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1.
乳扇、乳饼和奶渣是云南民族传统乳制品,具有历史悠久、特色鲜明、蛋白质含量高等特点。本研究基于模拟胃肠道消化,采用液相色谱-串联质谱、氨基酸分析仪研究乳扇、乳饼和奶渣的肽谱及游离氨基酸含量,并借助蛋白肽数据库分析潜在的生物活性肽。结果表明,乳扇蛋白肽221 条,分子质量集中在500~750 Da,潜在的功能活性肽包括血管紧张素转化酶抑制肽5 条、免疫调节肽4 条、抗氧化肽3 条,抗血栓肽2 条和抗菌肽1 条。乳饼蛋白肽225 条,分子质量集中在750~1 000 Da,功能活性肽包括抗氧化肽2 条。奶渣蛋白肽222 条,分子质量集中在750~1 000 Da,功能活性肽包括免疫调节肽2 条、抗氧化肽2 条和抗血栓肽1 条。3 种乳制品中的蛋白肽主要源于β-和α-酪蛋白。乳扇、乳饼和奶渣经模拟消化液消化后其游离氨基酸含量分别为9.45、10.7 g/100 g和9 g/100 g,其中,中性氨基酸含量最高,其次依次为碱性氨基酸和酸性氨基酸。本研究揭示了乳扇、乳饼和奶渣的生物学价值,可为功能活性产品的研发提供科学依据。  相似文献   

2.
以植物乳杆菌(Lactobacillus plantarum)CD101和模仿葡萄球菌(Staphylococcus simulans)NJ201作为混合发酵剂制作发酵香肠,以自然发酵为对照。通过测定理化指标、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecylsulfate-polyacrylamidegelelectrophoresis,SDS-PAGE)、多肽含量、游离氨基酸含量等指标研究混合发酵剂对发酵香肠蛋白质降解情况,并以体外1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除活性、2,2’-联氮双(3-乙基苯并噻唑啉-6-磺酸)(2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid),ABTS)阳离子自由基清除活性及铁离子还原/抗氧化能力(ferric ion reducing antioxidant power,FRAP)值评价发酵香肠粗肽及小分子多肽(3 kDa)的抗氧化能力。结果表明:混合发酵剂接种组非蛋白氮含量显著高于对照组(P0.05);根据SDS-PAGE结果分析,2组发酵香肠的肌浆蛋白和肌原纤维蛋白均发生降解,接种组的蛋白降解程度高于对照组,尤其是在低分子质量(20 kDa)条带处出现明显累积;接种组粗肽及小分子多肽的DPPH自由基清除活性、ABTS阳离子自由基清除活性及FRAP值均显著高于对照组(P0.05),其中小分子多肽可能是发酵香肠多肽抗氧化能力的主要贡献者;同时发酵剂促进发酵香肠中游离氨基酸的释放。接种该混合发酵剂制作发酵香肠能促进蛋白质降解,得到更多具有抗氧化活性的多肽,从而有助于通过内源性抗氧化肽抑制发酵香肠的氧化,降低生产成本,延长货架期。  相似文献   

3.
大河乌猪火腿在发酵和后熟过程中蛋白质降解可产生丰富的生物活性肽。为探究大河乌猪火腿中是否存在α-葡萄糖苷酶抑制肽及其活性,以大河乌猪火腿为研究对象,通过超滤分离方法制备了不同分子质量的火腿肽;以α-葡萄糖苷酶抑制率为指标,采用肽组学结合生物信息学分析方法对火腿肽进行鉴定、筛选和活性研究。结果表明:分子质量小于3 kDa的大河乌猪火腿肽具有良好的α-葡萄糖苷酶抑制活性。从大河乌猪火腿中共鉴定出143条主要来源于肌球蛋白、肌钙蛋白和β-烯醇化酶的肽序列,进一步筛选出的肽段IEEALGDK具有良好的α-葡萄糖苷酶抑制活性(IC50值为1.42 mg/mL)。BIOPEP-UWM数据库检索结果显示,肽段IEEALGDK为新型的生物活性肽。肽的稳定性研究表明,肽段IEEALGDK具有较好的热稳定性、耐酸碱性和胃肠道消化稳定性。分子对接结果显示,肽段IEEALGDK主要通过氢键和疏水相互作用占据α-葡萄糖苷酶的活性残基位点Arg594、Arg727、Arg799和Arg467发挥活性作用。大河乌猪火腿的肽段IEEALGDK为新型生物活性肽,具有良好的α-葡萄糖苷酶抑制活性,研...  相似文献   

