蛋清源活性肽的结构鉴定及生物活性研究
|
摘要
食源性蛋白质的生理活性主要归功于蛋白质内部经裂解后释放出的片段，即为活性肽。活性肽可通过体外特异性酶或是体内胃肠道消化酶裂解方式制备获得，对人类健康有巨大贡献。我国禽蛋产量居世界之首，且蛋清中含有多种功能蛋白组分，包括卵清蛋白，卵转铁蛋白和半胱氨酸蛋白酶抑制剂等。蛋清蛋白的酶解产物具有多种生理活性，包括抑菌活性，抗黏活性，降血压活性和抗氧化活性等，突显蛋清蛋白活性肽对人类健康及疾病的预防和治疗方面的巨大潜力。
本文通过色谱及组学技术分离纯化共获得19个活性肽片段，利用串联质谱鉴定19个活性肽的氨基酸序列，采用圆二色谱和傅立叶红外光谱探索其二级结构中α-螺旋，β-转角，β-折叠和无规卷曲等二级结构信息。研究分别对19个蛋清源活性肽的血管紧张素转化酶抑制活性、α葡萄糖苷酶抑制活性、α淀粉酶抑制活性和抗凝血酶活性进行了测定，筛选具有较高体外活性的活性肽，通过单次、短期和长期灌胃大鼠实验进行体内活性评价，借鉴基因组学技术分别从蛋白质水平和基因水平考察了蛋清源活性肽对大鼠体内目标蛋白和目标基因表达水平调控的影响，通过行为医学手段对长期服用活性肽大鼠的行为学进行了研究。
首先，通过对蛋清蛋白质进行生物酶解及分离纯化，经质谱碎裂离子比对结合软件解析蛋清源活性肽组分中共有19个活性肽，其氨基酸序列分别为RVPSLM, TPSPR, DLQGK, AGLAPY, RVPSL，DHPFLF, HAEIN，QIGLF，HANENIF，VKELY，TNGIIR，KLPGF，EAGVD，EVSGL，NVLQPS，QITKPN，LEPINF，AEAGVD和ANENIF。19个活性肽的分子量分布范围在500-850u之间，研究发现19个活性肽二级结构中α-螺旋、β-折叠和β-转角含量均在6%以下。
而后，建立血管紧张素转化酶抑制活性，α-淀粉酶抑制活性，α-葡萄糖苷酶抑制活性和凝血酶抑制活性的体外活性评价方法，筛选蛋清源活性肽的高活性组分。研究发现除了RVPSL和QIGLF具有较高血管紧张素转化酶抑制活性外，还发现了新的具有血管紧张素转化酶抑制活性的活性肽，其氨基酸序列为TNGIIR，半抑制浓度（IC50）为70μmol L-1；经α-淀粉酶抑制活性和α-葡萄糖苷酶抑制活性的测定，结果表明活性肽RVPSLM，TPSPR，KLPGF和NVLQPS具有较高的α-葡萄糖苷酶抑制活性，其对应的IC50分别为23.07μmol L-1，40.02μmol L-1,59.5μmol L-1和100.0μmol L-1，活性肽KLPGF具有α-淀粉酶抑制活性，其IC50为120.0±4.0μmol L-1；经凝血酶抑制活性筛选，表明RVPSL具有抗凝血酶活性开发的潜力。
通过行为医学技术评价活性肽RVPSL，QIGLF和TNGIIR肽对长期灌胃大鼠的神经系统，焦虑以及学习记忆等行为的影响。通过高架十字迷宫焦虑模型发现蛋清源活性肽RVPSL和TNGIIR没有降低大鼠在开放臂活动的次数和时间，而QIGLF和卡托普利则显著降低大鼠在开放臂活动的时间和次数。结合Morris水迷宫和避暗学习记忆模型发现蛋清源活性肽RVPSL，QIGLF和TNGIIR对大鼠的学习记忆的能力均无明显的降低作用。
最后，从大鼠的生理参数和蛋白表达水平以及基因表达水平三个方面，考察蛋清肽RVPSL，QIGLF和TNGIIR分别经单次灌胃，短期灌胃和长期灌胃原发性高血压大鼠后的作用效果。结果表明RVPSL和QIGLF的体内较低血压作用明显，并对大鼠的正常生长未造成影响，经长期灌胃RVPSL（50mg/kg bw）和QIGLF（50mg/kg bw）后大鼠的收缩压均降低30mmHg以上，而长期灌胃Captopril能够使大鼠的收缩压降低50mmHg；活性肽RVPSL和QIGLF对大鼠血清中Ang、Renin，AngⅡ和Aldosterone的蛋白质表达水平的影响结果表明活性肽RVPSL高剂量组原发性高血压大鼠血清中Renin，AngⅡ和Aldosterone的浓度及活性显著降低，可以证实活性肽RVPSL通过降低肾素活性和血管紧张素活性从而降低血管紧张素Ⅱ的生成量来实现调节血压的作用；通过探讨活性肽RVPSL对大鼠肾脏中目标基因表达的调节机制，表明RVPSL能够有效下调ACE和AT1表达水平，同时上调AT2的表达水平，同时也证明了AngⅡ的两个受体AT1和AT2在调节血压的作用上存在相反的生理功能。因此我们发现蛋清源活性肽RVPSL具有作为降血压功能成分的开发潜力。
本研究为提高禽蛋资源附加值及开发功能肽活性成分提供理论和应用参考，并对活性肽的作用机理提供了佐证。蛋清源活性肽的开发不仅具有较高的学术价值，而且将显著提高禽蛋附加值，延伸了禽蛋加工产业链，创造更大的经济效益。
Proteins are important components of foods and ingredients of food products. Inaddition to their nutritional role, proteins can act as functional food ingredients，including antihypertension, antioxidant, and anti-diabetic properties. Research carriedout to produce these foods has paid special attention to the study of the physiologicalrole played by dietary proteins. There are certain fragments within the sequence offood proteins that may show biological activity once released by hydrolysis. Thesefragments, known as bioactive peptides, can be produced in vivo by the action ofgastrointestinal enzymes and can also be obtained in vitro using specific enzymes, orduring the preparation of certain foods. Peptide research has contributed greatly tovarious areas of human health. The production of eggs in china is first in theworldwide. The egg white proteins include ovalbumin, ovotransferrin, ovomucoid,ovomucin, lysozyme, avidin, ovoinhibitor, ovomacroglobulin, and cystatin. Previousresearches suggested that hydrolysates of egg white proteins exhibited diverse ofphysiological properties, including novel antimicrobial activities, antiadhesiveproperties, immunomodulatory, anticancer, and antihypertensive activities, antioxidantproperties. Several biological activities have now been associated with hydrolysatesfrom egg white proteins, highlighting the importance of egg and egg components inhuman health and in disease prevention and treatment.
Following our previous work,19bioactive peptides have been isolated and purifiedby proteomics technology including chromatographic, tandem mass spectrometry.Subsequently, The secondary structures of those peptides, i.e., α-helix, β-angle,β-sheet, random coil, and other information, were performed using circular dichroism(CD) and Fourier transform infrared spectroscopy (FTIR). In vitro angiotensinconverting enzyme inhibitory activity, α-glucosidase inhibitory activity, α-amylaseinhibitory activity and anti-thrombin activity of those peptides from egg whiteproteins were measured, aimed to screen bioactive peptides with high activity. After the single oral administration, short-term oral administration and long-termadministration with peptides from egg white proteins for rats, target proteinsconcerntration from serum and the gene expression levels of kidney were evaluatedby methods of Real-Time PCR and Enzyme-linked immune assay. At the same time,behavior of rats administrated with peptides from egg white proteins was studied bythe behavioral medicine assay.
Firstly, egg white proteins were hydrolyzed and isolated, and the purified fractionof hydrolysates was characterized by tandem mass spectrometry. The result suggestedthat fraction of hydrolysates was identified and peptides sequence of the fraction wereRVPSLM, TPSPR, DLQGK, AGLAPY, RVPSL, DHPFLF, HAEIN, QIGLF,HANENIF, VKELY, TNGIIR, KLPGF, EAGVD, EVSGL, NVLQPS, QITKPN,LEPINF, AEAGVD and ANENIF, respectively. Those peptides consists of5-6aminoacid residues, and the molecular weight distribution of peptides range between500-850Da. Our result showed that α-helical, β-folding and β-angle components ofthose peptides were below6%by the Fourier transform infrared spectroscopy andcircular dichroism analysis.
