蛋白酶处理对脱脂乳感官品质及游离氨基酸的影响

以新鲜脱脂牛乳为原料,采用分光测色仪、电子舌及氨基酸自动分析仪等分析酶解处理对脱脂牛乳感官品质、游离氨基酸含量及组成的影响。结果表明：经风味蛋白酶处理的脱脂牛乳水解度高,达到24.61%;蛋白酶处理会导致其感官性状的改变,与脱脂牛乳相比,3?种酶解产物L*值均显著（P＜0.05）下降,a*（负值）显著上升（P＜0.05）,风味蛋白酶处理对脱脂牛乳色泽影响大于碱性蛋白酶、复合蛋白酶处理;不同酶解产物滋味轮廓之间存在较大差异,与脱脂牛乳相比其甜味值下降显著,且随酶解时间延长,苦味值上升,甜味值衰退,碱性蛋白酶处理的酶解产物以涩味及涩味回味为主,风味蛋白酶的酶解产物以咸、苦味及苦味回味为主,复合蛋白酶的酶解产物以酸味为主,苦涩等味觉较低,电子舌能较好地区分不同酶解物的滋味差异;酶解处理可使脱脂牛乳中的游离氨基酸及必需氨基酸含量显著增加,苦味氨基酸为主要呈味氨基酸。酶解处理及酶解进程会使脱脂牛乳色泽、滋味及游离氨基酸产生变化,其中风味蛋白酶处理产生的影响大于碱性蛋白酶和复合蛋白酶处理。     

猪肉不同蛋白酶解呈味组分及热反应风味物质比较

采用胰蛋白酶、复合蛋白酶、碱性蛋白酶、中性蛋白酶、木瓜蛋白酶等5种内切酶和风味蛋白酶(外切酶)的各自较佳条件制备猪肉酶解液。胰蛋白酶和风味蛋白酶的酶解液味道较好,内切酶的酶解液水解度均较高,而鲜味较弱、苦味较强。呈味游离氨基酸组成上,风味蛋白酶的酶解液中鲜、甜味氨基酸比例最大,其次是胰蛋白酶的酶解液;而木瓜蛋白酶的酶解液中苦味氨基酸比例最大。超滤分离发现,呈鲜味的均为小于3 ku的组分,尤其小于1 ku的组分鲜味较强。木瓜蛋白酶的酶解液中小于5 ku组分均有苦味,中性蛋白酶的酶解液中1~5ku的组分均有苦味,而胰蛋白酶和风味蛋白酶的酶解液未分离出苦味组分。6种酶解液与木糖反应所得热反应产物均有不同程度的炖煮肉香气,固相微萃取/气-质联机分析发现,胰蛋白酶和碱性蛋白酶的酶解液的热反应产物中,含硫化合物、含氮杂环化合物相对含量较高,而其它4种酶解液的热反应产物中,含氧杂环化合物的相对含量较高。     

紫贻贝酶解条件的研究

本试验主要测定了紫贻贝的基本组分，并利用中性蛋白酶酸性蛋白酶和碱性蛋白酶对紫贻贝酶解，制备蛋白水解液，采用凯氏定氮法和甲醛电位滴定法计算酶解液的水解度，结合酶解液的感官评价和水解度筛选最适合酶和确定酶解的最佳工艺条件。主要结论如下：紫贻贝中基本组分是粗蛋白含量为9．5％，粗脂肪的含量为6．2％，灰分的含量为2．9％，水分的含量为83．2％；中性蛋白酶的最佳酶解条件：时间3h，pH值7，温度50℃，酶的添加量0．3％，此条件下水解度为39．43％；酸性蛋白酶的最佳酶解条件：时间2h，pH值4，温度50℃，酶的添加量0．3％，此条件下水解度为39．43％；碱性蛋白酶的最佳酶解条件：时间3h，pH值8，温度45℃，酶的添加量0．4％，此条件下水解度为46．51％；在最佳酶解条件下，碱性蛋白酶对紫贻贝的酶解效果明显的好于其它两种蛋白酶，但经感官评定，碱性蛋白酶酶解液苦味和氨味较重，不符合海鲜调味品的风味要求，中性蛋白酶和酸性蛋白酶酶解液的苦味和氨味比较轻，适合作为紫贻贝水解的外加酶，综上分析，选择中性蛋白酶和酸性蛋白酶作为紫贻贝水解的外加酶。     