4.
利用液相色谱-串联质谱(liquid chromatography-tandem mass spectrometry,LC-MS/MS)的方法,对4种酸乳(优味舒风味发酵乳、风味发酵乳、初乳24+K风味发酵乳和十天风味发酵乳)的肽谱进行分析,分别鉴定到152、125、364条和274条肽段。利用血管紧张素转换酶(angiotensin converting enzyme,ACE)的抑制活性比较以上4种酸乳的生物活性,在相同质量浓度2.0 mg/L时,4种酸乳ACE抑制率分别为46.84%、42.75%、77.41%和63.84%。初乳24+K风味发酵乳含有肽的数量大、ACE抑制活性高可能与其独特的原料成分相关。对4种酸乳的肽谱及ACE抑制活性的考察,为进一步研究酸乳肽的功能特点和ACE抑制活性提供了理论依据。  相似文献   

5.
研究红曲霉对奶酪成熟过程中生物活性肽,尤其血管紧张素转换酶(angiotensin-converting enzyme,ACE)抑制肽的影响。以不接种红曲霉的奶酪为对照组,研究奶酪成熟过程中的可溶性氮含量、α-葡萄糖苷酶抑制活性和ACE抑制活性,对红曲霉奶酪肽谱进行鉴定,利用生物信息学和分子对接等方法对潜在的活性肽进行筛选,并对筛选肽进行合成和评价。结果表明,相比于对照组,在pH 4.6和三氯乙酸条件下,奶酪成熟结束时的可溶性氮相对含量分别提高57.98%和38.34%,α-葡萄糖苷酶抑制活性和ACE抑制活性分别提高32.21%和73.98%。在红曲霉奶酪提取物中共鉴定到1 796种肽,其中51条ACE抑制肽、28条抗氧化肽、7条降血糖肽等。筛选到两种ACE抑制肽YPFPGPI和FPEVFGK,半抑制浓度分别为207.31μmol/L和410.61μmol/L,酶抑制类型均是非竞争性抑制。该研究提供了一种快速筛选ACE抑制肽的方法,证明奶酪中添加红曲霉能够有效提高ACE抑制活性,为挖掘红曲霉奶酪的功能特性潜力提供了参考。  相似文献   

6.
甲鱼蛋白酶解产物体外ACE抑制和抗氧化活性研究   总被引:1,自引:0,他引:1  
目的研究甲鱼蛋白酶解产物的血管紧张素转换酶(ACE)的抑制活性和抗氧化性.方法以体外活性(ACE抑制率、·0H、O2-·和DPPH·清除率)为指标,通过正交试验确定木瓜蛋白酶的最佳酶解条件;用Sephadex G-25分离、提纯.检测各组分的ACE抑制活性和抗氧化活性.结果木瓜蛋白酶酶解甲鱼蛋白制备ACE抑制肽的最佳酶解条件为温度60℃、pH 7.0、酶解时间3 h、加酶量1.0%;在此酶解条件下酶解产物的ACE抑制率为99.96%,获得了ACE抑制活性较高的组分F4和F1,其ACE抑制率分别为67.27%、52.73%;制备抗氧化肽的最佳酶解条件为温度60℃、pH 5.0、时间4h、加酶量0.6%,得到抗氧化活性最高的组分F2,该组分的·OH、O2-·和DPPH·清除率分别为10.39%、33.00%、17.68%,其相对分子质量范围集中在307左右.结论甲鱼多肽具有较强的体外ACE抑制活性和抗氧化活性.  相似文献   

