Novel insight into the formation and inhibition mechanism of dipeptidyl peptidase-Ⅳ inhibitory peptides from fermented mandarin fish(Chouguiyu)

Fermented foods are a potential source to produce novel dipeptidyl peptidase-IV inhibitory peptides(D4IPs).In this study,the fermented mandarin fish(Chouguiyu)was used to screen D4IPs and their formation mechanism was studied by metagenomics and peptidomics.A total of 400 D4IPs with DPP-IV inhibition structure and high hydrophobicity were identified.The correlation network map showed that Lactococcus,Bacillus,Lysobacter,Pelagivirga,Kocuria,Escherichia,Streptococcus,and Peptostreptococcus were significantly correlated with the most D4IPs.Four stable D4IPs,including KAGARALTDAETAT,GEKVDFDDIQK,VVDADEMYLKGK,and GQKDSYVGDEAQ were respectively from the precursor proteins parvalbumin,troponin,myosin,and actin,and were mainly formed by the hydrolysis of subtilisin(EC 3.4.21.62),aspartic proteinase(EC 3.4.23.1),thermolysin(EC 3.4.24.27),oligopeptidase B(EC 3.4.21.83),and proteinase P1(EC 3.4.21.96)from Bacillus,Kocuria,Lysobacter,Lactococcus,and Peptostreptococcus.The inhibition mainly resulted from the hydrogen bond and salt bridge between D4IPs and DPP-IV enzyme.This study provides important information on the proteases and related microbial strains to directionally prepare D4IPs in Chouguiyu.

A nuclear import inhibitory peptide ameliorates the severity of cholecystokinin-induced acute pancreatitis

AIM: To assess the effect of our novel cell-permeable nuclear factor-kappaB (NF-κB) inhibitor peptide PN50 in an experimental model of acute pancreatitis. PN50 was produced by conjugating the cell-penetrating penetratin peptide with the nuclear localization signal of the NF-κB p50 subunit.METHODS: Pancreatitis was induced in male Wistar rats by administering 2×100 μg/kg body weight of cholecystokininoctapeptide (CCK) intraperitoneally (IP) at an interval of 1 h. PN50-treated animals received 1 mg/kg of PN50 IP 30 min before or after the CCK injections. The animals were sacrificed 4 h after the first injection of CCK.RESULTS: All the examined laboratory (the pancreatic weight/body weight ratio, serum amylase activity,pancreatic levels of TNF-α and IL-6, degree of lipid peroxidation, reduced glutathione levels, NF-κB binding activity, pancreatic and lung myeloperoxidase activity) and morphological parameters of the disease were improved before and after treatment with the PN50 peptide.According to the histological findings, PN50 protected the animals against acute pancreatitis by favoring the induction of apoptotic, as opposed to necrotic acinar cell death associated with severe acute pancreatitis.CONCLUSION: Our study implies that reversible inhibitors of stress-responsive transcription factors like NF-κB might be clinically useful for the suppression of the severity of acute pancreatitis.

Characterization of Angiotensin-? Converting Enzyme Inhibiting Peptide from Venerupis philippinarum with Nano-Liquid Chromatography in Combination with Orbitrap Mass Spectrum Detection and Molecular Docking

The complexity and diversity of peptide mixture from protein hydrolysates make their characterization difficult. In this study, a method combining nano LC-MS/MS with molecular docking was applied to identifying and characterizing a peptide with angiotensin-? converting enzyme(ACE-I) inhibiting activity from Venerupis philippinarum hydrolysate. Firstly, ethanol supernatant of V. philippinarum hydrolysate was separated into active fractions with chromatographic methods such as ion-exchange chromatography and high performance liquid chromatography in combination. Then seven peptides from active fraction were identified according to the searching result of the MS/MS spectra against protein databases. Peptides were synthesized and subjected to ACE-Iinhibition assay. The peptide NTLTLIDTGIGMTK showed the highest potency with an IC_(50) of 5.75 μmol L^(-1). The molecular docking analysis showed that the ACE-I inhibiting peptide NTLTLIDTGIGMTK bond with residues Glu123, Glu403, Arg522, Glu376, Gln281 and Asn285 of ACE-I. Therefore, active peptides could be identified with the present method rather than the traditional purification and identification strategies. It may also be feasible to identify other food-derived peptides which target other enzymes and receptors with the method developed in this study.

