脱脂蛋黄蛋白粉酶水解实验研究

蛋黄蛋白粉是提取药用卵磷脂生产的副产物,用酶解方法将蛋黄蛋白转化为氨基酸或寡肽是回收利用脱脂蛋黄蛋白粉的重要途径之一.从研究水解反应入手,通过单因素试验和正交优化试验研究胃蛋白酶的最佳水解条件,考察了底物浓度、水解时间、反应温度、pH、酶浓度对脱脂蛋黄蛋白粉水解度的影响,然后根据正交试验结果对胃蛋白酶胰蛋白酶组合水解进行研究.研究结果表明,胃蛋白酶水解的最佳工艺条件是:温度为38 ℃,pH为2.0,底物浓度为5％,酶浓度为2％,水解时间为4h,此条件下水解度为15.76％.胃蛋白酶-胰蛋白酶组合酶水解的最佳工艺条件是:先加胃蛋白酶水解4h,而后调节pH为7.5,温度55℃,再加入浓度为0.5％的胰蛋白酶水解4h,此时水解度高达20.41％,明显高于胃蛋白酶单酶水解.     

酶解猪血浆蛋白粉制备ACE抑制肽的工艺优化

选用胃蛋白酶、胰蛋白酶、碱性蛋白酶和中性蛋白酶水解猪血浆蛋白粉,通过单因素和正交试验考察其水解产物对ACE的抑制活性,并优选酶解工艺条件。结果显示:胃蛋白酶适宜水解血浆蛋白粉,制备ACE抑制肽;影响胃蛋白酶水解的4个因素主次顺序为底物质量浓度>酶解时间>pH值>酶与底物质量比(m酶:m底物),其中底物质量浓度和酶解时间的影响显著(P<0.05),pH值和m酶:m底物的影响不显著(P>0.05);适宜胃蛋白酶水解的条件为pH2.3、水解时间1.5h、底物质量浓度1g/100mL、m酶:m底物1:6。     

酶法制取土鳖虫氨基酸口服液的研究

采用酶法水解土鳖虫制取氨基酸口服液。其中胰蛋白酶对土鳖虫蛋白的水解能力最强。最佳水解条件为：pH8.0，温度45℃，底物浓度7．5％，酶与底物比6000μ／g水解时间6h。进一步研究发现，采用胃蛋白酶同胰蛋白酶联合水解，其氨基氮生成率比用单一的酶高。水解液中含有18种氨基酸，8种必需氨基酸齐全。氮溶指数达90％，是一种营养价值较高的蛋白质补剂。     

双酶水解乳清蛋白ACE抑制肽的制备工艺优化

以乳清蛋白为原料,采用胃蛋白酶和胰蛋白酶双酶先后水解乳清蛋白,通过单因素试验和正交试验优化胃蛋白酶和胰蛋白酶水解乳清蛋白制备血管紧张素转换酶(ACE)抑制肽的工艺,将水解物以3 kDa超滤膜过滤,研究表明,胃蛋白酶水解乳清蛋白最佳酶解工艺条件为水解温度37℃、底物质量浓度6 g/100 mL、酶与底物比3 728 U/g,此时乳清蛋白ACE抑制率为86%;胰蛋白酶水解乳清蛋白最佳酶解条件为温度55℃、底物质量浓度6 g/100 mL、酶与底物比3 480 U/g,此时ACE抑制率为72%。利用超滤离心管获得分子量小于3 kDa的乳清蛋白ACE抑制率96%。     

消化道酶连续水解制备苦荞球蛋白多肽及其抗氧化活性研究

采用胃蛋白酶、胰蛋白酶对苦荞球蛋白进行连续水解,以水解度(DH%)为指标,采用单因素分析底物浓度、酶底比、pH值、水解时间对苦荞球蛋白的酶解效果,并通过正交实验对胃-胰蛋白酶连续作用的酶解工艺进行优化;检测酶解产物的体外抗氧化活性。结果表明:苦荞球蛋白经胃蛋白酶酶解的最佳条件为:底物浓度30 g/L、酶底比为10%、pH值为2.0、反应时间为2.0 h;胰蛋白酶酶解的最佳条件为:底物浓度30 g/L、酶底比为5%、pH值为8、反应时间为2.5 h;胃-胰蛋白酶连续酶解最佳条件:经胃蛋白酶先水解1 h后再继续用胰蛋白酶水解1.5h,水解度为25.18%。和苦荞球蛋白相比,苦荞球蛋白酶解物的抗超氧阴离子能力及羟自由基清除能力均增强(p0.05)。     

