本文選題：魔芋飛粉　切入點(diǎn)：ACE抑制肽　出處：《陜西科技大學(xué)》2017年碩士論文

【摘要】：血管緊張素轉(zhuǎn)化酶(Angiotensin I-Converting Enzyme,ACE)抑制肽是一種對(duì)ACE活性具有抑制作用的多肽,一般由2~20個(gè)氨基酸殘基組成。ACE抑制肽通過與ACE發(fā)生競(jìng)爭性結(jié)合,能阻斷血管緊張素Ⅱ生成、促進(jìn)舒緩肽和腦啡肽生成,從而起到降壓作用。目前,已有從綠豆、大豆、酪蛋白、牛乳、麥胚蛋白、牡蠣等原料中制備ACE抑制肽的報(bào)道,而以魔芋為原料制備ACE抑制肽的資料甚少。檢測(cè)結(jié)果發(fā)現(xiàn),富含蛋白的魔芋飛粉經(jīng)過酶法降解可產(chǎn)生支鏈氨基酸寡肽、ACE抑制肽、高F值寡肽、甘露聚糖肽等活性肽。對(duì)魔芋飛粉這一天然藥物資源進(jìn)行研究開發(fā),有望為ACE抑制肽的制備開辟一條綠色的新途徑。本文以魔芋飛粉為原料,采用酶解法制備魔芋ACE抑制肽,依次通過超濾及大孔吸附樹脂對(duì)制備的ACE抑制肽分離純化,并對(duì)其進(jìn)行質(zhì)量檢測(cè)及理化性質(zhì)分析。研究結(jié)果如下:1.魔芋ACE抑制肽酶解法制備:以ACE抑制率為指標(biāo),通過對(duì)預(yù)實(shí)驗(yàn)篩選出的胃蛋白酶酶解制備魔芋ACE抑制肽的工藝進(jìn)行單因素考察和正交優(yōu)化,得到最佳酶解工藝條件為:底物濃度2%、酶用量5000U/g、溫度37℃、pH2.5、酶解時(shí)間210min,此時(shí)酶解液的ACE抑制率為92.65%、多肽得率為12.60%。2.超濾法分離魔芋ACE抑制肽:以ACE抑制率為指標(biāo),預(yù)選出最佳超濾膜的截留分子量為1kDa。單因素考察及正交實(shí)驗(yàn)優(yōu)化得到的最佳超濾條件為:在多肽液濃度2%、p H4.0、溫度40℃的條件下,采用1kDa的超濾膜超濾15min,產(chǎn)物的ACE抑制率可達(dá)到92.75%。3.大孔吸附樹脂分離魔芋ACE抑制肽:以吸附率和解吸率為指標(biāo),通過對(duì)預(yù)選出的DA201-C型大孔吸附樹脂進(jìn)行吸附-解吸工藝實(shí)驗(yàn),確定出最佳吸附工藝條件為:上樣液質(zhì)量濃度8mg/mL、p H2.0、上樣量8mL、上樣流速0.5mL/min;解吸工藝條件為:洗脫液乙醇體積分?jǐn)?shù)80%、洗脫流速1.0mL/min、洗脫時(shí)間2h。在此條件下,得到的魔芋ACE抑制肽的抑制率為95.19%。4.紅外光譜分析:對(duì)分離純化后的魔芋ACE抑制肽及ACE抑制肽標(biāo)準(zhǔn)品進(jìn)行紅外光譜掃描,結(jié)果表明,二者的峰形和峰值大體一致,均存在O-H、N-H、酰胺鍵、C-O、芳烴的C-H彎曲振動(dòng)等。綜合分析結(jié)果,魔芋ACE抑制肽與標(biāo)準(zhǔn)品紅外光譜特征基本一致。5.采用紫外全波長掃描對(duì)魔芋ACE抑制肽進(jìn)行檢測(cè),結(jié)果在220nm處有最大吸收峰,與ACE抑制肽標(biāo)準(zhǔn)品基本一致。高效液相色譜法分析結(jié)果表明,魔芋ACE抑制肽與標(biāo)準(zhǔn)品峰形特征相似。紫外分光光度法檢測(cè)出純化后樣品中魔芋ACE抑制肽含量為81.37%。凝膠滲透色譜法測(cè)定出魔芋ACE抑制肽的峰值分子量(MP)為655Da,分子量范圍為500~700Da。6.魔芋ACE抑制肽理化性質(zhì)分析:分離純化后得到的魔芋ACE抑制肽為灰白色固體粉末,有腥臭味;室溫下pH2.0時(shí)在水中的溶解率可達(dá)95%左右;30~50℃下穩(wěn)定性較好,但90℃水浴下5h后其ACE抑制率幾乎為零;在pH2.0~4.0范圍內(nèi)室溫放置2h ACE抑制活性無明顯變化,但隨著p H值逐漸增大,ACE抑制活性會(huì)逐漸降低;NaCl濃度為0.2~0.6mol/L時(shí)室溫放置2h ACE抑制率仍能維持在90%左右;37℃下魔芋ACE抑制肽在消化酶中穩(wěn)定性較好,ACE抑制率基本維持在85%。
[Abstract]:Angiotensin converting enzyme (Angiotensin I-Converting Enzyme, ACE) inhibitory peptide is a polypeptide with inhibitory effect on the activity of ACE, consists of 2~20 amino acid residues of.ACE inhibitory peptides through the combination of competition and ACE, can block angiotensin, promote soothing peptides and enkephalin generated thereby to the hypotensive effect. At present, the existing from mung bean, soybean, casein, milk, wheat germ protein, preparation of ACE inhibitory peptides from oyster and other raw materials reported, with Konjac as raw materials for preparation of ACE inhibitory peptides with little data. The results show that the rich protein containing konjac powder after enzymatic degradation can produce branched amino acid peptide, ACE inhibitory peptide, high F-measure oligopeptide, mannan peptide active peptides. The konjac powder the natural medicine resources for research and development, is expected for the preparation of ACE inhibitory peptides to open up a new way to green. In this paper, konjac powder As raw materials, preparation of ACE inhibitory peptides by enzyme hydrolysis method of konjac, followed by ultrafiltration and macroporous resin on the preparation of ACE inhibitory peptides were isolated and purified, and carries on the analysis and physicochemical properties of quality detection. The results are as follows: 1. konjac ACE inhibitory peptides prepared by enzymatic hydrolysis with ACE inhibition rate as the