ACE-I inhibitory peptide fractions from enzymatic hydrolysates of velvet bean (<i>Mucuna pruriens</i>)

The hydrolysis of velvet bean (Mucuna pruriens) protein in the presence of Alcalase?-Flavourzyme? and Pepsin-Pancreatin was investigated. The results showed that Alcalase?-Flavourzyme? (29.08%) sequential system catalyzed the hydrolysis most efficiently that Pepsin-Pancreatin (24.78%). In addition, the higher ACE-I inhibitory activity was achieved with the sequential system Alcalase?-Flavourzyme? (33.13%). Furthermore, the concentration of peptides employing an ultrafiltration (UF) system or their purification by gel filtration chromatography showed that the oligomeric peptides with lower molecular weight registered the highest ACE-I inhibitory activity. It has been demonstrated that Mucuna pruriens protein hydrolysates could serve as a source of peptides with ACE inhibitory activity and this activity can be attributed mainly to the mixture of short peptides in the hydrolysate.

PTEN inhibition and axon regeneration and neural repair

The intrinsic growth ability of all the neurons declines during development although some may grow better than others. Numerous intracellular signaling proteins and transcription factors have been shown to regulate the intrinsic growth capacity in mature neurons. Among them, PI3 kinase/Akt pathway is important for controlling axon elongation. As a negative regulator of this pathway, the tumor suppressor phosphatase and tensin homolog （PTEN） appears critical to con- trol the regenerative ability of young and adult neurons. This review will focus on recent research progress in axon regeneration and neural repair by PTEN inhibition and therapeutic potential of blocking this phosphatase for neurological disorders. Inhibition of PTEN by deletion in con- ditional knockout mice, knockdown by short-hairpin RNA, or blockade by pharmacological approaches, including administration of selective PTEN antagonist peptides, stimulates various degrees of axon regrowth in juvenile or adult rodents with central nervous system injuries. Im- portantly, post-injury PTEN suppression could enhance axonal growth and functional recovery in adult central nervous system after injury.

Identifi cation and characterization of a novel tetrapeptide from enzymatic hydrolysates of Baijiu byproduct

In order to prepare angiotensin I-converting enzyme(ACE)inhibitory peptides,distilled spent grains of Chinese strong-flavor Baijiu were hydrolyzed by alcalase followed by papain under optimized conditions.A superior ACE inhibitory peptide was separated and purifi ed by ultrafi ltration and high-performance liquid chromatography(HPLC),and its amino acid sequence was further identified as Gln-Gly-Val-Pro(QGVP)by electrospray mass spectrometry(ESI-MS).QGVP formed 6 hydrogen bonds with the active site of ACE,which is responsible for reducingα-helix structure content of ACE causing subsequent inactivation.M oreover,it showed no significant cytotoxicity toward human umbilical vein endothelial cells(HUVECs),a nd signifi cantly i nduced phosphorylation of endothelial nitric oxide synthase(p-e NOS)and decreased endothelin 1(END1)expression in angiotensin I(Ang I)-treated HUVECs,demonstrating the potential antihypertensive effect.The peptide QGVP hydrolyzed from distilled spent grain proteins of Chinese strong-fl avor Baijiu was expected to be used as a food ingredient to prevent or co-treat hypertension with other chemical drugs.

Effects of heat shock protein-antigen peptide complexes (HACs) in melanoma B16 cell line on transplanted tumor development in mice

目的 :建立黑色素瘤 B16细胞热休克蛋白 -抗原肽复合物 (HACs)的制备方法并测其抑瘤效应。方法 :应用 Sephacryl S- 2 0 0凝胶过滤制备 HAC粗提物 ,应用 SDS- PAGE纯化 HACs,并测其抑瘤效应。结果 :应用 SDS- PAGE纯化的 HAC6 0、 HAC75和 HAC97不同程度地降低肿瘤发生率 ,延迟肿瘤发生时间和减慢肿瘤生长速度。结论 :6 0 0 0 0～ 970 0 0