Subsequently, in vitro angiotensin converting enzyme inhibitory activity, α-amylaseinhibitory activity, α-glucosidase inhibitory activity and inhibiting the activity againstthrombin activity of thsoes peptides were performed aimed to obtain high activitypeptides. Research have shown that QIGLF and RVPSL exhibited high activityagainst angiotensin converting enzyme, and found a novel angiotensin convertingenzyme inhibitory peptides, i.e., TNGIIR, with IC5070μmol/L; α-amylase inhibitoryactivity and α-glucosidase inhibitory activity of those peptides were measured, resultsshowed that bioactive peptide RVPSLM, TPSPR, KLPGF and NVLQPS performedhigher α-glucosidase inhibitory activity, and their values of IC50responsed to23.07μmol/L,40.02μmol/L,59.5μmol/L and100.0μmol/L, respectively. Peptide KLPGFshowed high α-amylase inhibitory activity with an IC50of120.0±4.0μmol/L. resultsalso suggested that peptide RVPSL was potential to exhibit high anti-thrombinactivity.
In addition, the present study was designed to evaluate the effect of RVPSL,QIGLF and TNGIIR orally administered on the anxiety-related behavior. Our workassessed the anxiolytic-like effects of those peptides after an oral administration in spontaneously hypertensive rats faced to the same behavioral situations using elevatedplus maze. The results showed that compared to negative control, the number ofentries into the open arms significantly decreased by QIGLF and Captopril treatment,but the number of entries did not decreased by TNGIIR and RVPSL treatment.Followed by the Morris water maze test and avoiding darkness test, the resultssuggested that egg whited derive peptides, i.e., RVPSL，QIGLF, and TNGIIR, did notaffect the ability of learning and memory for rats.
Finally, the research focused on the antihypertensive effect of peptides (i.e.,RVPSL，QIGLF, and TNGIIR) on mRNA Expression and physiological parameter ofspontaneously hypertensive rats (SHRs), after sigle oral administration, short-termadministration, and long-term administration, respectively. In addition, Kidney mRNAlevels of renin, ACE, and AT1and AT2receptors were detected by real-time PCR;besides, Ang I, Ang II, renin, and aldosterone concentrations of serum were alsomeasured. Our results indicated that blood pressure of treatment group administratedwith RVPSL (50mg/kg bw) and QIGLF (50mg/kg bw) to spontaneous hypertensiverats for long-term decreased significantly by30mmHg compared to the negativegroup, but which was still difference from that of captopril treatment group (withdecrease by50mmHg). At the same time, Serum Ang II, renin, and aldosteroneconcentrations of group treated with RVPSL were reduced compared to that of thenegative group. Subsequently, Kidney mRNA levels of renin, ACE, and AT1receptorof RVPSL treated group decreased significantly compared to that of negative group.our results indicated that the peptide RVPSL affected the expression of major RAScomponents by downregulating the ACE and AT1receptor while upregulating the AT2receptor in the kidney or serum of SHRs. Our findings suggested that RVPSL mightbe of potential use in the treatment or prevention of hypertension.
This study was able to investigate the in vivo bioactivity of the peptide from egg protein and was helpful to clarify the mechanism of bioactive peptides. The researchpossesses highly academic value and will enhance economic returns for enterpriseseventually.
本文通过色谱及组学技术分离纯化共获得19个活性肽片段，利用串联质谱鉴定19个活性肽的氨基酸序列，采用圆二色谱和傅立叶红外光谱探索其二级结构中α-螺旋，β-转角，β-折叠和无规卷曲等二级结构信息。研究分别对19个蛋清源活性肽的血管紧张素转化酶抑制活性、α葡萄糖苷酶抑制活性、α淀粉酶抑制活性和抗凝血酶活性进行了测定，筛选具有较高体外活性的活性肽，通过单次、短期和长期灌胃大鼠实验进行体内活性评价，借鉴基因组学技术分别从蛋白质水平和基因水平考察了蛋清源活性肽对大鼠体内目标蛋白和目标基因表达水平调控的影响，通过行为医学手段对长期服用活性肽大鼠的行为学进行了研究。
首先，通过对蛋清蛋白质进行生物酶解及分离纯化，经质谱碎裂离子比对结合软件解析蛋清源活性肽组分中共有19个活性肽，其氨基酸序列分别为RVPSLM, TPSPR, DLQGK, AGLAPY, RVPSL，DHPFLF, HAEIN，QIGLF，HANENIF，VKELY，TNGIIR，KLPGF，EAGVD，EVSGL，NVLQPS，QITKPN，LEPINF，AEAGVD和ANENIF。19个活性肽的分子量分布范围在500-850u之间，研究发现19个活性肽二级结构中α-螺旋、β-折叠和β-转角含量均在6%以下。
而后，建立血管紧张素转化酶抑制活性，α-淀粉酶抑制活性，α-葡萄糖苷酶抑制活性和凝血酶抑制活性的体外活性评价方法，筛选蛋清源活性肽的高活性组分。研究发现除了RVPSL和QIGLF具有较高血管紧张素转化酶抑制活性外，还发现了新的具有血管紧张素转化酶抑制活性的活性肽，其氨基酸序列为TNGIIR，半抑制浓度（IC50）为70μmol L-1；经α-淀粉酶抑制活性和α-葡萄糖苷酶抑制活性的测定，结果表明活性肽RVPSLM，TPSPR，KLPGF和NVLQPS具有较高的α-葡萄糖苷酶抑制活性，其对应的IC50分别为23.07μmol L-1，40.02μmol L-1,59.5μmol L-1和100.0μmol L-1，活性肽KLPGF具有α-淀粉酶抑制活性，其IC50为120.0±4.0μmol L-1；经凝血酶抑制活性筛选，表明RVPSL具有抗凝血酶活性开发的潜力。
通过行为医学技术评价活性肽RVPSL，QIGLF和TNGIIR肽对长期灌胃大鼠的神经系统，焦虑以及学习记忆等行为的影响。通过高架十字迷宫焦虑模型发现蛋清源活性肽RVPSL和TNGIIR没有降低大鼠在开放臂活动的次数和时间，而QIGLF和卡托普利则显著降低大鼠在开放臂活动的时间和次数。结合Morris水迷宫和避暗学习记忆模型发现蛋清源活性肽RVPSL，QIGLF和TNGIIR对大鼠的学习记忆的能力均无明显的降低作用。
最后，从大鼠的生理参数和蛋白表达水平以及基因表达水平三个方面，考察蛋清肽RVPSL，QIGLF和TNGIIR分别经单次灌胃，短期灌胃和长期灌胃原发性高血压大鼠后的作用效果。结果表明RVPSL和QIGLF的体内较低血压作用明显，并对大鼠的正常生长未造成影响，经长期灌胃RVPSL（50mg/kg bw）和QIGLF（50mg/kg bw）后大鼠的收缩压均降低30mmHg以上，而长期灌胃Captopril能够使大鼠的收缩压降低50mmHg；活性肽RVPSL和QIGLF对大鼠血清中Ang、Renin，AngⅡ和Aldosterone的蛋白质表达水平的影响结果表明活性肽RVPSL高剂量组原发性高血压大鼠血清中Renin，AngⅡ和Aldosterone的浓度及活性显著降低，可以证实活性肽RVPSL通过降低肾素活性和血管紧张素活性从而降低血管紧张素Ⅱ的生成量来实现调节血压的作用；通过探讨活性肽RVPSL对大鼠肾脏中目标基因表达的调节机制，表明RVPSL能够有效下调ACE和AT1表达水平，同时上调AT2的表达水平，同时也证明了AngⅡ的两个受体AT1和AT2在调节血压的作用上存在相反的生理功能。因此我们发现蛋清源活性肽RVPSL具有作为降血压功能成分的开发潜力。
本研究为提高禽蛋资源附加值及开发功能肽活性成分提供理论和应用参考，并对活性肽的作用机理提供了佐证。蛋清源活性肽的开发不仅具有较高的学术价值，而且将显著提高禽蛋附加值，延伸了禽蛋加工产业链，创造更大的经济效益。
Proteins are important components of foods and ingredients of food products. Inaddition to their nutritional role, proteins can act as functional food ingredients，including antihypertension, antioxidant, and anti-diabetic properties. Research carriedout to produce these foods has paid special attention to the study of the physiologicalrole played by dietary proteins. There are certain fragments within the sequence offood proteins that may show biological activity once released by hydrolysis. Thesefragments, known as bioactive peptides, can be produced in vivo by the action ofgastrointestinal enzymes and can also be obtained in vitro using specific enzymes, orduring the preparation of certain foods. Peptide research has contributed greatly tovarious areas of human health. The production of eggs in china is first in theworldwide. The egg white proteins include ovalbumin, ovotransferrin, ovomucoid,ovomucin, lysozyme, avidin, ovoinhibitor, ovomacroglobulin, and cystatin. Previousresearches suggested that hydrolysates of egg white proteins exhibited diverse ofphysiological properties, including novel antimicrobial activities, antiadhesiveproperties, immunomodulatory, anticancer, and antihypertensive activities, antioxidantproperties. Several biological activities have now been associated with hydrolysatesfrom egg white proteins, highlighting the importance of egg and egg components inhuman health and in disease prevention and treatment.
