Alcalase 2.4L碱性内切酶和Flavourzyme风味酶酶解大豆分离蛋白及脱苦工艺优化

以大豆分离蛋白为原料,选用Alcalase 2.4L碱性内切酶和Flavourzyme风味蛋白酶对大豆分离蛋白进行酶法水解及脱苦工艺研究。以水解度和苦味分值为考察值,对酶解工艺进行优化,确定最佳条件。结果表明:Alcalase2.4L碱性内切酶最佳酶解条件为加酶量14 000 U/g、酶解温度60℃、酶解pH8.5、底物质量分数5%,酶解时间2h,最终水解度为45.34%,此时水解液苦味值为4。Flavourzyme风味蛋白酶对水解液进行二次水解的最优酶解条件为加酶量300 U/g、酶解温度55℃、酶解pH 7.0、酶解时间3 h,此条件下大豆分离蛋白水解液苦味值最低为1.2。Alcalase2.4L碱性内切酶和Flavourzyme风味蛋白酶水解大豆分离蛋白使水解度得到较大提高的同时也解决了水解液的苦味问题。     

复合酶水解猪血液工艺条件的研究

为了探索出经济实用的酶解猪血的方法,对两种不同来源的蛋白酶其采用单一和复合酶水解鲜猪血的酶解效果进行了比较。结果表明:复合法酶解鲜猪血水解效果最好。复合酶最适水解工艺条件为最初pH7～8、温度50℃、酶底物浓度比(E/S)为6000U/g、底物浓度8%、水解时间8h。     

基于人工神经网络的牡蛎蛋白酶解动力模型的构建

以牡蛎分离蛋白为底物,采用碱性蛋白酶(Alcalase)进行酶解,在获得一定实验数据基础上利用训练后的人工神经网络(ANN)模型对牡蛎分离蛋白酶解过程进行预测。结果表明:训练后的ANN模型决定系数R2达到0.9998,人工神经网络预测水解度DH值和实验DH值之间具有很强的相关性,相关系数r值达到0.9957。并且在一定的酶浓度及底物浓度范围内,采用ANN预测数据,双倒数作图所得回归方程决定系数R2值达到0.9740,计算得米氏常数Km和最大反应速度V max分别为26.1g/L,8.6g/(L·min)。研究结果为人工神经网络模型在食品蛋白酶促反应动力学方面的应用提供参考。      

超声和Alcalase 酶复合处理水解大豆分离蛋白工艺研究

以大豆分离蛋白为底物,通过单因素试验和正交试验,确定超声和Alcalase 酶复合处理对大豆分离蛋白水解的最佳条件。结果表明,最佳水解条件为大豆分离蛋白质量分数5.0%、超声处理时间30min、加酶量5.0%、酶解pH8.0、酶解温度55℃、酶解时间4.0h,在此条件下,大豆分离蛋白水解度为12.21%。     

碱性蛋白酶控制酶解诱导大豆分离蛋白纳米颗粒的形成机制

以大豆分离蛋白（soy protein isolate,SPI）为原料,利用碱性蛋白酶对其进行酶解处理（0～24 h）,探究SPI的结构变化规律,发现碱性蛋白酶控制酶解可诱导SPI自组装形成系列分布均匀（多相分散系数＜0.3）、粒径可控（90～200 nm）且具有不同表面特性的大豆蛋白纳米颗粒（soy protein nanoparticles,SPNs）,其中水解度（degree of hydrolysis,DH）及亚基解离/降解是影响SPNs形成的关键性因素。酶解初期（10～30 min,DH约3%）,SPI中β-伴大豆球蛋白（7S）组分α与α’亚基的部分降解有利于两亲性结构的释放,提高蛋白表面疏水性,降低临界聚集浓度,形成包含相对完整的7S及大豆球蛋白（11S）亚基的I类纳米颗粒（SPNs-DH 3%）。随着酶解时间的延长（1～2 h）,α与α’亚基的进一步降解促进了疏水性β亚基与B亚基的暴露,增强的疏水相互作用导致体系浊度增加,其中可溶性聚集体向不溶性疏水聚集的转化使得蛋白表面疏水性急剧下降,形成以A亚基及部分β亚基为主导的II类亲水型纳米颗粒（SPNs-DH 5%）。酶解后期（4～24 h）,A亚基的进一步降解则产生更多亲水性多肽,不利于纳米颗粒的形成。进而探究SPNs的形成机制,圆二色光谱结构表明,SPNs的形成与蛋白α-螺旋和无规卷曲结构向β-折叠转化有关。两类SPNs的整体结构均由疏水相互作用维持,而氢键和二硫键分别参与颗粒表面与内部结构的形成。与SPNs-DH 3%相比,SPNs-DH 5%中形成了更多由二硫键与氢键稳定的折叠结构。此外,由于酶解过程中不断释放抗氧化肽段,其所形成SPNs的抗氧化性较原始SPI均有所提升。     

