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。     

蛋白质酶解产物苦味的形成及脱除的研究进展

蛋白质酶解物苦味的苦味是长期困扰其应用的问题，本文对苦味肽的产生、化学组成及脱除苦味的方法进行了综述。     

食品蛋白质降血压肽(ACEIP)的开发研究

降血压肽(ACEIP)主要来源于食品蛋白质，其制备过程包括蛋白质的提取、酶水解、降血压肽的分离、脱苦、干燥、检测等环节。介绍和分析了蛋白酶及其酶解条件的选择、活性肽的分离、苦味的脱除及活性的测定等开发降血压肽的关键技术和难点。     

小麦面筋蛋白深度酶解过程中呈味物质变化趋势的研究

以麸皮和面粉制曲,利用米曲霉生长过程中产生的复合酶,对小麦面筋进行深度酶解,研究了产物的呈味变化趋势及酶解过程中蛋白质、氨基酸、总酸、总糖、肽的释放规律。酶解产物的鲜味、酸味随着酶解时间的延长不断增强,苦味、甜味不断下降。蛋白质回收率、氨基酸转化率、总酸在酶解的过程中逐渐增加,但是36h后增速均趋于平缓,总糖的含量不断下降。随着酶解时间的延长,大分子肽渐渐分解成小分子肽,主要表现为1000～5000u分子段的肽的含量减少和小于1000u分子段的肽的含量增加。     

小麦面筋蛋白深度酶解过程中呈昧物质变化趋势的研究

以麸皮和面粉制曲,利用米曲霉生长过程中产生的复合酶,对小麦面筋进行深度酶解,研究了产物的呈味变化趋势及酶解过程中蛋白质、氨基酸、总酸、总糖、肽的释放规律。酶解产物的鲜味、酸味随着酶解时间的延长不断增强,苦味、甜味不断下降。蛋白质回收率、氨基酸转化率、总酸在酶解的过程中逐渐增加,但是36h后增速均趋于平缓,总糖的含量不断下降。随着酶解时间的延长,大分子肽渐渐分解成小分子肽,主要表现为1000-5000U分子段的肽的含量减少和小于1000u分子段的肽的含量增加。     

不同酶水解马氏珍珠贝蛋白的特性研究

本研究采用Alcalase2.4L、Protamex、Papain、PTN6.0S、Flavorzyme500MG、Neutrase和内源酶水解马氏珍珠贝肉蛋白,探讨了各种酶的酶解特性、产物的氨基酸组成及肽相对分子质量分布规律和呈味特征的差异.结果表明,PTN6.0S酶解产物的蛋白质利用率、肽得率和水解度均较高,而内源酶均最低.各酶解产物中,谷氨酸,精氨酸和天冬氨酸的含量均较高.同一氨基酸在不同酶解产物中回收率存在较大差异,色氨酸、苏氨酸、脯氨酸和天冬氨酸在马氏-爹珠贝蛋白酶解产物中主要以多肽的形式存在,各酶解产物中氨基酸的回收率和释放率较高的氨基酸与各酶的主要作用位点有很好的对应关系;各种酶均能不同程度地降解马氏珍珠贝肉蛋白和相对分子质量大于5000Da的肽断.各酶水解产物肽相对分子质量分布差异主要体现在3000以下.在各酶解产物中,Papain酶解产物风味最佳,鲜味最高,苦味和腥味最低.     

扇贝裙边酶解过程中呈味组分的变化规律研究

本研究以扇贝裙边为原料,利用复合蛋白酶对扇贝裙边进行酶解,在确定扇贝裙边最佳酶解温度的基础上,探索了扇贝裙边酶解液在不同酶解时间下的呈味特点,并探讨了不同酶解液中呈味分子的变化规律,探明不同酶解时间下扇贝裙边酶解液的呈味规律。结果表明不同酶解时间制备酶解液的滋味存在显著性差异,其中8 h的扇贝裙边酶解液鲜味强度最高(9.32分),而12 h酶解液的苦味(7.33分)和饱满度(8.33分)最强。主要是因为酶解时间对酶解液鲜味氨基酸和苦味氨基酸的含量及比例存在较大影响,当酶解8 h时,扇贝裙边酶解液中鲜味氨基酸比例最高(46.80%),而苦味氨基酸比例最低(51.67%);此外,肽分子分布结果显示8h酶解液中5000 u的肽段(对呈味贡献小)和180 u的肽段(苦涩味明显)比例较低,可能是8 h扇贝裙边酶解液取较好的鲜味和饱满度,较低苦味的主要原因。本研究通过研究扇贝呈味组分在酶解过程中的变化规律,为工业上利用扇贝裙边制备高品质呈味基料提供理论基础和指导。     