基于分子对接的三角帆蚌酪氨酸酶抑制肽筛选及活性评价

【目的】筛选和鉴定三角帆蚌(Hyriopsis cumingii)珍珠层蛋白酶解产物中的酪氨酸酶抑制肽,为美白肽的研究与开发提供依据。【方法】以三角帆蚌珍珠层粉为原料,基于其蛋白组成特性,采用物理改性和可控酶解技术制备珍珠层活性肽(Peptides from nacre powder,P-NP),通过测定其2,2′-联氮双(3-乙基苯并噻唑啉-6-磺酸)二铵盐(ABTS)和1,1-二苯基-2-苦基肼自由基(DPPH)的清除能力、酪氨酸酶抑制活性以及对紫外线B(UVB)辐照损伤人永生化角质形成细胞(HaCaT)的影响,探究其体外活性。采用液质联用技术(LC-MS/MS)和分子对接技术从P-NP中鉴定并筛选出三角帆蚌酪氨酸酶抑制肽,并进行固相合成和体外活性验证。【结果】P-NP的ABTS和DPPH自由基清除效果较佳,肽质量浓度为1.00、14.00 mg/mL时,两者清除率分别为41.58%和11.23%;P-NP的酪氨酸酶活性抑制能力良好,其半抑制浓度(IC_(50))值为7.51 mg/mL。P-NP质量浓度在83.22μg/mL时,显著提高UVB损伤HaCaT细胞的生长活力(P<0.05)。P-NP经LC-MS/MS鉴定后筛选出相对分子质量集中在500~1000的61条肽序列作为候选肽库,分子对接筛选出3条潜在的酪氨酸酶抑制肽,其主要通过氢键与酪氨酸酶活性中心及其以外的位点相结合,从而达到抑制其活性的作用。其中,Tyr-Pro-Asn-Pro-Tyr(YPNPY)对酪氨酸酶抑制作用最强,IC_(50)为(0.545±0.028)mmol/L,主要抑制类型为竞争型抑制。【结论】三角帆蚌珍珠层活性肽具有良好的清除自由基和酪氨酸酶抑制活性。

白果肽的二肽基肽酶-Ⅳ抑制活性及其抑制机理研究

食源性二肽基肽酶-Ⅳ(dipeptidyl peptidaseⅣ,DPP-Ⅳ)抑制肽可有效调节机体内糖代谢,降低机体血糖水平。为评估白果肽(phenylalanine-alanine-proline-serine-tryptophan,FAPSW和methionine-proline-glycine-proline-proline,MPGPP)的降糖潜能,采用体外试验测定其DPP-Ⅳ抑制活性,并进一步通过分子对接和分子动力学模拟揭示其抑制机理。结果表明,FAPSW和MPGPP具有良好的DPP-Ⅳ抑制活性,且MPGPP的DPP-Ⅳ抑制活性[IC_(50)=(0.158±0.009)g/L]强于FAPSW[IC_(50)=(2.540±0.126)g/L];分子对接结果表明,静电相互作用、氢键、疏水作用和范德华力在FAPSW和MPGPP抑制DPP-Ⅳ活性中起重要作用;分子动力学结果表明,FAPSW和MPGPP结合DPP-Ⅳ能形成稳定的复合物结构,且MPGPP-DPP-Ⅳ复合物的稳定性要强于FAPSW-DPP-Ⅳ。结合自由能结果表明,静电相互作用在FAPSW和MPGPP结合DPP-Ⅳ中起主导作用。研究结果可为FAPSW和MPGPP在功能食品领域的开发和利用提供一定的理论参考。

虾壳源多肽酶解条件的优化及其抗冻活性和作用机制的探究

目的优化从虾壳中提取多肽的酶解条件,并探究虾壳多肽的抗冻活性及作用机制。方法以小龙虾虾壳为原料,使用复合酶酶解提取多肽。在单因素实验的基础上,通过正交实验得到虾壳最佳酶解条件。以面包酵母菌为抗冻测试对象测试其抗冻能力。采用液相色谱-串联质谱法(liquid chromatography-mass spectrometry,LC-MS/MS)对虾壳多肽的组成进行鉴定,基于多肽的理化性质和序列,结合抗冻预测平台筛选抗冻肽(antifreeze peptides,AFPs)。使用分子动力学模拟探究了AFPs的作用机制。结果在酶解温度40℃,底物质量浓度20mg/mL,加酶量12000U,酶比0.33的最佳酶解条件下,多肽提取率达到了70%以上。虾壳多肽中含有1004种肽段,能明显提升酵母菌存活率。从具有抗冻活性且置信度靠前的20条多肽中进一步筛选出了潜在的抗冻肽AFP1、AFP2。AFP2和冰面之间形成的氢键数量为14,部分水分子同时与2个氨基酸残基形成氢键,充当水桥稳定多肽的构象,AFP2与冰之间的结合能为-105.08 kcal/mol,冰结合模拟效果理想。结论AFPs与冰结合并抑制冰晶生长可能归因于带电荷的亲水性氨基酸残基与水分子形成的氢键、范德华力等分子间作用力,可开发为有益的冷冻保护剂,用于冷冻食品加工,具备良好的应用前景。