index. The pepsin of pre experiment screened the solution preparation process of konjac ACE inhibitory peptides of single factor test and orthogonal optimization, the optimal enzymatic hydrolysis conditions: substrate concentration of 2%, enzyme dosage 5000U/g, temperature 37 C, pH2.5, hydrolysis time 210min, the enzymolysis liquid of ACE inhibition rate was 92.65%. The yield of polypeptide 12.60%.2. ultrafiltration separation of konjac ACE inhibitory peptides with ACE inhibitory activity as index, the optimum selection of ultrafiltration membrane MWCO 1kDa. single factor and orthogonal experiment to optimize the best ultrafiltration conditions: in peptide concentration of 2%, P H4.0, temperature 40 degrees C under the condition of using 1kDa ultrafiltration membrane ultrafiltration 15min, the inhibition rate of the product ACE can achieve the separation of konjac ACE inhibitory peptides by 92.75%.3. macroporous resin adsorption rate and desorption rate as the index, the adsorption of DA201-C macroporous resin pre selected desorption experiment, the optimum adsorption conditions: the concentration of sample solution 8mg/mL, P H2.0, sample volume 8mL, sample flow rate 0.5mL/min; desorption conditions: eluent ethanol concentration 80%, elution rate 1.0mL/min, elution time 2h. under these conditions, the inhibition rate of ACE peptide was konjac 95.19%.4. infrared spectroscopy analysis of separation purified konjac ACE inhibitory peptides and ACE inhibitory peptides standard IR scanning results showed that two of the peak and shape of roughly the same, there are O-H, N-H, C-O, aromatic amide bond, the C-H bending vibration. The comprehensive analysis. Fruit, konjac ACE inhibition characteristics of infrared spectra of peptides with the standard is consistent with.5. UV wavelength scanning of konjac ACE inhibitory peptides were detected in the 220nm has the maximum absorption peak, consistent with the ACE inhibitory peptide standard. HPLC analysis showed that konjac ACE inhibitory peptides with standard peak similar characteristics. The ultraviolet spectrophotometry to detect the purified samples of konjac ACE inhibitory peptide content of 81.37%. gel permeation chromatography method for the determination of the peak molecular weight of ACE inhibitory peptide of konjac (MP) for 655Da, the molecular weight range of 500~700Da.6. konjac ACE inhibitory peptide physicochemical properties analysis: purified konjac ACE inhibition the peptide is grey white powder, have xingchouwei; rate of up to 95% solubility in water at room temperature at about 30~50 DEG pH2.0; good stability, but under 90 DEG C water bath after 5h the ACE inhibition rate is almost zero; in the range of 4 pH2.0~ Room temperature placed 2H ACE inhibitory activity had no obvious change, but with the P H increases, ACE inhibitory activity will gradually decrease; when the concentration of 0.2~0.6mol/L NaCl 2H ACE inhibition rate at room temperature can be maintained at about 90% DEG C; 37 konjac ACE inhibitory peptide had better stability of digestive enzymes, the inhibition rate of ACE remained at 85%.

【學(xué)位授予單位】：陜西科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類號(hào)】：R945

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相關(guān)期刊論文 前10條

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本文編號(hào)：1708304