双酶水解螺旋藻藻胆蛋白制备ACE抑制肽的工艺优化

以螺旋藻为原料,采用反复冻融和超声波解冻结合法提取藻胆蛋白,利用等电点分离藻胆蛋白,采用SDS-PAGE电泳确定分离蛋白的种类。采用胃蛋白酶和胰蛋白酶双酶先后水解藻胆蛋白。通过单因素实验和正交实验优化胃蛋白酶和胰蛋白酶水解藻胆蛋白制备血管紧张素转换酶(ACE)抑制肽的工艺,研究表明:胃蛋白酶水解藻胆蛋白最佳酶解工艺条件是水解温度为37℃,底物质量浓度为6%(w/v),酶与底物比为5220 U/g,此时ACE抑制率为82.07%。胰蛋白酶水解藻胆蛋白最佳酶解条件是温度为42℃,底物质量浓度为6%(w/v),酶与底物比为5220 U/g,此时ACE抑制率为80.35%。利用超滤离心管获得分子量小于3 kDa的藻胆蛋白ACE抑制率94.30%。     

胃蛋白酶胰蛋白酶水解酪蛋白的研究

主要研究胃蛋白酶、胰蛋白酶水解酪蛋白的过程.首先通过正交试验研究了胃蛋白和胰蛋白酶水解酪蛋白的最佳条件,分别为酶浓度0.15%、温度55℃、pH7.5、底物浓度6%和酶浓度1%、温度55℃、pH=1.6、底物浓度1%.然后对于胃蛋白酶胰蛋白酶组合水解进行研究,最后通过层析分离和液相色谱初步验证了水解产物含有β-啡肽-7等生物活性多肤.     

林蛙油蛋白水解营养液的研究

目的 研究林蛙油蛋白水解营养液的工艺条件.方法 采用胰蛋白酶和木瓜蛋白酶分步水解林蛙油,酶解液中添加β-环状糊精、柠檬酸、蜂蜜调配.然后均质、装罐、杀菌.结果 林蛙油蛋白水解营养液配方为β-环状糊精添加量2.0％,柠檬酸添加量0.15％,蜂蜜添加量8％.杀菌条件105℃,20 min.结论 林蛙油蛋白水解营养液营养丰富...     

林蛙油蛋白水解营养液的研究

目的研究林蛙油蛋白水解营养液的工艺条件。方法采用胰蛋白酶和木瓜蛋白酶分步水解林蛙油,酶解液中添加β-环状糊精、柠檬酸、蜂蜜调配,然后均质、装罐、杀菌。结果林蛙油蛋白水解营养液配方为β-环状糊精添加量2.0%,柠檬酸添加量0.15%,蜂蜜添加量8%。杀菌条件105℃,20 min。结论林蛙油蛋白水解营养液营养丰富,口感、风味均佳。     

木瓜蛋白酶作用于林蛙油蛋白质水解功能特性的研究

本研究以林蛙油为底物,加酶水解,实验确定林蛙油水解最适宜的酶制剂为木瓜蛋白酶。与林蛙油相比,水解产物具有很好的溶解性,游离雌二醇含量随水解程度的增加而增加,粘度随水解程度的增加而降低。     

不同酶切方式引发大豆蛋白构象变化及功能特性评价

本研究以枯草杆菌碱性蛋白酶、菠萝蛋白酶、枯草杆菌中性蛋白酶、胃蛋白酶和胰蛋白酶酶解大豆分离蛋白产物为研究对象,借助十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、紫外吸收光谱、荧光光谱、红外光谱解析不同酶切条件下蛋白产物构象变化,并对其乳化性及抗氧化活性等功能特性进行评价。结果表明：酶解促进蛋白解折叠,内部氨基酸暴露,并受到酶切方式的影响;菠萝蛋白酶酶解作用范围宽泛,疏水性氨基酸浓度达到最大值(27.10±0.08)%,氨基酸存在微环境发生改变,形成柔性结构单元,其乳化性及抗氧化性都得到明显改善,为具有最佳功能特性的酶解蛋白;酶特异性作用位点导致酶解作用程度较低的枯草杆菌碱性蛋白酶、枯草杆菌中性蛋白酶、胃蛋白酶及胰蛋白酶水解产物功能特性均高于未处理样品。值得注意的是,水解程度最低的胰蛋白酶水解产物呈现优异乳化稳定性,这是因为大豆中胰蛋白酶抑制剂抑制蛋白过度水解,提升蛋白溶解度同时,肽段长度适宜稳定包裹油滴。     

条斑紫菜活性肽的制备及其功能性质研究

采用木瓜蛋白酶、胰蛋白酶、胃蛋白酶水解紫菜蛋白制备活性肽,以水解度为评价指标,优化酶水解条件;研究条斑紫菜活性肽的溶解性、乳化性与乳化稳定性、起泡性与泡沫稳定性等功能性质.结果表明,3种酶解物都具有良好的溶解性且在等电点附近不沉淀,乳化性及其稳定性不如紫菜蛋白且随着pH升高而增强,酸性条件下起泡性较好,胰蛋白酶解物起泡性最强.     

