ四個(gè)組分,收集多糖含量最高的組分LCPⅡ,多糖含量為63.53%。將LCPⅡ利用Sephadex G-75凝膠柱層析純化,以蒸餾水為洗脫液,得到單一的洗脫峰,收集濃縮、透析、凍干后得到LCPⅡ-1。經(jīng)紫外光譜掃描圖分析可知,LCPⅡ-1中幾乎不含蛋白質(zhì)、多肽、核酸等雜質(zhì)。(5)將絲瓜多糖LCPⅡ-1經(jīng)紅外光譜掃描可知,LCPⅡ-1具有多糖特征吸收峰,存在毗喃環(huán)結(jié)構(gòu)的單體。LCP Ⅱ-1的相對(duì)分子質(zhì)量為16766。(6)分離純化獲得絲瓜多糖LCPⅡ-1為白色絮狀固體,易溶于水,不溶于乙醇、乙醚、丙酮、乙酸乙酯等有機(jī)溶劑。經(jīng)化學(xué)反應(yīng)試驗(yàn)表明,該樣品中含有多糖組分,不含有蛋白質(zhì)、淀粉類物質(zhì)。體外抗氧化活性測(cè)定結(jié)果顯示,LCPⅡ-1對(duì)清除羥基自由基的能力較強(qiáng),絲瓜多糖濃度為1.0mg/mL時(shí),清除率最高為97.65%,和Vc相近;LCPⅡ-1對(duì)清除DPPH自由基的能力與Vc相比能力較弱,多糖濃度為10mg/mL時(shí),清除率達(dá)到最高為39.62%;清除超氧陰離子自由基能力不及Vc,但隨濃度增大呈上升趨勢(shì),當(dāng)濃度為5mg/mL時(shí),其清除率達(dá)到45.58%;LCPⅡ-1的總還原能力遠(yuǎn)不及Vc的還原能力,但在測(cè)量范圍內(nèi)呈上升趨勢(shì),具有一定的抗氧化能力。
[Abstract]:Luffa is a common edible plants, the complex chemical composition, contains many active ingredients. In recent years, the research of plant polysaccharides become the focus of attention, mainly because the plant polysaccharide is a natural substance with good biological activity, can be used for the treatment of various diseases and improve human the physical and mental health. However, for the study of Luffa polysaccharide is still at the primary stage, provides a theoretical basis for the research on the nature and structure of Luffa polysaccharide in the days after the gourd polysaccharide products. In this paper the extraction conditions of sponge gourd polysaccharide, methods of separation and purification, physicochemical properties, antioxidant activity and structure were studied, for to expand the scope of the use of sponge gourd, to provide reference for the research of Luffa functional food. The test contents and results are as follows: (1) the gourd polysaccharides were extracted by ultrasonic enzymatic method, optimization of Luffa using response surface method The extraction parameters in Luffa polysaccharide. The extraction rate of polysaccharide as the index, the four factors affecting the extraction rate of polysaccharides: the amount of cellulase, solid-liquid ratio, ultrasonic temperature, ultrasonic time, optimize the process parameters. In the extraction process, factors of Luffa polysaccharide extraction effect of the size of the order: liquid feed amount of ultrasonic temperature of ultrasonic time than cellulase; optimization of extraction conditions for cellulase addition 5.6%, solid-liquid ratio 33:1, ultrasonic temperature 72, ultrasonic time 30min, obtained the conditions of Luffa polysaccharide extraction rate is 16.74%, which shows that the method can optimize the extraction conditions of polysaccharide for Luffa better. The obtained crude polysaccharide named LCP. (2) in Luffa LCP polysaccharide as raw material, removal of protein by Sevage method, using orthogonal test conditions. The results show that the removal of protein in V chloroform: n-butanol V ratio of 3:1, VLCP: and VSevage ratio is 5:1, oscillation time Under the condition of 20min protein removal effect is the best, the measured protein removal rate was 26.73%, the polysaccharide loss rate was 10.26%. (3) by macroporous resin method for protein removal after the Luffa LCP crude polysaccharide were studied by static adsorption and decolorization test. Selected D301 macroporous resin by dynamic adsorption test, finally the results showed that the concentration of sample 3mg/mL, sample volume is 10mL, the optimum sample flow rate of decolorization effect under the condition of 2.5mL/min, the decolorization rate was 74.05%, the retention rate of 68.92%. (4) to remove protein after decolorization of Luffa LCP polysaccharide as raw material, using DEAE-52 cellulose chromatography respectively with distilled water. O.1mol/LNaCl solution, 0.2mol/LNaCl solution, 0.5mol/LNaCl solution was eluted with LCP I and LCP II, LCP III, LCP IV four components, collect the highest polysaccharide content components of LCP II, the polysaccharide content was 63.53%. LCP II by Sephadex G-75 gel column chromatography, distilled water As the eluent, elution peak, single collect concentration, dialysis and freeze-dried to obtain LCP II -1. by UV scan analysis, LCP II -1 almost no protein, polypeptide, nucleic acids and other impurities. (5) the gourd polysaccharide LCP II -1 by infrared spectroscopy scanning showed that LCP II -1 has the characteristic absorption peaks of polysaccharide monomer.LCP II and furan ring structure of the relative molecular weight of -1 was 16766. (6) purified from Luffa polysaccharide LCP II -1 white flocculent solids, soluble in water, insoluble in ethanol, ether, acetone, ethyl acetate and other organic solvents by. Reaction test showed that the samples containing polysaccharides, not contain protein, starch material. The determination results of antioxidant activity in vitro showed that LCP II -1 hydroxyl radical scavenging ability, gourd polysaccharide concentration is 1.0mg/mL, the clearance rate of up to 97.65%, and similar to Vc; scavenging DPPH free LCP II -1 Radical ability compared with Vc capacity is weak, the polysaccharide concentration is 10mg/mL, the clearance rate reached 39.62%; super anion free radical scavenging ability than Vc, but with the increase of concentration increased when the concentration of 5mg/mL, the removal rate reached 45.58%; LCP II -1 total reduction ability than the reduction ability of Vc, but the upward trend in the measurement range, has certain antioxidant capacity.

【學(xué)位授予單位】：沈陽(yáng)農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：TQ281

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