7.
为优化酶解法制备绵羊乳酪蛋白ACE抑制肽的工艺条件以及筛选和鉴定一种新的ACE抑制肽,选用5种蛋白酶水解酪蛋白,以水解度、分子质量分布和ACE抑制率为指标筛选最适蛋白酶,采用单因素和响应面试验优化工艺,采用三合一质谱仪(Orbitrap Fusion Lumos Tribrid MS)方法鉴定分子质量小于3 ku组分的氨基酸序列,筛选潜在ACE抑制肽,进行人工合成,测定其IC50值。采用Linewaver-Burk作图确定酶抑制动力学,结合分子对接解析肽段的抑制机制。结果表明:碱性蛋白酶水解酪蛋白的最佳条件为pH 6、底物含量8%、酶添加量4%、温度55 ℃、水解时间90 min,此时酪蛋白水解液ACE抑制率为99.1%。验证具有ACE抑制活性的肽段10条,筛选出一条新颖的降血压肽——LFRQFY(源自αs1-酪蛋白),其ACE抑制活性的IC50为(7.9±1.7)μmol/L,酶抑制动力学为混合抑制模式。分子对接结果表明:LFRQFY能与ACE的氨基酸残基Ala354(活性口袋S1)、His353(活性口袋S2)形成氢键,具有显著的体外降血压活性。  相似文献   

8.
通过使用从传统酸马乳中分离纯化和筛选出的乳酸菌与酵母菌共发酵马乳,获得具有血管紧张素转换酶(angiotensin-converting enzyme,ACE)抑制活性的生物活性肽。经过发酵性能和生长性能测试后筛选到发酵马乳ACE抑制率为38.67%的乳酸乳球菌(Lactococcus lactis)L8和ACE抑制率为51.33%解脂耶氏酵母(Yarrowia lipolytica)Y7进行组合发酵,单因素试验和响应面试验优化发酵条件后发酵马乳的ACE抑制率达到80.67%。与单菌发酵相比,ACE抑制率显著提高。对共发酵产物进行分离纯化,得到了ACE抑制率为86%的活性组分,发酵马乳是分离制备ACE抑制肽的潜在来源。  相似文献   

9.
豆豉功能性的研究   总被引:13,自引:3,他引:10  
本文对不同品种豆豉的抗氧化、α-葡萄糖苷酶抑制和血管紧张素转换酶(ACE)抑制效果进行了比较,并对埃及曲霉纯种发酵曲中ACE抑制剂与ACE和肠胃蛋白酶预混合保温后的抑制活性进行分析,结果表明豆豉曲提取液具有较好的抗氧化和α-葡萄糖苷酶抑制效果;与ACE预混合保温后ACE抑制活性变化不大,而被不同的肠胃蛋白酶消化后抑制活性均有较大的提高,为前药型抑制剂。  相似文献   

10.
利用鲍鱼加工副产物外套膜制备具有抑制血管紧张素转移酶(angiotensin converting enzyme,ACE)活性的生物活性肽,为鲍鱼加工副产物的高值化利用提供新思路。采用从凡纳滨对虾消化腺中制备的丝氨酸蛋白酶,酶解皱纹盘鲍外套膜蛋白,以酶解物ACE抑制活性为评价指标优化条件。酶解液通过3 kDa超滤膜后,活性组分经过Superdex peptide 10/300 GL凝胶过滤柱和反相高效液相色谱(reversed-phase high performance liquid chromatography,RP-HPLC)分离纯化,利用液相色谱-质谱(liquid chromatography tandem mass spectrometry,LC-MS/MS)鉴定纯化肽的氨基酸序列。结果显示,酶解液的ACE抑制率为13.42%,经3 kDa超滤膜初步分级后,相同浓度下,小于3 kDa的组分ACE抑制率为53.25%。经凝胶过滤柱和反相高效液相色谱柱进一步分离后,从ACE抑制活性最高的峰中鉴定得到5个肽段,序列为LGDSFYYGK、LVNEVTEFAK、VDEVGGEALGR、MFLSFPTTK、VATVSLPR,说明活性峰是多肽混合物。本研究利用鲍鱼外套膜制备ACE抑制肽,可以为鲍鱼及对虾加工副产物的高值化利用提供理论参考。  相似文献   