Following our previous work,19bioactive peptides have been isolated and purifiedby proteomics technology including chromatographic, tandem mass spectrometry.Subsequently, The secondary structures of those peptides, i.e., α-helix, β-angle,β-sheet, random coil, and other information, were performed using circular dichroism(CD) and Fourier transform infrared spectroscopy (FTIR). In vitro angiotensinconverting enzyme inhibitory activity, α-glucosidase inhibitory activity, α-amylaseinhibitory activity and anti-thrombin activity of those peptides from egg whiteproteins were measured, aimed to screen bioactive peptides with high activity. After the single oral administration, short-term oral administration and long-termadministration with peptides from egg white proteins for rats, target proteinsconcerntration from serum and the gene expression levels of kidney were evaluatedby methods of Real-Time PCR and Enzyme-linked immune assay. At the same time,behavior of rats administrated with peptides from egg white proteins was studied bythe behavioral medicine assay.
Firstly, egg white proteins were hydrolyzed and isolated, and the purified fractionof hydrolysates was characterized by tandem mass spectrometry. The result suggestedthat fraction of hydrolysates was identified and peptides sequence of the fraction wereRVPSLM, TPSPR, DLQGK, AGLAPY, RVPSL, DHPFLF, HAEIN, QIGLF,HANENIF, VKELY, TNGIIR, KLPGF, EAGVD, EVSGL, NVLQPS, QITKPN,LEPINF, AEAGVD and ANENIF, respectively. Those peptides consists of5-6aminoacid residues, and the molecular weight distribution of peptides range between500-850Da. Our result showed that α-helical, β-folding and β-angle components ofthose peptides were below6%by the Fourier transform infrared spectroscopy andcircular dichroism analysis.
Subsequently, in vitro angiotensin converting enzyme inhibitory activity, α-amylaseinhibitory activity, α-glucosidase inhibitory activity and inhibiting the activity againstthrombin activity of thsoes peptides were performed aimed to obtain high activitypeptides. Research have shown that QIGLF and RVPSL exhibited high activityagainst angiotensin converting enzyme, and found a novel angiotensin convertingenzyme inhibitory peptides, i.e., TNGIIR, with IC5070μmol/L; α-amylase inhibitoryactivity and α-glucosidase inhibitory activity of those peptides were measured, resultsshowed that bioactive peptide RVPSLM, TPSPR, KLPGF and NVLQPS performedhigher α-glucosidase inhibitory activity, and their values of IC50responsed to23.07μmol/L,40.02μmol/L,59.5μmol/L and100.0μmol/L, respectively. Peptide KLPGFshowed high α-amylase inhibitory activity with an IC50of120.0±4.0μmol/L. resultsalso suggested that peptide RVPSL was potential to exhibit high anti-thrombinactivity.
In addition, the present study was designed to evaluate the effect of RVPSL,QIGLF and TNGIIR orally administered on the anxiety-related behavior. Our workassessed the anxiolytic-like effects of those peptides after an oral administration in spontaneously hypertensive rats faced to the same behavioral situations using elevatedplus maze. The results showed that compared to negative control, the number ofentries into the open arms significantly decreased by QIGLF and Captopril treatment,but the number of entries did not decreased by TNGIIR and RVPSL treatment.Followed by the Morris water maze test and avoiding darkness test, the resultssuggested that egg whited derive peptides, i.e., RVPSL，QIGLF, and TNGIIR, did notaffect the ability of learning and memory for rats.
Finally, the research focused on the antihypertensive effect of peptides (i.e.,RVPSL，QIGLF, and TNGIIR) on mRNA Expression and physiological parameter ofspontaneously hypertensive rats (SHRs), after sigle oral administration, short-termadministration, and long-term administration, respectively. In addition, Kidney mRNAlevels of renin, ACE, and AT1and AT2receptors were detected by real-time PCR;besides, Ang I, Ang II, renin, and aldosterone concentrations of serum were alsomeasured. Our results indicated that blood pressure of treatment group administratedwith RVPSL (50mg/kg bw) and QIGLF (50mg/kg bw) to spontaneous hypertensiverats for long-term decreased significantly by30mmHg compared to the negativegroup, but which was still difference from that of captopril treatment group (withdecrease by50mmHg). At the same time, Serum Ang II, renin, and aldosteroneconcentrations of group treated with RVPSL were reduced compared to that of thenegative group. Subsequently, Kidney mRNA levels of renin, ACE, and AT1receptorof RVPSL treated group decreased significantly compared to that of negative group.our results indicated that the peptide RVPSL affected the expression of major RAScomponents by downregulating the ACE and AT1receptor while upregulating the AT2receptor in the kidney or serum of SHRs. Our findings suggested that RVPSL mightbe of potential use in the treatment or prevention of hypertension.
This study was able to investigate the in vivo bioactivity of the peptide from egg protein and was helpful to clarify the mechanism of bioactive peptides. The researchpossesses highly academic value and will enhance economic returns for enterpriseseventually.
引文
[1] Mine, Y.; D'Silva, I., Bioactive peptides in hen eggs [D]. Bioactive Natural Products:Detection, Isolation, and Structural Determination, Second Edition,2008,439-458.
[2] You, S. J.; Udenigwe, C. C.; Aluko, R. E.; Wu, J. P., Multifunctional peptides fromegg white lysozyme[J]. Food Research International,2010,43,848-855.
[3] Mine, Y., Recent advances in egg protein functionality in the food system[J]. WorldsPoultry Science Journal,2002,58,31-39.
[4] Nys, Y.; Sauveur, B., The nutritional value of eggs[J]. Productions Animales,2004,17,385-393.
[5] Fujita, H.; Usui, H.; Kurahashi, K.; Yoshikawa, M., Isolation and characterization ofovokinin, a bradykinin B1agonist peptide derived from ovalbumin[J]. Peptides,1995,16,785-90.
[6] Nisbet, A. D.; Saundry, R. H.; Moir, A. J. G.; Fothergill, L. A.; Fothergill, J. E., Thecomplete amino-acid-sequence of hen ovalbumin[J]. European Journal of Biochemistry,1981,115,335-345.
[7] Yang, M.; Mine, Y., Novel T-cell epitopes of ovalbumin in BALB/c mouse: Potentialfor peptide-immunotherapy[J]. Biochemical and Biophysical Research Communications,2009,378,203-208.
[8] Zambrowicz, A.; Zelazko, M.; Trziszka, T., Hen egg proteins: precursors of bioactivepeptides[J]. Medycyna Weterynaryjna,2011,67,157-161.
[9] Kurokawa, H.; Dewan, J. C.; Mikami, B.; Sacchettini, J. C.; Hirose, M., Crystalstructure of hen apo-ovotransferrin-Both lopes adopt an open conformation upon lossof iron[J]. Journal of Biological Chemistry,1999,274,28445-28452.
[10] Lee, N. K.; Ahn, D. U.; Park, K. K.; Paik, H. D., Inhibitory Effect onAngiotensin-converting Enzyme (ACE) and Optimization for Production ofOvotransferrin Hydrolysates[J]. Korean Journal for Food Science of Animal Resources,2010,30,286-290.
[11] Wu, J.; Acero-Lopez, A., Ovotransferrin: Structure, bioactivities, and preparation[J].Food Research International,2012,46,480-487.
[12] Moon, S. H.; Lee, J. H.; Lee, Y. J.; Paik, J. Y.; Ahn, D. U.; Paik, H. D., Antioxidant,Antimicrobial, and Cytotoxic Activities of Ovotransferrin from Egg White[J]. KoreanJournal for Food Science of Animal Resources,2012,32,612-617.
[13] Ferreira, M.; Oliveira, F. A. R.; Jost, R., Application of microfiltration to egg whitedepleted in ovomucin[J]. International Journal of Food Science and Technology,1999,34,27-32.
[14] Omana, D. A.; Wang, J. P.; Wu, J. P., Ovomucin-a glycoprotein with promisingpotential[J]. Trends in Food Science&Technology,2010,21,455-463.
[15] Offengenden, M.; Fentabil, M. A.; Wu, J. P., N-glycosylation of ovomucin fromhen egg white[J]. Glycoconjugate Journal,2011,28,113-123.