苦味肽的形成及脱苦方法的研究

蛋白质水解后其溶解性、热稳定性以及营养特性得到了改善，但同时却形成了大量的苦味肽。本文综述了苦味肽的形成、影响苦味的主要因素及主要的脱苦方法。     

响应面法优化骨素酶解制备可溶性肽工艺研究

利用响应面法分析了酶解骨素制备可溶性肽的工艺,以酶解温度、时间、pH及酶用量为影响因素,酶解液中可溶性肽含量、酶解液的苦味和鲜味为优化指标。结果显示,酶解温度和pH对可溶性肽含量、酶解液鲜味有显著影响(p0.05);拟合模型验证结果表明,所得模型在预测可溶性肽含量时具有一定可靠性;酶解骨素的最优工艺为温度60℃、时间2h、pH6、酶用量10000U/g,此时可溶性肽含量最高,酶解液的鲜味较强,无苦味。     

超声辅助酶解对龙头鱼蛋白肽理化性质及风味特性的影响

目的:解决龙头鱼酶解蛋白肽鲜味低、苦味重、风味不良等问题。方法:利用超声辅助酶解法制备龙头鱼蛋白肽，以木瓜蛋白酶作为催化剂，研究不同超声功率(0,120,240,480,600 W)条件下龙头鱼蛋白肽粒径、水解度、可溶性多肽含量、相对分子质量分布、游离氨基酸含量以及电子舌风味信号的变化，进而探究不同超声强度对龙头鱼蛋白肽结构及风味特性的影响。结果:超声强度360 W、超声时间1 h,水浴温度55℃以及水浴时间4 h的条件下，蛋白质水解度达到19.29%,可溶性多肽含量为0.57 mg/mL,超声辅助显著降低了龙头鱼蛋白肽的粒径和相对分子质量，相对分子质量<3 000的组分占比从67%增加至82%。此外，龙头鱼蛋白肽中，苦味氨基酸占主要地位，其次是甜味和鲜味氨基酸。超声辅助酶解蛋白肽中鲜味游离氨基酸含量的增幅最大，因此，超声辅助酶解通过增加鲜味氨基酸的含量降低了龙头鱼蛋白肽的苦味。结论:超声辅助可以提高龙头鱼蛋白的酶解作用，进而改善龙头鱼多肽的理化性质及风味特性。     

Characteristic Property of Low Bitterness in Protein Hydrolysates by a Novel Soybean Protease D3

ABSTRACT:  Enzymatic hydrolysis is 1 means of improving the functional properties of food protein; however, in most cases, bitter peptides are generated by such treatment, and the resulting product is therefore not acceptable as a food ingredient. We have already reported a novel cysteine protease, D3, purified from germinating soybean cotyledons. Because of its substrate specificities, most hydrophobic amino acid residues in the hydrolysate are presumed not to be located at the peptide termini. It was therefore expected that protein hydrolysate by protease D3 would taste less bitter than other enzymatic hydrolysates. The objective of this study was to demonstrate the low bitterness of protein hydrolysates by protease D3. For that purpose, soy protein and casein hydrolysates were prepared with treatment of protease D3, subtilisin, pepsin, trypsin, and thermolysin, respectively. The bitterness of these hydrolysates was evaluated by measuring points of subjective equality (PSE). The PSE value demonstrated that the protein hydrolysates by protease D3 were significantly less bitter than the other enzymatic hydrolysates, indicating that the products had a taste mild enough to be acceptable as a less-bitter peptide food ingredient. These results suggested that a prominent feature of protease D3 was its capacity to produce less-bitter peptides. Therefore, it is thought that protease D3 could be applied to produce protein hydrolysates for use as ingredients in a variety of food products.     

Valorisation of protein hydrolysates from animal by-products: perspectives on bitter taste and debittering methods: a review

Conversion of animal by-products to high value-added food ingredients is one of the top trends in the slaughter industry. Enzymatic hydrolysis of animal by-products can generate protein hydrolysates, which provides an opportunity for effective utilisation. However, bitterness of protein hydrolysate is a major undesirable aspect for various applications. In this review, the current knowledge on protein hydrolysates from animal by-products is briefly reviewed. The structural features of bitter peptides and bitter taste receptors are summarised. Moreover, the potential approaches for debittering protein hydrolysates are highlighted, including exopeptidase treatment, Maillard reaction, plastein reaction and encapsulation. In addition, the current debittering strategies and challenges are also discussed. This article presents some opportunities to utilise protein hydrolysates from animal by-products and their debittering methods.     