龙须菜蛋白的提取工艺优化及降血压组分制备

目的 优化龙须菜蛋白的提取工艺,并制备降血压组分.方法 从胃蛋白酶、中性蛋白酶、碱性蛋白酶和胰蛋白酶中筛选能获得最佳蛋白提取率的蛋白酶,采用单因素试验考查pH、底物浓度、酶解温度、酶底比、酶解时间对蛋白提取率和ACE抑制率的影响,采用响应面法确定最佳工艺条件,采用超滤膜分离技术龙须菜蛋白酶解液中制备血管紧张素转换酶(a...     

Functional properties of limited hydrolysed cross‐linked casein–gelatin composites

A casein–gelatin composite was prepared by cross‐linking of caseinate and bovine gelatin with a microbial transglutaminase, and the impact of limited proteolysis by trypsin on some functional properties of the composite was investigated in the present work. Two hydrolysed composites were prepared with degree of hydrolysis (DH) of 1 and 2% and analysed by SDS‐PAGE to reflect polypeptide profiles. Some functional properties of the two hydrolysed composites were evaluated, among which solubility, digestibility in vitro, surface hydrophobicity and emulsifying properties showed dependence on the DH. Limited proteolysis of the composite improved its solubility in pH 3–10, especially when the DH was 2%. Compared to the composite, the hydrolysed composites showed an increased emulsifying activity index (about 357–408%), emulsion stability index (about 23–28%) and digestibility in vitro (about 65–80% for pepsin, whereas 2–3% for pepsin–trypsin hydrolysis), together with a decreased surface hydrophobicity (about 55–62%) and oil absorption capacity (about 20–23%). The applied proteolysis also led to the aqueous dispersions of the two hydrolysed composites much lower apparent viscosity, storage and loss modulus.     

Effect of Partial Proteolysis and Succinylation on Functionality of Corn Germ Protein Isolate

Corn germ protein isolate (CGPI) was partially hydrolyzed with trypsin and pepsin and succinylated at three levels. Various functional and electrophoretic properties of the native and modified protein were determined. Water absorption and foaming properties of CGPI were Improved by partial hydrolysis with trypsin; emulsifying capacity and nitrogen solubility were reduced; oil absorption was increased only slightly. CGPI pepsin hydrolyzate has decreased oil absorption, nitrogen solubility and emulsifying capacity but improved foaming properties; water absorption was unchanged. Treatment of CGPI with succinic anhydride improved water and oil absorption, nitrogen solubility and foaming capacity but decreased emulsifying capacity; foam stability was unchanged. Succinylation retarded electropohoretic mobility while hydrolysis altered band intensities.     

辅酶Q10软胶囊的急性毒性和遗传毒性研究

目的:研究辅酶Q10软胶囊的急性毒性和遗传毒性,为辅酶Q10的进一步开发利用提供安全依据。方法:应用小鼠急性经口毒性试验、Ames试验、小鼠骨髓嗜多染红细胞微核试验和小鼠精子畸形试验对某种辅酶Q10软胶囊的毒性进行试验观察。结果:该辅酶Q10软胶囊对雌雄小鼠LD50均大于10 000mg/kg.bw,属实际无毒物质;Ames试验中受试物各剂量组回变菌落数均未超过自发回变菌落数的2倍,亦无剂量-反应关系,结果为阴性;小鼠骨髓嗜多染红细胞微核试验和小鼠精子畸形试验中各剂量组微核率、精子畸形率与阴性对照组之间无显著性差异(P0.05),而环磷酰胺阳性对照组与阴性对照组之间有极显著性差异(P0.01),证明试验结果均为阴性。结论:本次试验条件下,该辅酶Q10软胶囊属实际无毒级,未显示有急性毒性和遗传毒性作用。     

限制性酶解对豆粉相关性质的影响

为降低豆粉的致敏性,扩大其应用,采用限制性酶解制备豆粉。以木瓜蛋白酶、风味蛋白酶和碱性蛋白酶为限制性酶解用酶,研究不同的酶制剂及酶解时间对豆粉的致敏性、溶解性、表面疏水性及乳化性的影响,并对豆粉进行感官评价。结果表明:随着酶解时间的延长,豆粉的致敏性降低,溶解性、表面疏水性、乳化活性和乳化稳定性增加;其中利用木瓜蛋白酶酶解30 min制备的豆粉致敏原含量最低,为1. 95%,溶解性最高,为88. 55%,乳化活性及乳化稳定性最大,分别为182. 4 m2/g和120. 8 min;利用SDS-PAGE电泳发现,酶解作用使豆粉蛋白质中的7S和11S降解,生成小分子的肽类,从而降低豆粉的致敏性;利用木瓜蛋白酶酶解20 min制备的豆粉口感与传统市售豆粉相似。     