苦荞麦蛋白质的酶法水解研究

以苦荞麦蛋白质作为底物,采用碱性蛋白酶(固体)、胰蛋白酶、碱性蛋白酶Alcalase 2.4L、高效水解蛋白酶、中性蛋白酶、木瓜蛋白酶对其进行酶法水解,并对酶解产物的氨基酸组成和相对分子质量进行研究.结果表明,碱性蛋白酶(固体)的酶解程度最高,在酶解2h时DH(水解度)接近20%,其酶解产物的疏水性氨基酸含量较高,大部分肽链的相对分子质量都低于1000Da.     

分步酶解小麦面筋蛋白制备低苦味肽粉的研究

目的 研究分步酶解小麦面筋蛋白(wheat gluten, WG)制备低苦味肽粉的工艺。方法 选用中性蛋白酶、木瓜蛋白酶、胃蛋白酶水解WG至8%水解度,接着用风味蛋白酶对水解产物进行脱苦处理,对不同酶解产物中苦味肽的特性进行系统研究,探究苦味肽含量、氨基酸组成、分子量分布、表面疏水性等指标变化对WG酶解物苦味值的影响,对比风味蛋白酶对不同单酶酶解物的脱苦效果差异,分析风味蛋白酶对WG酶解物脱苦的内在机理,进而确定制备低苦味小麦蛋白肽粉的最佳酶解工艺。结果 中性蛋白酶的酶解产物经风味蛋白酶作用后,脱苦效果最显著,苦味肽苦味值从4.08降至2.25,酶解产物的苦味值可下降56.42%。木瓜蛋白酶的酶解产物经风味蛋白酶作用4 h后,酶解产物的苦味值最低,制备出苦味值为1.28的WG低苦味肽粉。结论 经分步酶解作用后,酶解产物中苦味肽的含量下降;疏水性氨基酸比例的下降和游离氨基酸含量的升高引起苦味肽苦味阈值的增大,共同导致酶解产物苦味值显著降低,该研究为酶解脱苦技术的快速发展和WG活性肽工业化生产提供新的参考。     

凡纳滨对虾虾头协同水解工艺的响应面优化

利用风味蛋白酶协同虾头内源酶水解凡纳滨对虾虾头,制备水解度高、苦味值低的新型虾风味食品基料.以苦味感官为指标,采用响应面法研究pH、温度、底物浓度对产物水解度和苦味的影响.在pH值6.92,温度52.5 ℃,底物浓度为22.25 g/100 mL的条件下,每克虾头添加144 U风味蛋白酶水解3 h,与自溶水解液相比,水解度增加了6.4%,苦味值降低了48.5%,游离氨基酸总量增加了16.8%,疏水性氨基酸总量增加了4.5%.风味蛋白酶可以协同虾头内源酶水解凡纳滨对虾虾头,制备出水解度高、苦味值低的水解液.     

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。     

组合酶对复合骨素酶解液呈味物质的影响

为探究组合酶对牛骨素和鸡骨素的复合骨素酶解液呈味物质的影响,选取四种组合酶(木瓜蛋白酶+风味蛋白酶、菠萝蛋白酶+风味蛋白酶、碱性蛋白酶+风味蛋白酶、复合蛋白酶+风味蛋白酶)制备复合骨素酶解液,测定四种复合骨素酶解液的水解度、游离氨基酸、呈味核苷酸、味精当量(Equivalent umami concentration,EUC)、肽分子量分布等呈味物质指标,并进行对比分析。结果表明:碱性蛋白酶+风味蛋白酶(Alkaline proteinase+Flavourzyme,A+F)和复合蛋白酶+风味蛋白酶(Protamex+Flavourzyme,P+F)酶解液的水解度最大,分别为10.67%和11.27%;对呈味游离氨基酸组成分析发现,A+F酶解液鲜味氨基酸、苦味氨基酸、无味氨基酸含量最高,A+F和P+F酶解液总游离氨基酸含量最高;四种酶解液中肌苷酸较另两种核苷酸含量高,A+F酶解液总核苷酸含量最高;比较四种酶解液味精当量,A+F酶解液EUC值最大;A+F和P+F酶解液中分子量<1000 Da肽段含量最高,制备复合骨素酶解液的呈味效果更好;主成分分析表明A+F组合酶综合得分最高,A+F组合酶为美拉德反应提供丰富反应底物。     