实用类蛋白反应的作用机制及其对海洋源蛋白修饰的研究进展

类蛋白反应作为一种可以修饰生物活性肽的新方法,已经成为蛋白质食品研究的热点。海洋生物含有丰富的生物活性成分,其含有的蛋白肽具有多种人体代谢和生理调节功能。然而蛋白肽存在生物利用度有限、酶解液味苦等问题。本文在阐述类蛋白反应过程和机制的基础上,对类蛋白反应修饰提高海洋源活性蛋白肽的生物活性、改善蛋白的加工特性及减少蛋白酶解液苦味等研究进展进行了详细的介绍,以期为海洋源蛋白的深度利用以及高值化研究提供理论参考。     

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 Production,Bioactivity, and Bitterness of Chickpea (Cicer arietinum) Peptides

Chickpeas are inexpensive, protein rich (approximately 20% dry mass) pulses available worldwide whose consumption has been correlated with positive health outcomes. Dietary peptides are important molecules derived from dietary proteins, but a comprehensive analysis of the peptides that can be produced from chickpea proteins is missing in the literature. This review provides information from the past 20 years on the enzymatic production of peptides from chickpea proteins, the reported bioactivities of chickpea protein hydrolysates and peptides, and the potential bitterness of chickpea peptides in food products. Chickpea peptides have been enzymatically produced with pepsin, trypsin, chymotrypsin, alcalase, flavorzyme, and papain either alone or in combination, but the sequences of many of the peptides in chickpea protein hydrolysates remain unknown. In addition, a theoretical hydrolysis of chickpea legumin by stem bromelain and ficin was performed by the authors to highlight the potential use of these enzymes to produce bioactive chickpea peptides. Antioxidant activity, hypocholesterolemic, and angiotensin 1‐converting enzyme inhibition are the most studied bioactivities of chickpea protein hydrolysates and peptides, but anticarcinogenic, antimicrobial, and anti‐inflammatory effects have also been reported for chickpea protein hydrolysates and peptides. Chickpea bioactive peptides are not currently commercialized, but their bitterness could be a major impediment to their incorporation in food products. Use of flavorzyme in the production of chickpea protein hydrolysates has been proposed to decrease their bitterness. Future research should focus on the optimization of chickpea bioactive peptide enzymatic production, studying the bioactivity of chickpea peptides in humans, and systematically analyzing chickpea peptide bitterness.     

Exopeptidases and their application to reduce bitterness in food: a review

When exopeptidases catalyze hydrolysis of peptide bonds, the product(s) may have a less bitter taste, and the free amino acids or small peptides formed may function in food as pleasant-tasting flavor compounds or as flavor precursors. There are several classes of exopeptidase based on specificity for hydrolysis of synthetic substrates. Exopeptidases in food-stuff may be of natural origin or may be extrinsic, that is, produced by microorganisms or parasites. Exopeptidases used to modify foods are also becoming increasingly available in the industrial enzyme market. Exopeptidases contribute to a variety of quality changes in postharvest fruit, meats, and food fermentations. Foodstuff impacted by these enzymes during processing include cocoa, beer, aged and cured meat products, koji, fish sauce, ripened cheeses, and protein hydrolysates. An important role of exopeptidases in food is the hydrolysis of hydrophobic, bitter peptides. The relationship between peptide structure and sensory transduction/receptor models is discussed. Research on the use of exopeptidases to reduce bitterness is reviewed.     

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。     

Enzymatic hydrolysis of soy proteins and the hydrolysates utilisation

Soy proteins are very important protein source for human being and livestock. Enzymatic hydrolysis of soy protein can enhance or reduce its functional properties and improve its nutritious value. Soy protein hydrolysates were primarily used as functional food ingredients, flavour and nutritious enhancers, protein substitute, and clinical products. Conditions for hydrolysis were usually mild, whereas recently high pressure treatment attracted more interest. Degree of hydrolysis (DH) was usually between 1% and 39.5%. The main problem associated with proteolytic hydrolysis of soy protein was production of bitter taste, hydrolysates coagulation and high cost of enzymes. Bitterness reduction can be achieved by control of DH, selective separation of bitter peptides from hydrolysates, treatment of hydrolysates with exo‐peptidases, addition of various components [adenosine monophosphate (AMP), some amino acids, monosodium glutamate (MSG), etc.] to block or mask the bitter taste, and modification of taste signalling. Hydrolysates coagulation can be resolved by selecting appropriate enzymes and by applying immobilisation technology the production cost can be reduced. Enzymatic hydrolysis also enhances bioactivity of soy proteins through conversion of glycosides to aglycones, increasing antioxidant and immunoregulatory properties. Finally, future works have been discussed.