酶解制备猪小肠粘膜蛋白粉工艺条件优化

为研究蛋白酶酶解制备猪肠粘膜蛋白粉工艺,以天然猪肠衣加工副产物肠粘膜为原料,以酶解产物中可溶性蛋白含量为指标,通过单因素实验和响应面实验,筛选适宜用于酶解猪小肠粘膜的蛋白酶并对其酶解工艺条件进行优化。结果表明,木瓜蛋白酶酶解猪肠粘膜制备蛋白粉可溶性蛋白含量达7.37%±0.06%,显著高于胰蛋白酶和胃蛋白酶酶解制备得到的肠粘膜蛋白粉可溶性蛋白含量,最佳酶解工艺条件为:水解时间6.5 h,酶添加量5400 U/g蛋白,液固比4 mL/g,在此条件下酶解产物中可溶性蛋白含量为15.81%±0.04%,这表明酶法制备肠粘膜蛋白粉工艺合理可行,为动物副产物的开发利用提供参考依据。     

Peptic and tryptic hydrolysis of native and heated whey protein to reduce its antigenicity

This study examined the effects of enzymes on the production and antigenicity of native and heated whey protein concentrate (WPC) hydrolysates. Native and heated (10 min at 100°C) WPC (2% protein solution) were incubated at 50°C for 30, 60, 90, and 120 min with 0.1, 0.5, and 1% pepsin and then with 0.1, 0.5, and 1% trypsin on a protein-equivalent basis. A greater degree of hydrolysis was achieved and greater nonprotein nitrogen concentrations were obtained in heated WPC than in native WPC at all incubation times. Hydrolysis of WPC was increased with an increasing level of enzymes and higher incubation times. The highest hydrolysis (25.23%) was observed in heated WPC incubated with 1% pepsin and then with 1% trypsin for 120 min. High molecular weight bands, such as BSA, were completely eliminated from sodium dodecyl sulfate-PAGE of both native and heated WPC hydrolysates produced with pepsin for the 30-min incubation. The α-lactalbumin in native WPC was slightly degraded when incubated with 0.1% pepsin and then with 0.1% trypsin; however, it was almost completely hydrolyzed within 60 min of incubation with 0.5% pepsin and then with 0.5% trypsin. Incubation of native WPC with 1% pepsin and then with 1% trypsin for 30 min completely removed the BSA and α-lactalbumin. The β-lactoglobulin in native WPC was not affected by the pepsin and trypsin treatments. The β-lactoglobulin in heated WPC was partially hydrolyzed by the 0.1 and 0.5% pepsin and trypsin treatments and was completely degraded by the 1% pepsin and trypsin treatment. Antigenicity reversibly mimicked the hydrolysis of WPC and the removal of β-lactoglobulin from hydrolysates. Antigenicity in heated and native WPC was reduced with an increasing level of enzymes. A low antigenic response was observed in heated WPC compared with native WPC. The lowest antigenicity was observed when heated WPC was incubated with 1% pepsin and then with 1% trypsin. These results suggested that incubation of heated WPC with 1% pepsin and then with 1% trypsin was the most effective for producing low-antigenic hydrolysates by WPC hydrolysis and obtaining low molecular weight small peptides. Further research is warranted to identify the low molecular weight small peptides in the WPC hydrolysates produced by pepsin and trypsin, which may enhance the use of whey.     

胰蛋白酶限制性水解脱酚棉籽蛋白的研究

采用胰蛋白酶对脱酚棉籽蛋白进行限制性酶解,以改善棉籽蛋白在中性条件下的溶解性.以酶解棉籽蛋白水解度和溶解度为指标,对胰蛋白酶水解脱酚棉籽蛋白的条件进行了优化.结果表明,胰蛋白酶水解脱酚棉籽蛋白的优化条件为:酶底比2∶ 1(50 000 U/g),底物质量浓度100 g/L,pH 10,于20 ℃下酶解3 h.在此条件下,酶解棉籽蛋白的水解度为10.87%,溶解度为63.48%.与酶解前相比,溶解度提高了43.82%.溶解度分析结果表明,限制性酶解棉籽蛋白在pH 1～11范围内均具有较好的溶解度,仍然保持了棉籽蛋白酸溶性的特点.