[16] Cegielska-Radziejewska, R.; Lesnierowski, G.; Szablewski, T.; Kijowski, J.,Physico-chemical properties and antibacterial activity of modified eggwhite-lysozyme[J]. European Food Research and Technology,2010,231,959-964.
[17] Jimenez-Saiz, R.; Martos, G.; Carrillo, W.; Lopez-Fandino, R.; Molina, E.,Susceptibility of lysozyme to in-vitro digestion and immunoreactivity of its digests[J].Food Chemistry,2011,127,1719-1726.
[18] Daeschel, M. A.; Zhao, Y., Lysozyme and its antimicrobial application in foods andbeverages[J]. Abstracts of Papers of the American Chemical Society,2009,237,67-67.
[19] Kerkaert, B.; Mestdagh, F.; De Meulenaer, B., Detection of hen's egg whitelysozyme in food: Comparison between a sensitive HPLC and a commercial ELISAmethod[J]. Food Chemistry,2010,120,580-584.
[20] Guimaraes, G. C.; Rojas, E. E. G.; Coelho, M. C.; Coimbra, J. S. R., Adsorptionkinetics and thermodynamic parameters of egg white proteins[J]. European FoodResearch and Technology,2011,232,985-993.
[21] Jolivet, P.; Boulard, C.; Beaumal, V.; Chardot, T.; Anton, M., Protein componentsof low-density lipoproteins purified from hen egg yolk[J]. Journal of Agricultural andFood Chemistry,2006,54,4424-4429.
[22] Jung, S.; Kim, D. H.; Son, J. H.; Nam, K.; Ahn, D. U.; Jo, C., The functionalproperty of egg yolk phosvitin as a melanogenesis inhibitor[J]. Food Chemistry,2012,135,993-998.
[23] Jiang, B.; Mine, Y., Preparation of novel functional oligophosphopeptides from henegg yolk phosvitin[J]. Journal of Agricultural and Food Chemistry,2000,48,990-994.
[24] Luo, Y.; Xiong, R. S.; Hou, L. F.; Li, J. D.; Chen, J. C., Process for Coproduction ofCrude Immunoglobulin Y and High-Density Lipoproteins from Hen Egg Yolk[J].Journal of Agricultural and Food Chemistry,2010,58,11420-11427.
[25] Castellani, O.; Belhomme, C.; David-Briand, E.; Guerin-Dubiard, C.; Anton, M.,Oil-in-water emulsion properties and interfacial characteristics of hen egg yolkphosvitin[J]. Food Hydrocolloids,2006,20,35-43.
[26] Samaraweera, H.; Zhang, W.-g.; Lee, E. J.; Ahn, D. U., Egg Yolk Phosvitin andFunctional Phosphopeptides-Review[J]. Journal of Food Science,2011,76, R143-R150.
[27] Zhang, X. W.; Qiu, N.; Geng, F.; Ma, M. H., Simply and effectively preparinghigh-purity phosvitin using polyethylene glycol and anion-exchange chromatography[J].Journal of Separation Science,2011,34,3295-3301.
[28]Segura-Campos, M.; Chel-Guerrero, L.; Betancur-Ancona, D.; Hernandez-Escalante,V. M., Bioavailability of Bioactive Peptides[J]. Food Reviews International,2011,27,213-226.
[29] Meisel, H., Food-derived bioactive proteins and peptides as potential componentsof nutraceuticals[J]. Current Pharmaceutical Design,2007,13,771-772.
[30] Xu, C. N.; Yang, C. B.; Yin, Y. G.; Liu, J. B.; Mine, Y., Phosphopeptides (PPPs)from hen egg yolk phosvitin exert anti-inflammatory activity via modulation of cytokineexpression[J]. Journal of Functional Foods,2012,4,718-726.
[31] Jakas, A. J.; Bjelis, N.; Piantanida, I.; Kralj, M., Synthesis and Biological Activityof Quercetin Derivatives of Endogenous Opiod Peptides Leu-and Met-enkephalin[J].Journal of Peptide Science,2010,16,123-123.
[32] Zakharov, N. D.; Legostaev, G. N.; Vokhmyakova, G. S., Changes inchemosensitive properties of central neurons induced by opiod peptides in helix-pomatiaand mice[J]. Journal of Evolutionary Biochemistry and Physiology,1986,22,117-119.
[33] Nagel, L.; Budke, C.; Dreyer, A.; Koop, T.; Sewald, N., Antifreeze glycopeptidediastereomers[J]. Beilstein Journal of Organic Chemistry,2012,8,1657-1667.
[34] Ebbinghaus, S.; Meister, K.; Prigozhin, M. B.; DeVries, A. L.; Havenith, M.;Dzubiella, J.; Gruebele, M., Functional Importance of Short-Range Binding andLong-Range Solvent Interactions in Helical Antifreeze Peptides[J]. Biophysical Journal,2012,103, L20-L22.
[35] Kovacs-Nolan, J.; Phillips, M.; Mine, Y., Advances in the value of eggs and eggcomponents for human health[J]. Journal of Agricultural and Food Chemistry,2005,53,8421-8431.
[36] Borges, B. C. D.; Souza, E. J.; da Costa, G. D. A.; Pinheiro, I. V. D.; Sinhoreti, M.A. C.; Braz, R.; Montes, M., Effect of dentin pre-treatment with a caseinphosphopeptide-amorphous calcium phosphate (CPP-ACP) paste on dentin bondstrength in tridimensional cavities[J]. Acta Odontologica Scandinavica,2013,71,271-277.
[37] Cheng, G.; Wang, Z. G.; Liu, Y. L.; Zhang, J. L.; Sun, D. H.; Ni, J. Z.,Phosphoprotein/Phosphopeptide Enrichment and Analysis Based on NanostructuredMaterials[J]. Progress in Chemistry,2013,25,620-632.
[38] Li, J. T.; Zhang, J. L.; He, H.; Ma, Z. L.; Nie, Z. K.; Wang, Z. Z.; Xu, X. G.,Apoptosis in human hepatoma HepG2cells induced by corn peptides and its anti-tumorefficacy in H22tumor bearing mice[J]. Food and Chemical Toxicology,2013,51,297-305.
[39] Ma, Z. L.; Zhang, W. J.; Yu, G. C.; He, H.; Zhang, Y., The primary structureidentification of a corn peptide facilitating alcohol metabolism by HPLC-MS/MS[J].Peptides,2012,37,138-143.
[40] Roblet, C.; Doyen, A.; Amiot, J.; Bazinet, L., Impact of pH on ultrafiltrationmembrane selectivity during electrodialysis with ultrafiltration membrane (EDUF)purification of soy peptides from a complex matrix[J]. Journal of Membrane Science,2013,435,207-217.
[41] Singh, A.; Banerjee, R., Peptide enriched functional food adjunct from soy whey: Astatistical optimization study[J]. Food Science and Biotechnology,2013,22,65-71.
[42] Tsou, M. J.; Kao, F. J.; Lu, H. C.; Kao, H. C.; Chiang, W. D., Purification andidentification of lipolysis-stimulating peptides derived from enzymatic hydrolysis of soyprotein[J]. Food Chemistry,2013,138,1454-1460.
[43] Nogata, Y.; Saito, T.; Abe, D.; Yoshida, A.; Ohta, H., Storage Stability of WheatBran Proteases for Producing Angiotensin I-converting Enzyme Inhibitory Peptides[J].Food Science and Technology Research,2013,19,313-317.
[44] Prandi, B.; Faccini, A.; Tedeschi, T.; Galaverna, G.; Sforza, S., LC/MS analysis ofproteolytic peptides in wheat extracts for determining the content of the allergenamylase/trypsin inhibitor CM3: Influence of growing area and variety[J]. FoodChemistry,2013,140,141-146.
[45] Campos-Quevedo, N.; Rosales-Mendoza, S.; Paz-Maldonado, L. M. T.;Martinez-Salgado, L.; Guevara-Arauza, J. C.; Soria-Guerra, R. E., Production ofmilk-derived bioactive peptides as precursor chimeric proteins in chloroplasts ofChlamydomonas reinhardtii[J]. Plant Cell Tissue and Organ Culture,2013,113,217-225.
[46] Fleminger, G.; Ragones, H.; Merin, U.; Silanikove, N.; Leitner, G., Low molecularmass peptides generated by hydrolysis of casein impair rennet coagulation of milk[J].International Dairy Journal,2013,30,74-78.