11.
The aim of the present study was to elucidate previous findings showing that peptide fractions isolated from yoghurt had antioxidant effects. Therefore, peptides and free amino acids released during fermentation of milk were characterised. Yoghurt samples were stripped from sugars and lactic acid and subsequently fractionated by ultra filtration using membranes with cut off sizes of 30, 10 and 3 kDa. The peptides in these fractions were identified by LC–MS/MS. The identified peptides comprised a few N-terminal fragments of αs1-, αs2-, and κ-casein, and several fragments from β-casein. Almost all the peptides identified contained at least one proline residue. Some of the identified peptides included the hydrophobic amino acid residues Val or Leu at the N-terminus and Pro, His or Tyr in the amino acid sequence, which is characteristic of antioxidant peptides. In addition, the yoghurt contained a considerable amount of free amino acids such as His, Tyr, Thr and Lys, which have been reported to have antioxidant properties. Thus, our findings confirm that the antioxidant effects of the peptide fractions from yoghurt are due to the presence of certain peptides and free amino acids with recognised antioxidant activity in these fractions.  相似文献   

12.
通过检测乳酸菌对产品感官、微生物、pH值、水分活度、水分含量、生物胺含量、蛋白质降解指数、游离氨基酸含量指标的影响,研究乳酸菌发酵剂对羊肉风干香肠品质的影响。结果表明,添加乳酸菌发酵剂对产品的感官性质具有显著的影响,可以改善产品的色泽和风味,对产品的微生物数量、pH值、生物胺含量、蛋白质降解指数和氨基酸含量都有显著的影响,而对产品的水分含量和水分活度影响不大,通过乳酸菌的发酵作用,乳酸菌发酵剂实验组最终产品pH值为5.18,蛋白质降解指数为17.97%,氨基酸总量达到3 933.67 mg/100 g,都要高于空白组产品。因此,通过添加乳酸菌,可以显著改善产品的感官品质和营养品质,同时降低安全隐患。  相似文献   

13.
丁苗  李成龙  刘淑贞  周才琼 《食品科学》2015,36(19):204-210
对猪肉发酵过程中蛋白质及其降解产物进行分析。结果表明:猪肉在发酵过程中蛋白氮含量随发酵时间延长呈下降趋势,非蛋白氮和氨态氮含量随发酵时间的延长而增加,多肽氮含量呈先升高后降低趋势,在发酵20 d时达最大值(0.227%);发酵20 d酸肉多肽具有最强的体外血管紧张素转化酶(angiotensin I-converting enzyme,ACE)抑制活性,ACE抑制率达74.35%,IC50为2.75 mg/mL;采用超滤、D101型大孔树脂、葡聚糖凝胶对发酵20 d的酸肉多肽进行分离纯化,分离得到的F3组分有较强的ACE抑制活性,IC50为0.90 mg/mL,肽含量为86.54%,氨基酸组成分析显示水解后增加最多的氨基酸是脯氨酸(7.08 倍)和酪氨酸(3.26 倍),谷氨酸、组氨酸、天冬氨酸、苯丙氨酸和丙氨酸占全部肽中氨基酸总量的49.09%,构成肽的疏水性氨基酸、芳香族氨基酸和支链氨基酸分别占39.35%、10.69%和13.65%;反相高效液相色谱显示F3组分主要由9 个峰组成,有待进一步的纯化。  相似文献   

14.
Water-soluble extracts from Gouda-type cheese in a 0.05M sodium citrate buffer at pH 4.0 were fractionated by high-pressure liquid chromatography (HPLC) before and after ripening for 1, 2, and 3 months. Three major peptides were isolated from each sample of extract, but size of the peaks increased with ripening period. The amino acid compositions of these peptides were similar to fragments of αsl-casein, i.e., αsl-CN(fl-9), αsl-CN(fl-13) and αsl-CN(fl-14). αsl-CN(fl-23) was hydrolyzed by cellular proteases of Streptococcus cremoris H61; seven main peptides including αsl-CN(fl-9), αsl-CN(fl-13) and αsl-CN(fl-14) were isolated and characterized by HPLC. This suggests that hydrolysis of αsl-CN(fl-23) by lactic acid bacterial proteinase is one of the main pathways of αsl-casein degradation during Gouda-type cheese ripening.  相似文献   

