酶解鸡蛋清小（寡）肽混合物对小鼠免疫功能的影响及其机理研究
发布时间：2018-08-20 10:32
【摘要】：为了初步探索酶解鸡蛋清蛋白产生的小(寡)肽混合物是否具有免疫调节作用及其调节机体免疫功能的机理，本研究在筛选适宜酶解条件并制备酶解鸡蛋清小(寡)肽混合物基础上，考察了不同鸡蛋清小(寡)肽混合物对健康小鼠体外免疫功能的影响，选出免疫增强效果较优的小(寡)肽混合物，研究其对免疫功能正常和免疫力低下小鼠体内免疫功能的影响，并初步探索鸡蛋清小(寡)肽混合物影响机体免疫功能的机理。本研究包括以下四个部分。 第一部分 蛋白酶酶解条件筛选 试验一 7种蛋白酶适宜条件筛选 酶解温度、pH、酶量(酶水平)、底物浓度和酶解时间是影响蛋白酶酶解效果的主要因素。为了筛选出适宜的酶解条件，本试验采用单因素试验设计，以氮溶指数(Nitrogen Solution Index，NSI)、水解度(Degree of Hydrolysis，DH)(包括DH_1和DH_2)和苦味值为标识，按酶解温度(T，℃)、pH、酶量([E]/[S]，w/w)、底物浓度([S]，w/v)和酶解时间(t，min)逐一递进的方式分别对碱性蛋白酶(Alcalase，AL)、酸性蛋白酶(Acidic Protease，AP)、风味酶(Flavourzyme，FL)、As．1398中性蛋白酶(As．1398 NeutalProtease，NP)、木瓜蛋白酶(Papain，PA)、胃蛋白酶(Pepsin，PE)和胰蛋白酶(Trypsin，TR)酶解鸡蛋清蛋白的适宜酶解条件进行筛选。结果表明： (1)酶解温度、pH、酶量、底物浓度和酶解时间均明显影响所试7种蛋白酶酶解鸡蛋清蛋白的效果； (2)所试7种蛋白酶酶解鸡蛋清蛋白的效果存在明显差异，以FL为最优，而AP和NP较差。 试验二 10种双酶组合适宜条件筛选 在试验一基础上，采用L_9(3~4)正交试验设计，以DH(DH_1)和苦味值为标识，在固定试验一筛选出的适宜底物浓度(均为4％)、酶解温度和pH的条件下，筛选基于畜禽(人)消化道前后段的pH有差异而设计的5种“PE+X”和5种“AP+X”共10种双酶组合中(X代表AL、FL、NP、PA或TR)，3个因素(酶解时间、PE或AP酶量和AL等其它5种蛋白酶酶量)各自3个水平的不同组合酶解鸡蛋清蛋白时的适宜条件。结果表明： (1)酶解时间及PE或AP的酶量是影响10种双酶组合酶解鸡蛋清蛋白效果的主要因素。5种“PE+X”组合的酶解效果普遍优于相应的5种“AP+X”组合，且10种双酶组合的酶解效果以“PE+FL”最优，而“AP+NP”和“AP+TR”较差。 (2)以DH和苦味值为标识，10种双酶组合中多数双酶组合的酶解效果并不优
[Abstract]:In order to explore whether the mixture of small (oligopeptides) produced by enzymatic hydrolysis of egg white protein has immunomodulatory effect and the mechanism of regulating immune function, the in vitro immunity of different mixture of small (oligopeptides) of egg white to healthy mice was investigated on the basis of screening suitable enzymatic hydrolysis conditions and preparing the mixture of small (oligopeptides) produced by enzymatic hydrolysis of egg white. The effects of oligopeptides on the immune function of mice with normal immune function and low immunity were studied. The mechanism of the mixture of oligopeptides and egg white on the immune function was explored.
Part one: screening conditions for protease hydrolysis
Screening of suitable conditions for 7 proteases in Experiment 1
Enzymatic hydrolysis temperature, pH, enzyme content (enzyme level), substrate concentration and enzymatic hydrolysis time were the main factors affecting the enzymatic hydrolysis of protease. In order to select the suitable enzymatic hydrolysis conditions, the single factor experiment design was adopted, with Nitrogen Solution Index (NSI), Degree of Hydrolysis (DH_1 and DH_2) and bitter value as the standard. Recognition, according to the enzymatic hydrolysis temperature (T,, C), pH, enzyme amount ([E]/[S], w/w), substrate concentration ([S], w/v) and enzymatic hydrolysis time (t, min) progressive manner of alkaline protease (AL), acidic protease (AP), flavor enzyme (FL), as. 1398 neutral protease (As. 1398 NeutalProtease, NP), papain (PA), pepsin (T, min), respectively. The optimum conditions for enzymatic hydrolysis of egg white protein by protease (Pepsin, PE) and trypsin (TR) were screened.
(1) The enzymatic hydrolysis temperature, pH, enzyme amount, substrate concentration and enzymatic hydrolysis time significantly affected the enzymatic hydrolysis effect of the seven proteases.