[47] Picariello, G.; Iacomino, G.; Mamone, G.; Ferranti, P.; Fierro, O.; Gianfrani, C.; DiLuccia, A.; Addeo, F., Transport across Caco-2monolayers of peptides arising from invitro digestion of bovine milk proteins[J]. Food Chemistry,2013,139,203-212.
[48] Pan, D. D.; Lu, H. Q.; Zeng, X. Q., A Newly Isolated Ca Binding Peptide fromWhey Protein[J]. International Journal of Food Properties,2013,16,1127-1134.
[49] Saiga, A.; Okumura, T.; Makihara, T.; Katsuta, S.; Shimizu, T.; Yamada, R.;Nishimura, T., Angiotensin I-converting enzyme inhibitory peptides in a hydrolyzedchicken breast muscle extract[J]. Journal of Agricultural and Food Chemistry,2003,51,1741-1745.
[50] Gomez-Ruiz, J. A.; Ramos, M.; Recio, I., Angiotensin-convertingenzyme-inhibitory peptides in Manchego cheeses manufactured with different startercultures[J]. International Dairy Journal,2002,12,697-706.
[51] Gimenez, B.; Aleman, A.; Montero, P.; Gomez-Guillen, M. C., Antioxidant andfunctional properties of gelatin hydrolysates obtained from skin of sole and squid[J].Food Chemistry,2009,114,976-983.
[52] Wang, S.; Agyare, K.; Damodaran, S., Optimisation of hydrolysis conditions andfractionation of peptide cryoprotectants from gelatin hydrolysate[J]. FoodChemistry,2009,115,620-630.
[53] Ranathunga, S.; Rajapakse, N.; Kim, S. K., Purification and characterization ofantioxidative peptide derived from muscle of conger eel (Conger myriaster)[J].European Food Research and Technology,2006,222,310-315.
[54] Sheih, I. C.; Fang, T. J.; Wu, T. K., Isolation and characterisation of a novelangiotensin I-converting enzyme (ACE) inhibitory peptide from the algae proteinwaste[J]. Food Chemistry,2009,115,279-284.
[55] Jung, W. K.; Jo, H. Y.; Qian, Z. J.; Jeong, Y. J.; Park, S. G.; Choi, I. W.; A novelanticoagulant protein with high affinity to blood coagulation factor Va from Tegillarcagranosa[J]. Journal of Biochemistry and Molecular Biology,2007,40,832-838.
[56] Sheih, I. C.; Wu, T. K.; Fang, T. J., Antioxidant properties of a new antioxidativepeptide from algae protein waste hydrolysate in different oxidation systems[J].Bioresource Technology,2009,100,3419-3425.
[57] Lee, J. K.; Hong, S.; Jeon, J. K.; Kim, S. K.; Byun, H. G., Purification andcharacterization of angiotensin I converting enzyme inhibitory peptides from the rotifer,Brachionus rotundiformis[J]. Bioresource Technology2009,100,5255-5259.
[58] Kim, S. Y.; Je, J. Y.; Kim, S. K., Purification and characterization of antioxidantpeptide from hoki (Johnius belengerii) frame protein by gastrointestinal digestion[J].Journal of Nutritional Biochemistry,2007,18,31-38.
[59] Lee, S. H.; Qian, Z. J.; Kim, S. K., A novel angiotensin I converting enzymeinhibitory peptide from tuna frame protein hydrolysate and its antihypertensive effect inspontaneously hypertensive rats[J]. Food Chemistry,2010,118,96-102.
[60] Jun, S. Y.; Park, P. J.; Jung, W. K.; Kim, S. K., Purification and characterization ofan antioxidative peptide from enzymatic hydrolysate of yellowfin sole (Limanda aspera)frame protein[J]. European Food Research and Technology,2004,219,20-26.
[61] Jung, W. K.; Karawita, R.; Heo, S. J.; Lee, B. J.; Kim, S. K.; Jeon, Y. J., Recoveryof a novel Ca-binding peptide from Alaska Pollack (Theragra chalcogramma) backboneby pepsinolytic hydrolysis[J]. Process Biochemistry,2006,41,2097-2100.
[62] Wilson, J.; Hayes, M.; Carney, B., Angiotensin-I-converting enzyme and prolylendopeptidase inhibitory peptides from natural sources with a focus on marineprocessing by-products[J]. Food Chemistry,2011,129,235-244.
[63] Lin, K. H.; Jaw, C. G.; Yen, J. H.; Wang, Y. S., Molecular connectivity indices forpredicting bioactivities of substituted nitrobenzene and aniline compounds[J].Ecotoxicology and Environmental Safety,2009,72,1942-1949.
[64] Liu, Y.; Ke, Z.; Cui, J.; Chen, W.-H.; Ma, L.; Wang, B., Synthesis, inhibitoryactivities, and QSAR study of xanthone derivatives as alpha-glucosidase inhibitors[J].Bioorganic&Medicinal Chemistry,2008,16,7185-7192.
[65] Majumder, K.; Wu, J. P., A new approach for identification of novelantihypertensive peptides from egg proteins by QSAR and bioinformatics[J]. FoodResearch International,2010,43,1371-1378.
[66] Pripp, A. H., Docking and virtual screening of ACE inhibitory dipeptides[J].European Food Research and Technology,2007,225,589-592.
[67] Pripp, A. H.; Isaksson, T.; Stepaniak, L.; Sorhaug, T., Quantitative structure-activityrelationship modelling of ACE-inhibitory peptides derived from milk proteins[J].European Food Research and Technology2004,219,579-583.
[68] Pripp, A. H.; Isaksson, T.; Stepaniak, L.; Sorhaug, T.; Ardo, Y., Quantitativestructure activity relationship modelling of peptides and proteins as a tool in foodscience[J]. Trends in Food Science&Technology,2005,16,484-494.
[69] Maryam, P.; Davood, A.; Niusha, S., Tertiary structure prediction ofalpha-Glucosidase and inhibition properties of N-(phenoxydecyl)phthalimidederivatives by modeling, Docking and QSAR[J]. Clinical Biochemistry2011,44,S101-S102.
[70] Wu, J. P.; Aluko, R. E.; Nakai, S., Structural requirements of angiotensinI-converting enzyme inhibitory peptides: Quantitative structure-activity relationshipmodeling of peptides containing4-10amino acid residues[J]. Qsar&CombinatorialScience,2006,25,873-880.
[71] Choi, I.; Jung, C.; Choi, H.; Kim, C.; Ha, H., Effectiveness of phosvitin peptides onenhancing bioavailability of calcium and its accumulation in bones[J]. Food Chemistry,2005,93,577-583.
[72] Mine, Y., Egg proteins and peptides in human health-chemistry, bioactivity andproduction[J]. Current Pharmaceutical Design,2007,13,875-884.
[73] Xu, C. N.; Yang, C. B.; Yin, Y. G.; Liu, J. B.; Mine, Y., Phosphopeptides (PPPs)from hen egg yolk phosvitin exert anti-inflammatory activity via modulation of cytokineexpression[J]. Journal of Functional Foods,2012,4,718-726.
[74] Fujita, H.; Sasaki, R.; Yoshikawa, M., Potentiation of the antihypertensive activityof orally administered ovokinin, a vasorelaxing peptide derived from ovalbumin, byemulsification in egg phosphatidylcholine[J]. Bioscience Biotechnology andBiochemistry,1995,59,2344-2345.
[75] You, S. J.; Udenigwe, C. C.; Aluko, R. E.; Wu, J. P., Multifunctional peptides fromegg white lysozyme[J]. Food Research International,2010,43,848-855.
[76] Davalos, A.; Miguel, M.; Bartolome, B.; Lopez-Fandino, R., Antioxidant activity ofpeptides derived from egg white proteins by enzymatic hydrolysis[J]. Journal of FoodProtection,2004,67,1939-1944.
[77] Wang, J.; Liang, Y.; Omana, D. A.; Kav, N. N. V.; Wu, J., Proteomics Analysis ofEgg White Proteins from Different Egg Varieties[J]. Journal of Agricultural and FoodChemistry,2012,60,272-282.
[78] Wu, J. P.; Aluko, R. E.; Muir, A. D., Improved method for direct high-performanceliquid chromatography assay of angiotensin-converting enzyme-catalyzed reactions[J].Journal of Chromatography A,2002,950,125-130.
[79] Wu, J. P.; Aluko, R. E.; Muir, A. D., Purification of angiotensin I-convertingenzyme-inhibitory peptides from the enzymatic hydrolysate of defatted canola meal[J].Food Chemistry,2008,111,942-950.
[80] Wu, J.; Acero-Lopez, A., Ovotransferrin: Structure, bioactivities, and preparation[J].Food Research International,2012,46,480-487.