15.
In the current study, three different types of cheese, cheddar, feta, and Roquefort, were screened to determine the variations in phenolic-linked antioxidant activity and the potential to inhibit key enzymes relevant to type 2 diabetes and related hypertension. The cheese samples were assayed for total phenolic content, related antioxidant activity, and inhibition of α-glucosidase, pancreatic α-amylase inhibitory activity, and the angiotensin-converting enzyme (ACE)-I inhibitory activity. The three fungal-enriched Roquefort cheese samples had the highest total phenolic content. The phenolic content in the herb cheese was slightly but not significantly higher compared to plain cheese. Roquefort cheese samples had the highest antioxidant-linked DPPH (free radical) scavenging activity and as expected DPPH radical scavenging activity was higher in the herb cheese compared to plain cheese. All samples had some α-glucosidase and α-amylase inhibitory activities, with cranberry-enriched cheese having the highest activities. However, no correlation to soluble phenolic content was observed. All the cheese samples had very high anti-ACE-I inhibitory activity, indicating no correlation to phenolic content and activity was even high in 10× diluted samples. The highest ACE-I inhibitory activity was observed in plain and herb-enriched cheddar cheese as well as cranberry-enriched cheese. These studies indicate that cranberry-enriched cheese had the best potential for inhibition of α-glucosidase and α-amylase relevant for type 2 diabetes management, whereas any cheese product had potential for ACE-I inhibition linked to hypertension management, indicating likely the role of other factors such as peptides from cheese fermentation.Industrial relevanceThis research is focused on screening of different types of commercial plain, herbal, fruit, and fungal-enriched to provide a strong biochemical rationale for further design of functional cheese products for anti-type 2 diabetic and relevant hypertension management. A better understanding of these functional attributes provides a strong biochemical rationale for design in vivo and clinical studies from which right design of functional food can be established.  相似文献   

16.
为研究榛子粕抗氧化肽的抗氧化性和氨基酸组成的关系,本实验通过中性蛋白酶水解榛子粕蛋白得到榛子粕抗氧化肽,以DPPH自由基清除率为指标,采用超滤分离和葡聚糖凝胶纯化,获得DPPH自由基清除率高的榛子粕抗氧化肽P2,采用超高压液相色谱串联四级杆质谱(LC-MS/MS)测定榛子粕抗氧化肽的氨基酸序列。P2中7个短肽的氨基酸序列分别为His-ILe/Leu-Pro(362 Da)、ILe/Leu-Val-Asp-Glu(475 Da)、Thr-Pro-Pro-His-Lys(588 Da)、His-ILe/Leu-His-Ser-Ala-Thr(662 Da)、Cys-His-Ser-ILe/Leu-Met-ILe/Leu(701 Da)、Thr-His-Ala(327 Da)、Phe-ILe/Leu(279 Da)。由此得出结论,疏水性氨基酸和芳香性氨基酸可以提高榛子粕抗氧化肽的抗氧化性。  相似文献   

17.
接种总状毛霉(Mucor racemosus)对鱼豆腐进行前发酵研制鱼腐乳,对前发酵过程中氨基酸态氮的变化进行研究,利用十二 烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)观察蛋白质的降解情况。并以蛋白酶活力为评价指标,采用响应面试验优化鱼腐乳前 发酵工艺。 结果表明,发酵至第4天氨基酸态氮含量上升至0.23%,前发酵过程中大部分蛋白质被分解为小分子肽类。 最佳前期发酵 工艺条件为发酵时间103 h,发酵温度27 ℃,菌悬液接种量5.3%,此优化条件下,蛋白酶活力可达51.48 U/g。  相似文献   