大豆分离蛋白的酶法水解研究

本试验主要从预处理温度、预处理时间、底物浓度、加酶量、酶解温度、酶解时间等方面系统研究了AS1·398中性蛋白酶对大豆分离蛋白水解的影响,并运用正交试验设计和方差分析得到水解最佳条件。     

Effects of limited enzymatic hydrolysis with trypsin on the functional properties of hemp (Cannabis sativa L.) protein isolate

Effects of limited enzymatic hydrolysis induced by trypsin on the physicochemical and functional properties of hemp (Cannabis sativa L.) protein isolate (HPI) were investigated. The enzymatic hydrolysis was confirmed by sodium dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and size exclusion chromatography (SEC). SEC and differential scanning calorimetry (DSC) analyses confirmed the presence of aggregates in the corresponding hydrolysates (with the degree of hydrolysis of 2.3–6.7%). Functional properties, including protein solubility (PS), thermal properties, emulsifying and foaming properties, and water holding and fat adsorption capacities (WHC and FAC) were evaluated. The PS was remarkably improved by the limited enzymatic hydrolysis at all tested pH values. However, the enzymatic hydrolysis led to the marked decreases in emulsifying activity index, foaming capacity and foam stability, WHC and FAC. These decreases were to a great extent related to the presence of aggregates in the hydrolysates.     

Characterisation and functional properties of proteins of some Indian chickpea (Cicer arietinum) cultivars

BACKGROUND: Chickpea (Cicer arietinum L.) proteins have received attention during recent years owing to their higher biological values and better functional ingredients than oilseed proteins. In this study the composition, fractionation, electrophoretic behaviour and functional properties of five chickpea protein concentrates were determined. RESULTS: The chickpea proteins contained 15.9–54.8 g kg^?1 albumin, 48.9–154.1 g kg^?1 globulin, 39.2–76.5 g kg^?1 glutelin and traces of prolamin. Electrophoresis of the various fractions revealed that albumin and globulin were made up of sub‐units of different molecular weights ranging from 7 to 96 kDa. Water and oil absorption of the protein concentrates varied from 1.15 to 2.75 g g^?1 and from 2.60 to 5.65 g g^?1 respectively. Foaming capacity and foam stability of the protein concentrates were good and improved with the addition of salt (10 g L^?1 NaCl) or sugar (100 g L^?1 sucrose) at both isoelectric and neutral pH. Emulsifying capacity and emulsion stability of the protein concentrates were good and excellent respectively. CONCLUSION: Protein concentrates prepared from chickpeas have potential use in food formulations owing to their good emulsifying/foaming and water/oil‐binding capacities. Copyright © 2008 Society of Chemical Industry     

核桃蛋白酶法水解工艺条件研究

研究了核桃蛋白酶法水解的工艺条件,结果表明:蛋白酶种类对核桃蛋白水解作用影响较大,Alcalase 2.4L、Neutrase 0.8L对核桃蛋白水解作用较强;Alcalase 2.4L较适宜的酶解条件为酶与底物浓度比1000U/g,pH 8.0,温度60℃;Neutrase 0.8L较适合的水解条件为酶浓度为2000U/g,pH 6.0,温度45℃;Alcalase 2.4L、Neutrase 0.8L复合酶可以对核桃蛋白进行连续水解,并能提高核桃蛋白的水解度,产物肽链长度趋近于5。     

不同酶切方式对乳清蛋白疏水性和乳化性的影响

采用不同的蛋白酶对乳清蛋白进行水解,考察了肽键断裂方式对乳清蛋白肽疏水性和乳化性的影响,以及乳清蛋白不同酶解产物的疏水性和乳化性的关系。结果表明:不同蛋白酶作用于乳清蛋白得到的水解产物疏水性不同,6种蛋白酶解液的疏水性均随水解度的增大而降低,其中以胰凝乳蛋白酶酶解液的疏水性下降的最慢。研究还发现,乳化性随着水解度增加而先升高后下降,以双酶复合酶解液最差。不同蛋白酶水解液的乳化性指数随疏水性指数的降低而升高,乳化性指数与疏水性氨基酸质量分数成正相关。     