[81] Wu, J. P.; Aluko, R. E., Quantitative structure-activity relationship study of bitterdi-and tri-peptides including relationship with angiotensin I-converting enzymeinhibitory activity[J]. Journal of Peptide Science,2007,13,63-69.
[82] Wu, J. P.; Aluko, R. E.; Nakai, S., Structural requirements of angiotensinI-converting enzyme inhibitory peptides: Quantitative structure-activity relationshipstudy of di-and tripeptides[J]. Journal of Agricultural and Food Chemistry,2006,54,732-738.
[83] Lv, L.; Chi, Y. J., Improvement of functional properties of ovalbuminphosphorylated by dry-heating in the presence of pyrophosphate[J]. European FoodResearch and Technology,2012,235,981-987.
[84] Chen, C.; Chi, Y. J., Antioxidant, ace inhibitory activities and functional propertiesof egg white protein hydrolysate[J]. Journal of Food Biochemistry,2012,36,383-394.
[85] Chen, C.; Chi, Y. J.; Lv, L., Purification and identification of antioxidant peptidesfrom egg white protein hydrolysate[J]. Amino Acids,2012,43,457-466.
[86] Liu, J. B.; Yu, Z. P.; Zhao, W. Z.; Lin, S. Y.; Wang, E. L.; Zhang, Y.; Hao, H.; Chen,F., Isolation and identification of angiotensin-converting enzyme inhibitory peptidesfrom egg white protein hydrolysates[J]. Food Chemistry,2010,122,1159-1163.
[87] Yu, Z. P.; Zhao, W. Z.; Liu, J. B.; Lu, J.; Chen, F., QIGLF, a novel angiotensinI-converting enzyme-inhibitory peptide from egg white protein[J]. Journal of theScience of Food and Agriculture,2011,91,921-926.
[88] Yu, Z. P.; Liu, B. Q.; Zhao, W. Z.; Yin, Y. G.; Liu, J. B.; Chen, F., Primary andsecondary structure of novel ACE-inhibitory peptides from egg white protein[J]. FoodChemistry2012,133,315-322.
[89] Liu Lijun,Zhu Chuanhe,Zhao Zheng. Analyzing molecular weight distribution ofwhey protein hydrolysates[J]. Food and bioproducts processing,2008,86(C1):1-6.
[90]刘静波,于志鹏,赵文竹,等.蛋清源ACE抑制肽结构鉴定及其稳定性[J].吉林大学学报(工学版),2011,(02):579-584.
[91]于志鹏.蛋清源ACE抑制肽的结构鉴定及稳定性研究[D].长春:吉林大学,2011.
[92] Yu, Z.; Yin, Y.; Zhao, W.; Wang, F.; Yu, Y.; Liu, B.; Liu, J.; Chen, F.,Characterization of ACE-Inhibitory Peptide Associated with Antioxidant andAnticoagulation Properties[J]. Journal of Food Science,2011,76, C1149-C1155.
[93]张超,张晖,赵晓燕,马越,姚惠源.使用圆二色性光谱和红外光谱研究冬小麦麸皮抗冻蛋白的二级结构[J].光谱学与光谱分析,2009,29(7):1764-1767.
[94] Murariu, M.; Dragan, E. S.; Drochioiu, G., IR, MS and CD Investigations onSeveral Conformationally-Different Histidine Peptides[J]. International Journal ofPeptide Research and Therapeutics,2009,15,303-311.
[95] Carbonaro, M.; Nucara, A., Secondary structure of food proteins by Fouriertransform spectroscopy in the mid-infrared region[J]. Amino Acids,2010,38,679-690.
[96] Miguel, M.; Recio, I.; Gomez-Ruiz, J. A.; Ramos, M.; Lopez-Fandino, R.,Angiotensin I-converting enzyme inhibitory activity of peptides derived from egg whiteproteins by enzymatic hydrolysis[J]. Journal of Food Protection,2004,67,1914-1920.
[97] Marcuse, R., Antioxidative effect of amino-acids[J]. Nature,1960,186,886-887.
[98] Chen, C.; Chi, Y. J., antioxidant, ace inhibitory activities and functional propertiesof egg white protein hydrolysate[J].Journal of Food Biochemistry,2012,36(4),383-394.
[99]Chen, C.; Chi, Y. J.; Zhao, M. Y.; Lv, L., Purification and identification ofantioxidant peptides from egg white protein hydrolysate[J]. Amino Acids,2012,43(1),457-466.
[100] You, S. J.; Udenigwe, C. C.; Aluko, R. E.; Wu, J. P., Multifunctional peptides fromegg white lysozyme[J]. Food Research International,2010,43(3),848-855.
[101] Li, B.; Chen, F.; Wang, X.; Ji, B. P.; Wu, Y. N., Isolation and identification ofantioxidative peptides from porcine collagen hydrolysate by consecutivechromatography and electrospray ionization-mass spectrometry[J]. Food Chemistry,2007,102(4),1135-1143.
[102] Zhong, S.; Ma, C.; Lin, Y. C.; Luo, Y., Antioxidant properties of peptide fractionsfrom silver carp (Hypophthalmichthys molitrix) processing by-product proteinhydrolysates evaluated by electron spin resonance spectrometry[J]. Food Chemistry,2011,126(4),1636-1642.
[103]王嘉荣，滕达，田子罡，等.功能性抗氧化肽制备与机制研究进展[J].天然产物研究与开发,2008,20:371-375.
[104] Kokubu, T.; Ueda, E.; Fujimoto, S.; Hiwada, K.; Kato, A.; Akutsu, H.; Yamamura,Y.; Saito, S.; Mizoguch.T, Peptide inhibitors of renin-angiotensin system[J]. Nature,1968,217(5127),456-&.
[105] Ishiguro, K.; Sameshima, Y.; Kume, T.; Ikeda, K.-i.; Matsumoto, J.; Yoshimoto,M., Hypotensive effect of a sweetpotato protein digest in spontaneously hypertensiverats and purification of angiotensin I-converting enzyme inhibitory peptides[J]. FoodChemistry,2012,131(3),774-779.
[106] Guang, C.; Phillips, R. D.; Jiang, B.; Milani, F., Three key proteases-angiotensin-l-converting enzyme (ACE), ACE2and renin-within and beyond therenin-angiotensin system[J]. Archives of Cardiovascular Diseases,2012,105(6-7),373-385.
[107] Lin, L.; Lv, S.; Li, B., Angiotensin-I-converting enzyme (ACE)-inhibitory andantihypertensive properties of squid skin gelatin hydrolysates[J]. Food Chemistry,2012,131(1),225-230.
[108] Li, Y.; Zhou, J.; Huang, K.; Sun, Y.; Zeng, X., Purification of a Novel AngiotensinI-Converting Enzyme (ACE) Inhibitory Peptide with an Antihypertensive Effect fromLoach (Misgurnus anguillicaudatus)[J]. Journal of agricultural and food chemistry,2012,60(5),1320-5.
[109] Lu, J.; Sawano, Y.; Miyakawa, T.; Xue, Y.-L.; Cai, M.-Y.; Egashira, Y.; Ren, D.-F.;Tanokura, M., One-Week Antihypertensive Effect of Ile-Gln-Pro in SpontaneouslyHypertensive Rats[J]. Journal of Agricultural and Food Chemistry,2011,59(2),559-563.
[110] Wang, X.; Wang, L.; Cheng, X.; Zhou, J.; Tang, X.; Mao, X. Y.,Hypertension-attenuating effect of whey protein hydrolysate on spontaneouslyhypertensive rats[J]. Food Chemistry,2012,134(1),122-126.
[111] Li, Y.; Zhou, J.; Huang, K.; Sun, Y.; Zeng, X., Purification of a Novel AngiotensinI-Converting Enzyme (ACE) Inhibitory Peptide with an Antihypertensive Effect fromLoach (Misgurnus anguillicaudatus)[J]. Journal of Agricultural and Food Chemistry,2012,60(5),1320-1325.
[112] Ghosh, S.; Ahire, M.; Patil, S.; Jabgunde, A.; Bhat Dusane, M.; Joshi, B. N.;Pardesi, K.; Jachak, S.; Dhavale, D. D.; Chopade, B. A., Antidiabetic Activity of Gnidiaglauca and Dioscorea bulbifera: Potent Amylase and Glucosidase Inhibitors[J].Evidence-based complementary and alternative medicine: eCAM,2012,2012,929051.