不同蛋白酶制剂对牛肉的酶解效果研究

以复合蛋白酶、木瓜蛋白酶、胰蛋白酶和风味蛋白酶对牛肉进行预处理,测定酶解液的理化指标、多肽分子量分布、游离氨基酸等,筛选出适宜蛋白酶。结果表明,复合蛋白酶在可溶性氮、氨基酸态氮,水解度等指标上优于其他种类蛋白酶制剂。添加木瓜蛋白酶、复合蛋白酶的酶解液在2 KDa～5 KDa、小于1 KDa的多肽分布较高,这两部分多肽对食品风味具有重要作用。添加复合蛋白酶酶解液的氨基酸总量最多,其中鲜味氨基酸含量明显高于其他组别,苦味氨基酸含量低于其他组别。结合酶解液的感官评定结果,复合蛋白酶是比较适宜酶解牛肉的酶制剂。     

响应面法优化紫贻贝蛋白酶解工艺条件

分别采用中性蛋白酶、碱性蛋白酶、酸性蛋白酶、复合蛋白酶、木瓜蛋白酶、胰蛋白酶和风味蛋白酶7种蛋白酶对紫贻贝蛋白的酶解工艺条件进行研究。根据水解度和感官评定的结果,确定中性蛋白酶、复合蛋白酶、木瓜蛋白酶和风味蛋白酶可以作为紫贻贝蛋白酶解的外加蛋白酶。将上述4种蛋白酶进行两两复配,通过试验确定复合蛋白酶与中性蛋白酶按1:1进行复配,可作为紫贻贝蛋白酶解的最适复配酶。采用响应面优化分析得出复配蛋白酶最佳酶解条件为酶解时间2h、pH7、酶解温度50℃、酶添加量0.4%,在此条件进行实验,测得水解度为70.25%,游离氨基酸总量增加了388.46%。     

酶法制备不同水解度高温豆粕水解产物的理化特性研究

采用Alcalase酶和木瓜蛋白酶分别对高温大豆粕进行酶解,通过控制酶解反应得到水解度为5%、10%和15%的6种水解产物,研究两种酶对不同水解度的水解产物理化特性的影响。结果表明,Alcalase酶和木瓜蛋白酶均可产生6种不同分子量范围的水解产物,但各部分比例具有显著差异(P0.5),其平均分子量随水解度的增加逐渐减少,Alcalase酶的水解产物中小于2562 Da小分子量肽所占比例更高。豆粕蛋白的疏水基团在酶解反应中发生暴露与断裂的数量差,导致其表面疏水性随水解度增加呈现先下降再上升的变化,即水解度为10%的表面疏水性最低。zeta电势的绝对值随水解度不断上升,分子间的斥力增大,相同水解度下两种酶对zeta电势的影响并不显著。此外,在pH值为3、5、7和9时,水解产物的溶解性随着水解度的增加而逐渐增高,乳化活性和乳化稳定性则逐渐降低。     

两种酶水解鲢鱼蛋白产物功能性质研究

采用Alcalase 2.4L和Flavourzyme 500L蛋白酶对鲢鱼肉进行水解,研究了水解产物的水解度与体系的pH值对其功能性质的影响。水解产物的溶解度在pH4时最低,碱性时具有较高的溶解度;浊度随pH值变化趋势与溶解度相反,浊度越大,溶解度越小。随水解度的增加,水解产物的乳化活性指数、乳化稳定性、起泡性和泡沫稳定性减小(p<0.05);在水解度相同情况下,水解产物的功能性质取决于所用酶的种类。结果表明,鲢鱼肉蛋白水解产物的功能性质受其水解度和所用酶种类的影响。     