深低温保存下高效抗冻多肽的合理设计和机理探讨

The development of effective antifreeze peptides to control ice growth has attracted a significant amount of attention yet still remains a great challenge.Here,we propose a novel design method based on in-depth investigation of repetitive motifs in various ice-binding proteins(IBPs)with evolution analysis.In this way,several peptides with notable antifreeze activity were developed.In particular,a designed antifreeze peptide named AVD exhibits ideal ice recrystallization inhibition(IRI),solubility,and biocompatibility,making it suitable for use as a cryoprotective agent(CPA).A mutation analysis and molecular dynamics(MD)simulations indicated that the Thr6 and Asn8 residues of the AVD peptide are fundamental to its ice-binding capacity,while the Ser18 residue can synergistically enhance their interaction with ice,revealing the antifreeze mechanism of AVD.Furthermore,to evaluate the cryoprotection potential of AVD,the peptide was successfully employed for the cryopreservation of various cells,which demonstrated significant post-freezing cell recovery.This work opens up a new avenue for designing antifreeze materials and provides peptide-based functional modules for synthetic biology.

史氏鲟鱼皮肽的提取、鉴定及其血管紧张素转化酶抑制活性

【目的】确定史氏鲟(Acipenser schrenckii)鱼皮肽最佳酶解条件,了解其血管紧张素转换酶(ACE)抑制活性,探究史氏鲟鱼皮在ACE抑制活性肽段的开发应用的可行性。【方法】参照国标方法测定鲟鱼皮基本组成成分;以水解度和ACE抑制率为指标,采用单因素实验对最适水解酶、加酶量、料液比、温度以及时间确定鲟鱼鱼皮肽最佳酶解条件;使用不同截留相对分子质量的超滤膜设备分离纯化鲟鱼皮肽,并比较其ACE抑制率;随后对最优肽组分进行紫外光谱、分子质量范围、肽序列测定和生物学活性预测。【结果与结论】鲟鱼皮干基主要由蛋白质和脂肪组成。确定鲟鱼皮肽最佳酶解条件为使用碱性蛋白酶、酶添加量6000 U/g,料液质量体积比1 g∶5 mL,温度60℃,酶解时间7 h。分离纯化后得到3种相对分子质量不同的鲟鱼皮肽,分别为ASPs-1、ASPs-2和ASPs-3,其中最优肽组分ASPs-1的分子质量范围为172.13~2135.02u,共鉴定获得28条综合得分大于100的肽段,分子质量集中在807.45~1952.96 u;其中GPGGPSGERGPPGPM、GPSGPGGPPGPR、SGPPGFPGSPGPKGE、GPPGKDGQPGHPGPIGPA、GPAGPRGPAGPA五条肽段预测具有ACE抑制功能,水溶性好,无生物学毒性,说明ASPs-1作为ACE抑制剂,有运用于生物体的潜力。