[113] Lee, H. J.; Lee, H.-S.; Choi, J. W.; Ra, K. S.; Kim, J.-M.; Suh, H. J., NovelTripeptides with alpha-Glucosidase Inhibitory Activity Isolated from Silk CocoonHydrolysate[J]. Journal of agricultural and food chemistry,2011,59(21),11522-5.
[114] Kim, Y. M.; Wang, M. H.; Rhee, H. I., A novel alpha-glucosidase inhibitor frompine bark[J]. Carbohydrate Research,2004,339(3),715-717.
[115] Yu, Z. P.; Yin, Y. G.; Zhao, W. Z.; Yu, Y. D.; Liu, B. Q.; Liu, J. B.; Chen, F., Novelpeptides derived from egg white protein inhibiting alpha-glucosidase[J]. FoodChemistry,2011,129(4),1376-1382.
[116]杨万根.蛋清蛋白水解物的制备结构及其生物活性的研究[D].无锡，江南大学，2008.
[117] Yu, Z. P.; Yin, Y. G.; Zhao, W. Z.; Liu, J. B.; Chen, F., Anti-diabetic activitypeptides from albumin against alpha-glucosidase and alpha-amylase[J]. Food Chemistry,2012,135(3),2078-2085.
[118]王本祥，杨明，金玉莲，崔志勇，王岩.人参多肽降血糖作用[J].药学学报，1990，25(6):401-405.
[119]吴鸿,邹大进.糖尿病新药应用进展[J].中国实用内科杂志.2009,29(10):962-963.
[120]张瑞东,迟玉杰,陈晨,阮长青.蛋清蛋白酶解物抗氧化与血管紧张素转换酶抑制活性的研究[J].营养学报,2010,32(4).
[121]李亚云,赵新淮.两步法制备高活性酪蛋白ACE抑制肽的工艺参数研究[J].食品科学，31(10):6-11.
[122] Asoodeh, A.; Yazdi, M. M.; Chamani, J., Purification and characterisation ofangiotensin I converting enzyme inhibitory peptides from lysozyme hydrolysates[J].Food Chemistry,2012,131(1),291-295.
[123] Chen, C.; Chi, Y. J., Antioxidant, ace inhibitory activities and functional propertiesof egg white protein hydrolysate[J]. Journal of Food Biochemistry,2012,36(4),383-394.
[124] Garcia-Redondo, A. B.; Roque, F. R.; Miguel, M.; Lopez-Fandino, R.; Salaices,M., Vascular effects of egg white-derived peptides in resistance arteries from rats.Structure-activity relationships[J]. Journal of the Science of Food and Agriculture,2010,90(12),1988-1993.
[125] Miguel, M.; Recio, I.; Gomez-Ruiz, J. A.; Ramos, M.; Lopez-Fandino, R.,Angiotensin I-converting enzyme inhibitory activity of peptides derived from egg whiteproteins by enzymatic hydrolysis[J]. Journal of Food Protection,2004,67(9),1914-1920.
[126] Miguel, M.; Lopez-Fandino, R.; Ramos, M.; Aleixandre, A., Short-term effect ofegg-white hydrolysate products on the arterial blood pressure of hypertensive rats[J].British Journal of Nutrition,2005,94(5),731-737.
[127] Miguel, M.; Lopez-Fandino, R.; Ramos, M.; Aleixandre, A., Long-term intake ofegg white hydrolysate attenuates the development of hypertension in spontaneouslyhypertensive rats[J]. Life Sciences,2006,78(25),2960-2966.
[128]蒋菁莉,任发政.食源性降血压肽的评价方法[J].中国乳品工业,2006，34(6):36-39.
[129]卢静.大豆肽对大鼠的心血管效应及机制研究[D].长春：吉林大学，2006.
[130] Wang, X.; Wang, L.; Cheng, X.; Zhou, J.; Tang, X.; Mao, X. Y.,Hypertension-attenuating effect of whey protein hydrolysate on spontaneouslyhypertensive rats[J]. Food Chemistry2012,134(1),122-126.
[131]鲁军.螺旋藻活性肽的纯化鉴定及降压护肝机制研究[D].北京：北京林业大学，2009.
[132] He, R.; Alashi, A.; Malomo, S. A.; Girgih, A. T.; Chao, D. F.; Ju, X. R.; Aluko, R.E., Antihypertensive and free radical scavenging properties of enzymatic rapeseedprotein hydrolysates[J]. Food Chemistry2013,141(1),153-159.
[133] Fujita, H.; Yokoyama, K.; Yoshikawa, M., Classification and antihypertensiveactivity of angiotensin I-converting enzyme inhibitory peptides derived from foodproteins[J]. Journal of Food Science2000,65(4),564-569.
[134] Fujita, H.; Sasaki, R.; Yoshikawa, M., Potentiation of the antihypertensive activityof orally administered ovokinin, a vasorelaxing peptide derived from ovalbumin, byemulsification in egg phosphatidylcholine[J]. Bioscience Biotechnology andBiochemistry1995,59(12),2344-2345.
[135] Miguel, M.; Alvarez, Y.; Lopez-Fandino, R.; Alonso, M. J.; Salaices, M.,Vasodilator effects of peptides derived from egg white proteins[J]. Regulatory Peptides2007,140(3),131-135.
[136] Miguel, M.; Lopez-Fandino, R.; Aleixandre, M. A., Effect of the hydrolysateobtained from egg white with pepsin on the development of hypertension ofspontaneously hypertensive rats[J]. Ciencia y Tecnologia Alimentaria2005,4(5),368-372.
[137]陈季旺,刘珊珊,蔡广霞,吴永宁.食物源血管紧张素转化酶抑制三肽的定量构效关系[J].食品科学,2013,34（9）：19-23.
[138] Kearney, P. M.; Whelton, M.; Reynolds, K.; Muntner, P.; Whelton, P. K.; He, J.,Global burden of hypertension: analysis of worldwide data[J]. Lancet2005,365(9455),217-223.
[139] Violle, N.; Messaoudi, M.; Lefranc-Millot, C.; Desor, D.; Nejdi, A.; Demagny, B.;Schroeder, H., Ethological comparison of the effects of a bovine alpha(s1)-casein tryptichydrolysate and diazepam on the behaviour of rats in two models of anxiety[J].Pharmacology Biochemistry and Behavior2006,84(3),517-523;
[140] Walf, A. A.; Frye, C. A., The use of the elevated plus maze as an assay ofanxiety-related behavior in rodents[J]. Nature Protocols2007,2(2),322-328.
[141]刘爽,夏杨毅,尚永彪.鸡蛋蛋清蛋白酶解产物的研究进展[J].食品科学,2013,34(15):409-414.
[142] Hooge, R.; De Deyn, P. P., Applications of the Morris water maze in the study oflearning and memory[J]. Brain Research Reviews2001,36(1),60-90.
[143] McNamara, R. K.; Skelton, R. W., The neuropharmacological and neurochemicalbasis of place learning in the morris water maze[J]. Brain Research Reviews1993,18(1),33-49.
[144] Capozziello, S.; De Laurentis, M.; Nojiri, S.; Odintsov, S. D., Classifying andavoiding singularities in the alternative gravity dark energy models[J]. Physical ReviewD2009,79(12).
[145] Gilbert, P.; McEwan, K.; Bellew, R.; Mills, A.; Gale, C., The dark side ofcompetition: How competitive behaviour and striving to avoid inferiority are linked todepression, anxiety, stress and self-harm[J]. Psychology and Psychotherapy-TheoryResearch and Practice2009,82(2),123-136.
[146] Schijven, P.; Mulken, O., Avoiding dark states in open quantum systems bytailored initializations[J]. Physical Review E2012,85(6).
[147]许崇涛,李慧,林凌云.快眼动睡眠剥夺对抑郁模型大鼠自主活动及海马5-羟色胺含量的影响[J].中国行为医学科学,2004,13(4):381-383.
[148]刘屏,陈世铭,侯津详.高架十字迷宫和Vogel饮水冲突法用于吗啡依赖动物焦虑评价[J].中国药理学报,1997,13(6):563-565.
[149]宗绍波,魏盛,苏云祥,乔明琦.焦虑大鼠模型高架十字迷宫实验的复测信度检验及参数相关性分析[J].中国医药导报,2011,8(30):5-7.
[150]陈罗西,郭玲玲,李亮. Morris圆形水迷宫的应用及相关检测指标分析[J].辽宁中医药大学学报,2008,10(8):55-57.
[151]胡镜清,温泽淮,赖世隆. Morris水迷宫检测的记忆属性与方法学初探[J].广州中医药大学学报,2000,17(2):117-119.
[2] You, S. J.; Udenigwe, C. C.; Aluko, R. E.; Wu, J. P., Multifunctional peptides fromegg white lysozyme[J]. Food Research International,2010,43,848-855.