Evaluation of bitterness in enzymatic hydrolysates of soy protein isolate by taste dilution analysis

ABSTRACT:  Although enzymatic hydrolysates of soy protein isolate (SPI) have physiological functionality, partially hydrolyzed SPI exhibits bitter taste depending on proteases and degree of hydrolysis (DH). To determine proteolysis conditions for SPI, it is important to evaluate bitterness during enzymatic hydrolysis. Taste dilution analysis (TDA) has been developed for the screening technique of taste-active compounds in foods. The objectives of the present study were to evaluate bitterness of enzyme-hydrolyzed SPI by TDA and to compare bitterness of SPI hydrolysates with respect to kinds of proteases and DH. SPI was hydrolyzed at 50 °C and pH 6.8 to 7.1 to obtain various DH with commercial proteases (flavourzyme, alcalase, neutrase, protamex, papain, and bromelain) at E/S ratios of 0.5%, 1%, and 2%. The DH of enzymatic hydrolysates was measured by trinitrobenzenesulfonic acid method. The bitterness of enzymatic hydrolysates was evaluated by TDA, which is based on threshold detection in serially diluted samples. Taste dilution (TD) factor was defined as the dilution at which a taste difference between the diluted sample and 2 blanks could be detected. As DH increased, the bitterness increased for all proteases evaluated. Alcalase showed the highest TD factor at the same DH, followed by neutrase. Flavourzyme showed the lowest TD factor at the entire DH ranges. At the DH of 10%, TD factor of hydrolysate by flavourzyme was 0 whereas those by protamex and alcalase were 4 and 16, respectively. These results suggest that TDA could be applied for the alternative of bitterness evaluation to the hedonic scale sensory evaluation.     

ENZYMATIC HYDROLYSIS OF WHEY PROTEINS BY TWO DIFFERENT PROTEASES AND THEIR EFFECT ON THE FUNCTIONAL PROPERTIES OF RESULTING PROTEIN HYDROLYSATES

Whey protein concentrate (WPC 80) was hydrolyzed by Alcalase 2.4 L and Protamex to 5, 10, 15 and 20% degree of hydrolysis (DH). WPC 80 and its hydrolysates were analyzed, compared and used for measuring some functional properties. All hydrolysates were different from WPC 80 in protein, moisture and ash content. Free amino groups and protein solubility increased with increasing DH. The peptides produced by hydrolysis had smaller molecular sizes, and their average molecular weight decreased as the DH increased. Except hydrolysates generated by Alcalase 2.4 L at 5% DH, all others showed poor emulsifying and foaming properties compared with unhydrolyzed WPC 80. Gelation properties of WPC 80 and its hydrolysates were different. The global amino acid compositions did not differ significantly between the different hydrolysates, and they were very close among WPC 80 and its hydrolysates except for Methionine, Glycine, Histidine and Valine.     

Angiotensin-I-converting enzyme inhibitory activity and bitterness of enzymatically-produced hydrolysates of shrimp (Pandalopsis dispar) processing byproducts investigated by Taguchi design

The importance of water-to-substrate ratio, protease type, percent enzyme and incubation time on hydrolysates produced from shrimp processing byproducts was investigated using Taguchi’s L₁₆ (4⁵) experimental design. Protease type significantly (p < 0.05) influenced soluble yield, degree of hydrolysis (DH), angiotensin-I-converting enzyme (ACE) inhibitory activity and bitterness of hydrolysates, while percent enzyme only affected the DH. Hydrolysates produced by Alcalase and Protamex possessed strong ACE inhibitory activity (IC₅₀ = 100–200 μg/ml and 70 μg/ml, respectively), accompanied by high yield, high DH and strong bitterness. Furthermore, ACE inhibition was positively correlated (r² = 0.87) with bitterness of the hydrolysates. Fractionation by size-exclusion chromatography revealed that the bitter substances, which also showed strong ACE inhibition, were <3 kDa in size and contained many hydrophobic residues, including Tyr, Phe, Leu, Ile, Val and Lys. Despite the bitterness, these hydrolysates may have potential health benefits, arising from their potent ACE inhibitory activity.     