鹿角多肽调控SLC7A11/GPX4轴抑制地塞米松诱导的成骨细胞铁死亡

背景:激素性股骨头坏死与成骨细胞铁死亡密切相关,鹿角多肽可以通过抑制成骨细胞铁死亡,促进成骨细胞的存活与功能的建立,具有治疗激素性股骨头坏死的潜力,但其对成骨细胞铁死亡的调控机制尚未明确。目的:探究鹿角多肽抑制地塞米松诱导成骨细胞铁死亡的作用机制。方法:①采用不同浓度梯度的鹿角多肽与地塞米松干预MC3T3-E114细胞,CCK-8法检测细胞活性,确定鹿角多肽与地塞米松的作用浓度;②采用不同质量浓度梯度鹿角多肽干预被地塞米松(800μmol/L)处理后的MC3T3-E114细胞,分为空白对照组、地塞米松组、地塞米松+鹿角多肽组,采用CCK-8法计算不同质量浓度鹿角多肽对地塞米松(800μmol/L)干预MC3T3-E114细胞增殖的影响;③借助试剂盒检测空白对照组、地塞米松组、地塞米松+鹿角多肽组中超氧化物歧化酶活性及谷胱甘肽、丙二醛、脂质过氧化物、细胞铁、活性氧含量的变化,并应用Western blot检测谷胱甘肽过氧化物酶4、溶质载体家族7成员11蛋白的表达,验证鹿角多肽抑制铁死亡的通路。结果与结论:①CCK-8法检测细胞活性,结果确定后续实验选择以鹿角多肽(10 mg/mL)和地塞米松(800μmol/L)干预MC3T3-E114细胞处理24 h;②地塞米松干预后,丙二醛、脂质过氧化物、细胞铁、活性氧含量均升高(P<0.01),谷胱甘肽含量、超氧化物歧化酶活性均降低(P<0.01),谷胱甘肽过氧化物酶4、溶质载体家族7成员11蛋白表达降低(P<0.05-0.01);鹿角多肽干预后,上述各指标变化明显被逆转(P<0.05-0.01);③结果表明,鹿角多肽可能通过调控溶质载体家族7成员11/谷胱甘肽过氧化物酶4轴抑制成骨细胞铁死亡,发挥对激素性股骨头坏死的治疗作用。

响应面法优化南极磷虾酶法制备DPP-IV抑制肽工艺条件的研究

采用动物蛋白水解酶酶解制备南极磷虾二肽基肽酶-IV(DPP-IV)抑制肽,以DPP-IV抑制率和可溶性蛋白含量为主要指标,考察温度、反应时间、pH、酶添加量对南极磷虾的酶解效果。在单因素的基础上,依据中心组合实验设计原理,运用4因素3水平的响应面分析法,建立动物蛋白水解酶酶解南极磷虾制备DPP-IV抑制肽的二次多项式数学模型,并以DPP-IV抑制率为响应值作响应面。结果表明:南极磷虾酶法制备DPP-IV抑制肽的最佳工艺参数为酶解时间3.85 h、温度45.02℃、pH值7.58、酶添加量237.55 U/g原料;在此条件下,酶解液DPP-IV抑制率的预测值为64.82%,验证值为64.78%,优化方法可行。

泥鳅抗氧化活性肽的制备、鉴定及其生物活性

泥鳅是亚洲常见的淡水鱼,具有较高的营养和药用价值。本研究采用短小芽孢杆菌4SP5分泌的蛋白酶水解泥鳅肉糜,超滤获得不同分子质量的水解产物。其中,分子质量小于5 ku的组分的抗氧化活性最强。经G25色谱过滤纯化,LC-MS/MS测序和鉴定过滤组分,获得6种潜在的生物活性肽。其中寡肽AFRVPTP具有良好的DPPH自由基清除活性(IC5037μg/mL)、羟自由基清除活性、超氧阴离子清除活性(IC500.73 mg/mL)。此外,该寡肽还具有抑制α-葡萄糖苷酶的作用,IC50为15.86 mg/mL。经在线预测,该寡肽无毒性和致敏性。分子对接结果表明,AFRVPTP通过与α-葡萄糖苷酶催化域的关键氨基酸结合而发挥作用,其可与Keap1活性口袋中的关键氨基酸残基(Asn387、Arg415、Tyr334、Arg380、Gln530、Tyr525、Arg483和Ser508)相互作用,表明该寡肽具有调节体内抗氧化作用的潜力。研究结果为泥鳅的精深加工提供参考依据。

大河乌猪火腿中新型α-葡萄糖苷酶抑制肽的分离、鉴定及活性研究

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(IC_(50)值为1.42 mg/mL)。BIOPE P-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,研究旨在为大河乌猪火腿肽的进一步开发和利用提供理论参考。

Head-to-tail cyclization of a heptapeptide eliminates its cytotoxicity and significantly increases its inhibition effect on amyloid β-protein fibrillation and cytotoxicity