Alcalase对大豆分离蛋白凝胶性质的影响

研究了Alcalase蛋白酶对大豆分离蛋白凝胶形成过程中温度、酶的添加量、酶反应速率及水解度对凝胶体系流变学性质的影响及蛋白质各亚基在水解过程中的变化情况。结果表明：反应存在着温度限制，同时也受水解度和酶添加量的影响。在较低温度：20℃、30℃时能得到较高的凝胶强度；温度升高，凝胶强度减弱。低温下，较大的酶添加量有利于反应体系的胶凝，而高温下，较低的酶添加量才有利于体系的胶凝。低的水解度下有利于形成稳定的凝胶，40℃时水解度超过8％就不能形成稳定的凝胶。经Alcalase作用后，大豆分离蛋白的7S和11S球蛋白均有不同程度的水解。     

Enzymatic hydrolysis and their effects on conformational and functional properties of peanut protein isolate

The effects of limited enzymatic hydrolysis by Alcalase on the conformational and functional properties of peanut (Arachis hypogaea L.) protein isolate (PPI) were investigated. Acid subunits of arachin were most susceptible to Alcalase hydrolysis, followed by conarachin and the basic subunits of arachin. Enzymatic hydrolysis increased the thermal stability of arachin and led to a sharp increase in the number of disulphide bonds with a decrease of the sulphydryl group in PPI hydrolysates in comparison with PPI. The analysis of intrinsic fluorescence spectra indicated a more moveable tertiary conformation of PPI hydrolysates than PPI. The limited emzymatic hydrolysis improved the functional properties of PPI, such as protein solubility and gel-forming ability, but impaired the emulsifying activity index.     

具有金属螯合活性的蚕豆蛋白酶解物的制备

目的探究蚕豆蛋白酶解物的金属螯合活性,研究其金属螯合活性与其抗氧化活性的关系。方法分别采用碱性蛋白酶、中性蛋白酶、木瓜蛋白酶对蚕豆蛋白进行酶解,并测定其水解物的抗氧化活性与金属螯合活性,选用碱性蛋白酶为酶解蚕豆蛋白制取金属螯合肽的最适酶,以酶解产物的水解度、抗氧化活性及金属螯合活性为测定指标获得合适的水解条件。结果 3种蛋白酶的蚕豆蛋白酶解产物都有金属螯合活性和抗氧化活性,碱性蛋白酶为酶解蚕豆蛋白的最适酶,最适酶解时间为4 h时,得到的酶解产物金属离子螯合率为88.22%,抑制羟自由基能力为220.70 U/mg,总还原力为0.03 U/mg。结论蚕豆蛋白酶解物具有一定的金属离子螯合活性与抗氧化活性,水解度对蚕豆蛋白酶解物的金属离子螯合活性及抗氧化活性有明显的影响,蚕豆蛋白酶解物的金属螯合活性与总还原力及抑制羟自由基能力呈现显著的正相关性,相关系数分别为0.925、0.968(P0.01)。     

芝麻蛋白制备金属螯合肽的酶解工艺研究

分别采用木瓜蛋白酶、胰蛋白酶和碱性蛋白酶Alcalase对芝麻蛋白进行水解,结果表明胰蛋白酶为制备金属螯合肽的最佳酶。通过优化胰蛋白酶酶解工艺条件发现最佳条件为:底物质量浓度5%(g/100mL),酶添加浓度20u/g(底物),水解时间5h,得到的酶解产物金属螯合率最强,与Fe2+的螯合率为90.9%,与Zn2+的螯合率为93.5%。通过考察水解度对酶解产物金属螯合率及抗氧化能力的影响,结果显示水解度在18.9%到22.4%之间的酶解产物金属螯合率强,且水解度在一定范围内金属螯合率和抗氧化能力均与水解度呈正相关性。     

Enzymatic hydrolysis of hemp (Cannabis sativa L.) protein isolate by various proteases and antioxidant properties of the resulting hydrolysates

Enzymatic hydrolysis of hemp protein isolate (HPI) by six proteases (alcalase, flavourzyme, neutrase, protamex, pepsin and trypsin) and antioxidant activities of the resulting hydrolysates, obtained for 2 and 4 h were investigated. The yield of trichloroacetic acid (TCA)-soluble peptides (Y_sp), protein composition and surface hydrophobicity (H_o) of the hydrolysates were evaluated. The results showed that the hydrolysates exhibited varying DPPH radical scavenging (with lowest IC₅₀, ∼2.3 mg/mL) and Fe²⁺⁺ chelating (with lowest IC₅₀ of 1.6–1.7 mg/mL) abilities and reducing power (with highest absorbance at 700 nm of 0.31–0.35), depending on their Y_sp and H_o values. The DPPH radical scavenging and Fe²⁺⁺ chelating abilities of the hydrolysates were positively correlated with their Y_sp or H_o values. The results suggest that enzymatic hydrolysis can be used as an effective technique to produce high value-added products of hemp proteins.     