18.
为探究产蛋白酶乳酸菌在风干牛肉发酵过程中对肌原纤维蛋白的影响。以牛肉为研究对象,分别接种乳酸乳球菌(S-1)、格氏乳球菌(S-2)、戊糖片球菌(S-3)制备风干牛肉,以不添加发酵剂的样品为对照组(CK),采用理化分析和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(sodium dodecyl sulfate polyacrylamide gel electrophoresis,SDS-PAGE)技术检测风干牛肉发酵过程中肌原纤维蛋白降解情况。结果表明,在发酵和贮藏阶段,非蛋白氮、氨基酸态氮、TCA-可溶性肽含量随着发酵时间延长而逐渐升高,且接种组显著高于对照组(P<0.05),其中S-1组和S-2组样品中非蛋白氮、氨基酸态氮、TCA-可溶性肽含量均显著高于S-3组(P<0.05)。各组样品中氨基酸态氮含量均有增加,其中S-1组氨基酸态氮增加最多,达到577.68 mg/100 g,但实验组间无显著差异(P>0.05)。S-1组和S-2组样品中总游离氨基酸的含量显著增加(P<0.05),SDS-PAGE结果显示接种组中蛋白质降解程度明显高于对照组。在风干牛肉的制作过程中,接种产蛋白酶乳酸菌可以明显提高蛋白质的降解程度,三株乳酸菌中S-1组对蛋白质的降解作用最明显。  相似文献   

19.
Effect of pH and calcium concentration on proteolysis in mozzarella cheese   总被引:1,自引:0,他引:1  
Low-moisture Mozzarella cheeses (LMMC), varying in calcium content and pH, were made using a starter culture (control; CL) or direct acidification (DA) with lactic acid or lactic acid and glucono-delta-lactone. The pH and calcium concentration significantly affected the type and extent of proteolysis in Mozzarella cheese during the 70-d storage period at 4 degrees C. For cheeses with a similar pH, reducing the calcium-to-casein ratio from -29 to 22 mg/g of protein resulted in marked increases in moisture content and in primary and secondary proteolysis, as indicated by polyacrylamide gel electrophoresis and higher levels of pH 4.6- and 5%-PTA-soluble N. Increasing the pH of DA cheeses of similar moisture content, from approximately 5.5 to 5.9, while maintaining the calcium-to-casein ratio almost constant at approximately 29 mg/g, resulted in a decrease in primary proteolysis but had no effect on secondary proteolysis. Comparison of CL and DA cheeses with a similar composition showed that the CL cheese had higher levels of alpha(s1)-CN degradation, pH 4.6- and 5%-PTA-soluble N. Analysis of pH 4.6-soluble N extracts by reverse-phase HPLC showed that the CL cheese had higher concentrations of compounds with low retention times, suggesting higher concentrations of low molecular mass peptides and free amino acids.  相似文献   

20.
以大豆粕为原材料,利用超声辅助酶解技术、超滤-?KTA层析相结合的方法分离纯化获取豆粕酶解产物中血管紧张素转化酶(angiotensin-converting enzyme,ACE)抑制肽,对其分子质量分布进行研究,后通过质谱分析与分子对接技术鉴定并筛选出ACE抑制活性肽的氨基酸序列,经固相合成肽序列,检测其ACE抑制肽的活性并基于分子对接技术探索其抑制机制。结果表明:经超声辅助酶解提取获得的豆粕肽分子质量主要分布在6 000 Da以下;根据分子对接的最低预测自由能筛选出的GVRP(-8.44 kcal/mol)和IIVTP(-9.04 kcal/mol)可以抑制ACE活性,半抑制浓度(50% inhibitory concentration,IC50)分别为(84±0.06)、(77±0.08)μmol/L;分子对接结果表明:GVRP、IIVTP能够与ACE的活性口袋S1、S1′、S2形成氢键相互作用,共有的过近接触(3.5 ?范围内)ACE氨基酸残基为His513、Ala354和Glu384。本研究基于串联质谱与分子对接技术,建立从混合多肽中快速鉴定、筛选活性多肽的方法,探究活性多肽与ACE稳定结合并体现其ACE活性的抑制机制,为后续的深入研究提供一定参考。  相似文献   

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