(2) The results of enzymatic hydrolysis of egg albumin by seven proteases showed significant difference. FL was the best, but AP and NP were poor.
Experiment two selection of 10 suitable conditions for two enzyme combinations
On the basis of experiment 1, 5 PE+X and 5 AP+X combinations were screened by L_9 (3~4) orthogonal design, marked by DH (DH_1) and bitter value, under the conditions of suitable substrate concentration (all 4%), enzymatic hydrolysis temperature and pH value, which were selected from fixed experiment 1. The optimum conditions for enzymatic hydrolysis of egg white protein by different combinations of three levels (X for AL, FL, NP, PA or TR), three factors (enzymatic hydrolysis time, PE or AP enzyme amount, and AL and other five proteinases amount) were determined.
(1) The enzymatic hydrolysis time and the enzyme content of PE or AP were the main factors affecting the enzymatic hydrolysis of egg white protein by 10 combinations of double enzymes. The enzymatic hydrolysis effect of 5 combinations of PE+X was generally better than that of 5 combinations of AP+X, and the enzymatic hydrolysis effect of 10 combinations of PE+FL was the best, while that of AP+NP and AP+TR was the worst.
(2) with DH and bitterness value as identification, most of the two enzyme combinations in the 10 double enzyme combinations did not achieve satisfactory results.
【学位授予单位】：四川农业大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R392
本文编号：2193283
[Abstract]:In order to explore whether the mixture of small (oligopeptides) produced by enzymatic hydrolysis of egg white protein has immunomodulatory effect and the mechanism of regulating immune function, the in vitro immunity of different mixture of small (oligopeptides) of egg white to healthy mice was investigated on the basis of screening suitable enzymatic hydrolysis conditions and preparing the mixture of small (oligopeptides) produced by enzymatic hydrolysis of egg white. The effects of oligopeptides on the immune function of mice with normal immune function and low immunity were studied. The mechanism of the mixture of oligopeptides and egg white on the immune function was explored.
Part one: screening conditions for protease hydrolysis
Screening of suitable conditions for 7 proteases in Experiment 1
Enzymatic hydrolysis temperature, pH, enzyme content (enzyme level), substrate concentration and enzymatic hydrolysis time were the main factors affecting the enzymatic hydrolysis of protease. In order to select the suitable enzymatic hydrolysis conditions, the single factor experiment design was adopted, with Nitrogen Solution Index (NSI), Degree of Hydrolysis (DH_1 and DH_2) and bitter value as the standard. Recognition, according to the enzymatic hydrolysis temperature (T,, C), pH, enzyme amount ([E]/[S], w/w), substrate concentration ([S], w/v) and enzymatic hydrolysis time (t, min) progressive manner of alkaline protease (AL), acidic protease (AP), flavor enzyme (FL), as. 1398 neutral protease (As. 1398 NeutalProtease, NP), papain (PA), pepsin (T, min), respectively. The optimum conditions for enzymatic hydrolysis of egg white protein by protease (Pepsin, PE) and trypsin (TR) were screened.
(1) The enzymatic hydrolysis temperature, pH, enzyme amount, substrate concentration and enzymatic hydrolysis time significantly affected the enzymatic hydrolysis effect of the seven proteases.
(2) The results of enzymatic hydrolysis of egg albumin by seven proteases showed significant difference. FL was the best, but AP and NP were poor.
Experiment two selection of 10 suitable conditions for two enzyme combinations
On the basis of experiment 1, 5 PE+X and 5 AP+X combinations were screened by L_9 (3~4) orthogonal design, marked by DH (DH_1) and bitter value, under the conditions of suitable substrate concentration (all 4%), enzymatic hydrolysis temperature and pH value, which were selected from fixed experiment 1. The optimum conditions for enzymatic hydrolysis of egg white protein by different combinations of three levels (X for AL, FL, NP, PA or TR), three factors (enzymatic hydrolysis time, PE or AP enzyme amount, and AL and other five proteinases amount) were determined.
(1) The enzymatic hydrolysis time and the enzyme content of PE or AP were the main factors affecting the enzymatic hydrolysis of egg white protein by 10 combinations of double enzymes. The enzymatic hydrolysis effect of 5 combinations of PE+X was generally better than that of 5 combinations of AP+X, and the enzymatic hydrolysis effect of 10 combinations of PE+FL was the best, while that of AP+NP and AP+TR was the worst.
(2) with DH and bitterness value as identification, most of the two enzyme combinations in the 10 double enzyme combinations did not achieve satisfactory results.
【学位授予单位】：四川农业大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R392
【引证文献】
相关期刊论文 前2条
1 韩樱;何慧;马芝丽;张文君;李江涛;;蛋清肽及肽钙配合物的制备[J];食品科学;2011年06期
2 牛慧慧;马美湖;杨昆;;蛋清肽的抗氧化稳定性与功能特性[J];食品科学;2011年15期
相关硕士学位论文 前1条
1 纪丽娜;金枪鱼头酶解物免疫活性肽的分离及对小鼠腹腔巨噬细胞功能的影响[D];广东海洋大学;2012年
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