Bitterness intensity of soybean protein hydrolysates—chemical and organoleptic characterization

Soluble and isoelectric soluble soybean protein hydrolysates were prepared by Alcalase treatment to a degree of hydrolysis of 3–15%. The bitterness intensity of the hydrolysates obtained was assessed on a five-point scale. The average relative molecular masses of peptides in the soluble hydrolysates (2250-1400) and isoelectric soluble hydrolysates (1313-800) and their hydrophobic peptide fractions (575-400) were determined by the trinitrobenzenesulphonic acid method. The molecular mass distribution of peptides in soluble and isoelectric soluble soybean hydrolysates and their hydrophobic peptide fractions was determined by gel permeation HPLC using a Zorbax Bio Series GF-250 column. The results suggest that the main reason for the bitterness of soybean protein hydrolysates prepared by Alcalase treatment are hydrophobic bitter peptides of relative molecular mass less than 1000.     

Modification of the nitrogen solubility properties of soy protein isolate following proteolysis and transglutaminase cross-linking

The effect of (a) limited hydrolysis [0.5–2.0% degree of hydrolysis (DH)] with Alcalase™, (b) cross-linking with transglutaminase (TGase) and (c) combinations of these modifications on the nitrogen solubility (pH 3–8) of soy protein isolate (SPI) was investigated. Between pH 3.0 and 5.0, SPI hydrolysates, hydrolysates of cross-linked SPI and the cross-linked products of SPI hydrolysates displayed significant (P<0.05) increases in solubility compared to unmodified SPI. Cross-linking pre- or post hydrolysis did not alter the overall trend of increased (P<0.05) solubility relative to the unmodified control at low pH. At 2% DH, cross-linking pre- or post-hydrolysis resulted in greater solubility (P<0.05) than that observed in hydrolysates per se at low pH. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS–PAGE) indicated that the 22 kDa 11S basic polypeptide was relatively resistant to Alcalase hydrolysis and that the 18 and 22 kDa 11S basic polypeptides were not susceptible to TGase cross-linking. The results demonstrate that a combination of enzymatic treatments and the order in which they are applied may have potential for creating novel food ingredients with improved functional properties, especially those properties that are dependant on high solubility at low pH.     

花生蛋白水解产物ACE抑制活性的研究

采用胰蛋白酶、碱性蛋白酶、中性蛋白酶水解花生蛋白,研究了水解过程中水解度的变化,并对水解产物的ACE抑制活性进行了探讨。得出三种酶对花生蛋白的水解作用:碱性蛋白酶>胰蛋白酶>中性蛋白酶。碱性蛋白酶水解产物ACE抑制活性明显高于胰蛋白酶和中性蛋白酶,水解产物的ACE抑制活性高达89.73%,中性蛋白酶水解产物ACE抑制率仅为27.24%。     

Functional Properties and Bitterness of Sodium Caseinate Hydrolysates Prepared with a Bacillus Proteinase

The functional and physicochemical characteristics, and bitterness were evaluated on sodium caseinate hydrolysates generated with a commercial Bacillus proteinase complex. At low degrees of hydrolysis (0.5 and 1.0% DH) the hydrolysates compared to the unhydrolyzed substrate had increased emulsion activity at pH 2 and 4. However, higher DH resulted in lower emulsion activities. At pH 8 and 10, the low DH hydrolysates displayed increased foam expansion and decreased foam drainage compared to the starting substrate. The Bacillus proteinase hydrolysates at higher DH had similar bitterness values to tryptic hydrolysates at equivalent DH. However, the Bacillus hydrolysate at 0.5% DH was less bitter (p<0.05) than the tryptic hydrolysate at the same DH.