微生物发酵法产大豆多肽液水解度的测定

建立了一种适于蛋白质水解液水解度测定的新方法。以脱脂豆粕粉为原料，具产蛋白酶能力的微生物为生产菌种进行发酵。微生物所产蛋白酶作用于豆粕粉中的大豆蛋白将其水解成大豆多肽。通过新建立的方法测定了不同发酵时间发酵液的水解度，并与文献所报道的常规测定方法进行比较。结果表明，新建立的水解度直接测定法快速简便，灵敏度高，可重复性强，不仅适于微生物发酵法产大豆多肽液水解度的测定，还普遍适用于其它蛋白水解液水解度的测定。     

Influence of degree of hydrolysis on functional properties,antioxidant activity and ACE inhibitory activity of peanut protein hydrolysate

Functional properties, antioxidant and Angiotensin-converting enzyme (ACE) inhibitory activities of peanut protein isolate (PPI) and peanut protein hydrolysate (PPH) prepared using Alcalase, at different (10%, 20%, 30% and 40%) degrees of hydrolysis, (DH) were investigated. Hydrolysis (at DH > 10%) significantly (p < 0.05) improved the solubility (>80%) of PPI, especially in the pH range of 4–6. However, PPI showed better emulsifying and foaming properties than PPH (p < 0.05). As DH increased, ferrous ion chelating activity, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging activity and ACE inhibitory activity of PPH increased, while reducing power decreased (p < 0.05). Bleaching of beta-carotene by linoleic acid was suppressed better by PPI and PPH at 10% DH than of PPH at higher DH. Thus, the results reveal that DH affects functional properties, antioxidant and ACE inhibitory activities of peanut protein.     

Isolation, solubility and in vitro hydrolysis of chickpea vicilin-like protein

The chickpea vicilin-like globulin was isolated and chromatographed on Sepharose CL-6B and Sephacryl S-300. The native globulin with a molecular weight of 140 kDa was resolved in Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in seven polypeptide bands in the range of 12.4-67 kDa. The solubility profile of the protein in water and NaCl solutions was typical of a legume globulin. The purified vicilin-like globulin, native and heated, was hydrolyzed by pepsin, trypsin and chymotrypsin. The hydrolysis patterns indicated that the native vicilin-like protein was only partially degraded by the enzymes in comparison with casein. Heating increased its susceptibility to hydrolysis relative to the native form, for all the enzymes. However, the results obtained by the pH-drop method revealed that the in vitro digestibility of the vicilin-like protein was not altered by heating, while 11S-like and total globulins suffered a small increase, indicating that the structural characteristics of storage globulins may be important factors limiting the protein digestion.     

Alcalase降解水不溶性鸡肉蛋白规律及其产物清除DPPH活性研究

研究了Alcalase降解水不溶性鸡肉蛋白的规律及其产物清除DPPH自由基活性。结果表明,随酶解时间延长,可溶性蛋白质、氨基酸含量不断增加,而肽含量则在水解前8h达到最大,随后不断降低;肽的分子量分布趋势总体表现为大分子量的肽逐渐减少,小分子量肽的含量逐渐增多,肽分子量分布图中肽峰值不断向后推移;Alcalase对分子量在3000～15000Da间的肽段降解能力较弱,水解24h后,这部分肽占总肽量的比值仍为89.93%。水不溶性鸡肉蛋白Alcalase酶解产物有明显的清除DPPH活性,其清除DPPH活性在酶解4h时达到最大,随后不断下降;对于同一酶解时间的产物,浓度越大,其清除DPPH活性越强,但其清除DPPH能力并不与酶解液浓度成倍数关系。      

金带细鲹鱼肉蛋白酶解物水解度与其抗氧化性和功能特性间的相互关系

采用胰蛋白酶水解金带细鲹鱼肉蛋白制备不同水解度(10.8%、14.9%、19.0%和21.7%)的酶解产物,研究水解度与酶解产物抗氧化性和功能特性的关系。抗氧化性分析表明,低水解度处理显著增大了酶解物清除羟基自由基的能力、清除DPPH自由基的能力和螯合亚铁离子的能力(p<0.05),而高水解度处理则增强了酶解产物的金属还原力(p<0.05)。功能特性结果表明:随着水解度的增大,酶解产物的溶解度逐渐增大,而乳化活性和乳化稳定性逐渐降低,起泡性和泡沫稳定性也降低。同时,随着p H的增加,酶解物的溶解性和乳化稳定性呈先下降后上升的趋势,而起泡性和泡沫稳定性则随着p H的增加而降低。上述分析表明,适当的酶解处理对改善金带细鲹鱼肉蛋白的功能特性及抗氧化活性具有一定的作用。