Amyloid-β （Aβ） protein aggregation is the main hallmark of Alzheimer＇s disease （AD）. Inhibition of Aft fibrillation is thus a promising therapeutic approach to the prevention and treatment of AD. Recently, we designed a heptapeptide inhibitor, LVFFARK （LK7）. LK7 shows a promising inhibitory capability on Aft fibrillation, but is prone to self-assembling and displays high cytotoxicity, which would hinder its practical application. Herein, we modified LK7 by a head-to-tail cyclization and obtained a cyclic LK7 （cLK7）. cLK7 exhibits a different self-assembly behavior from LK7, and has higher stability against proteolysis than LK7 and little cytotoxicity to SH-SY5Y cells. Thermodynamic analysis revealed that both LK7 and cLK7 could bind to Aβ40 by electrostatic interactions, hydrogen bonding and hydrophobic interactions, but the binding affinity of cLK7 for Afl40 （KD = 4.96 μmol/L） is six times higher than that of LK7 （KD = 32.2 μmol/L）. The strong binding enables cLK7 to stabilize the secondary structure of Aβ40 and potently inhibit its nucleation, fibrillation and cytotoxicity at extensive concentration range, whereas LK7 could only moderately inhibit Aβ40 fibrillation and cytotoxicity at low concentrations. The findings indicate that the peptide cyclization is a promising approach to enhance the performance of peptide-based amyloid inhibitors.

响应面法优化香榧降血糖肽制备工艺

以香榧饼粕为原料,α-葡萄糖苷酶抑制率和蛋白水解度为主要考察指标,通过单因素实验和响应面法优化香榧降血糖肽的制备工艺。结果表明,香榧降血糖肽的最优酶解工艺参数为:碱性蛋白酶添加量10440.0 U/g,酶解pH值10.0,酶解温度46.0℃,酶解时间5.2 h,此时酶解制得蛋白水解度为24.38%的香榧降血糖肽,α-葡萄糖苷酶抑制率为59.11%。

沙棘籽粕蛋白肽的稳定性及分离纯化

以沙棘籽粕蛋白为原料,酶解制备具有抑制胰脂肪酶活性的多肽,通过超滤、大孔树脂分离、超高效液相色谱-串联质谱和分子对接对沙棘籽粕蛋白肽(seabuckthorn seed peptides from alcalase hydrolysates,SSPA)进行分离纯化、结构鉴定和筛选。以猪胰脂肪酶(porcine pancreatic lipase,PPL)抑制率为指标,稳定性分析表明,SSPA具有热稳定性和pH值稳定性,适量Na^(+)和Mg^(2+)可显著提高SSPA的PPL抑制活性,SSPA在短期贮藏过程中不会因暴露于空气而影响PPL抑制活性。超高效液相色谱-串联质谱鉴定得到31种多肽,结合分子对接筛选得到6个肽段,小于3 kDa组分中上述寡肽含量分别为VR(2.90%)、FR(7.40%)、RDR(1.10%)、APYR(1.50%)、NLLHR(1.40%)和EEAASLR(1.10%)。固相合成测定其IC_(50)值分别为VR(371.07μg/mL)、FR(243.07μg/mL)、RDR(250.50μg/mL)、APYR(350.41μg/mL)、NLLHR(220.70μg/mL)、EEAASLR(510.55μg/mL)。分子对接表明以上多肽通过氢键、π-π堆积与PPL结合。Pearson相关性分析表明,分子对接结合能与SSPA的PPL抑制率之间存在显著正相关(R^(2)=0.865,P<0.05)。

富硒核桃粕蛋白降血压肽的酶解制备及硒含量分析

以富硒核桃为原料经提油后得到富硒核桃粕,以血管紧张素转化酶(ACE)抑制作为评估标志,经单因素影响试验和响应面优化富硒核桃粕蛋白ACE抑制肽的制备工艺,并将酶解物进行超滤分离、氨基酸组成分析和硒含量测定。结果显示,富硒核桃粕蛋白ACE抑制肽的最佳酶解温度为35℃,酶解时间为2.52 h,pH值7.5、底物浓度5.39%、加酶量0.33%,在此试验条件下进行的验证试验ACE抑制率为79.13%,与理论值为78.84%较相符。最优酶解物经3 ku超滤膜分离,发现<3 ku超滤组分在1 mg/mL浓度下ACE抑制活性高达77.78%。同时,富硒核桃粕蛋白ACE抑制肽酶解物中疏水性氨基酸占27.19%,硒元素在酶解物和<3 ku超滤组分中的含量分别为0.82 mg/kg和1.35 mg/kg。该研究为食物源性补硒产品和辅助降血压健康食品的研究与开发提供了理论依据。