[3] Mine, Y., Recent advances in egg protein functionality in the food system[J]. WorldsPoultry Science Journal,2002,58,31-39.
[4] Nys, Y.; Sauveur, B., The nutritional value of eggs[J]. Productions Animales,2004,17,385-393.
[5] Fujita, H.; Usui, H.; Kurahashi, K.; Yoshikawa, M., Isolation and characterization ofovokinin, a bradykinin B1agonist peptide derived from ovalbumin[J]. Peptides,1995,16,785-90.
[6] Nisbet, A. D.; Saundry, R. H.; Moir, A. J. G.; Fothergill, L. A.; Fothergill, J. E., Thecomplete amino-acid-sequence of hen ovalbumin[J]. European Journal of Biochemistry,1981,115,335-345.
[7] Yang, M.; Mine, Y., Novel T-cell epitopes of ovalbumin in BALB/c mouse: Potentialfor peptide-immunotherapy[J]. Biochemical and Biophysical Research Communications,2009,378,203-208.
[8] Zambrowicz, A.; Zelazko, M.; Trziszka, T., Hen egg proteins: precursors of bioactivepeptides[J]. Medycyna Weterynaryjna,2011,67,157-161.
[9] Kurokawa, H.; Dewan, J. C.; Mikami, B.; Sacchettini, J. C.; Hirose, M., Crystalstructure of hen apo-ovotransferrin-Both lopes adopt an open conformation upon lossof iron[J]. Journal of Biological Chemistry,1999,274,28445-28452.
[10] Lee, N. K.; Ahn, D. U.; Park, K. K.; Paik, H. D., Inhibitory Effect onAngiotensin-converting Enzyme (ACE) and Optimization for Production ofOvotransferrin Hydrolysates[J]. Korean Journal for Food Science of Animal Resources,2010,30,286-290.
[11] Wu, J.; Acero-Lopez, A., Ovotransferrin: Structure, bioactivities, and preparation[J].Food Research International,2012,46,480-487.
[12] Moon, S. H.; Lee, J. H.; Lee, Y. J.; Paik, J. Y.; Ahn, D. U.; Paik, H. D., Antioxidant,Antimicrobial, and Cytotoxic Activities of Ovotransferrin from Egg White[J]. KoreanJournal for Food Science of Animal Resources,2012,32,612-617.
[13] Ferreira, M.; Oliveira, F. A. R.; Jost, R., Application of microfiltration to egg whitedepleted in ovomucin[J]. International Journal of Food Science and Technology,1999,34,27-32.
[14] Omana, D. A.; Wang, J. P.; Wu, J. P., Ovomucin-a glycoprotein with promisingpotential[J]. Trends in Food Science&Technology,2010,21,455-463.
[15] Offengenden, M.; Fentabil, M. A.; Wu, J. P., N-glycosylation of ovomucin fromhen egg white[J]. Glycoconjugate Journal,2011,28,113-123.
[16] Cegielska-Radziejewska, R.; Lesnierowski, G.; Szablewski, T.; Kijowski, J.,Physico-chemical properties and antibacterial activity of modified eggwhite-lysozyme[J]. European Food Research and Technology,2010,231,959-964.
[17] Jimenez-Saiz, R.; Martos, G.; Carrillo, W.; Lopez-Fandino, R.; Molina, E.,Susceptibility of lysozyme to in-vitro digestion and immunoreactivity of its digests[J].Food Chemistry,2011,127,1719-1726.
[18] Daeschel, M. A.; Zhao, Y., Lysozyme and its antimicrobial application in foods andbeverages[J]. Abstracts of Papers of the American Chemical Society,2009,237,67-67.
[19] Kerkaert, B.; Mestdagh, F.; De Meulenaer, B., Detection of hen's egg whitelysozyme in food: Comparison between a sensitive HPLC and a commercial ELISAmethod[J]. Food Chemistry,2010,120,580-584.
[20] Guimaraes, G. C.; Rojas, E. E. G.; Coelho, M. C.; Coimbra, J. S. R., Adsorptionkinetics and thermodynamic parameters of egg white proteins[J]. European FoodResearch and Technology,2011,232,985-993.
[21] Jolivet, P.; Boulard, C.; Beaumal, V.; Chardot, T.; Anton, M., Protein componentsof low-density lipoproteins purified from hen egg yolk[J]. Journal of Agricultural andFood Chemistry,2006,54,4424-4429.
[22] Jung, S.; Kim, D. H.; Son, J. H.; Nam, K.; Ahn, D. U.; Jo, C., The functionalproperty of egg yolk phosvitin as a melanogenesis inhibitor[J]. Food Chemistry,2012,135,993-998.
[23] Jiang, B.; Mine, Y., Preparation of novel functional oligophosphopeptides from henegg yolk phosvitin[J]. Journal of Agricultural and Food Chemistry,2000,48,990-994.
[24] Luo, Y.; Xiong, R. S.; Hou, L. F.; Li, J. D.; Chen, J. C., Process for Coproduction ofCrude Immunoglobulin Y and High-Density Lipoproteins from Hen Egg Yolk[J].Journal of Agricultural and Food Chemistry,2010,58,11420-11427.
[25] Castellani, O.; Belhomme, C.; David-Briand, E.; Guerin-Dubiard, C.; Anton, M.,Oil-in-water emulsion properties and interfacial characteristics of hen egg yolkphosvitin[J]. Food Hydrocolloids,2006,20,35-43.
[26] Samaraweera, H.; Zhang, W.-g.; Lee, E. J.; Ahn, D. U., Egg Yolk Phosvitin andFunctional Phosphopeptides-Review[J]. Journal of Food Science,2011,76, R143-R150.
[27] Zhang, X. W.; Qiu, N.; Geng, F.; Ma, M. H., Simply and effectively preparinghigh-purity phosvitin using polyethylene glycol and anion-exchange chromatography[J].Journal of Separation Science,2011,34,3295-3301.
[28]Segura-Campos, M.; Chel-Guerrero, L.; Betancur-Ancona, D.; Hernandez-Escalante,V. M., Bioavailability of Bioactive Peptides[J]. Food Reviews International,2011,27,213-226.
[29] Meisel, H., Food-derived bioactive proteins and peptides as potential componentsof nutraceuticals[J]. Current Pharmaceutical Design,2007,13,771-772.
[30] Xu, C. N.; Yang, C. B.; Yin, Y. G.; Liu, J. B.; Mine, Y., Phosphopeptides (PPPs)from hen egg yolk phosvitin exert anti-inflammatory activity via modulation of cytokineexpression[J]. Journal of Functional Foods,2012,4,718-726.
[31] Jakas, A. J.; Bjelis, N.; Piantanida, I.; Kralj, M., Synthesis and Biological Activityof Quercetin Derivatives of Endogenous Opiod Peptides Leu-and Met-enkephalin[J].Journal of Peptide Science,2010,16,123-123.
[32] Zakharov, N. D.; Legostaev, G. N.; Vokhmyakova, G. S., Changes inchemosensitive properties of central neurons induced by opiod peptides in helix-pomatiaand mice[J]. Journal of Evolutionary Biochemistry and Physiology,1986,22,117-119.
[33] Nagel, L.; Budke, C.; Dreyer, A.; Koop, T.; Sewald, N., Antifreeze glycopeptidediastereomers[J]. Beilstein Journal of Organic Chemistry,2012,8,1657-1667.
[34] Ebbinghaus, S.; Meister, K.; Prigozhin, M. B.; DeVries, A. L.; Havenith, M.;Dzubiella, J.; Gruebele, M., Functional Importance of Short-Range Binding andLong-Range Solvent Interactions in Helical Antifreeze Peptides[J]. Biophysical Journal,2012,103, L20-L22.
[35] Kovacs-Nolan, J.; Phillips, M.; Mine, Y., Advances in the value of eggs and eggcomponents for human health[J]. Journal of Agricultural and Food Chemistry,2005,53,8421-8431.
[36] Borges, B. C. D.; Souza, E. J.; da Costa, G. D. A.; Pinheiro, I. V. D.; Sinhoreti, M.A. C.; Braz, R.; Montes, M., Effect of dentin pre-treatment with a caseinphosphopeptide-amorphous calcium phosphate (CPP-ACP) paste on dentin bondstrength in tridimensional cavities[J]. Acta Odontologica Scandinavica,2013,71,271-277.
[37] Cheng, G.; Wang, Z. G.; Liu, Y. L.; Zhang, J. L.; Sun, D. H.; Ni, J. Z.,Phosphoprotein/Phosphopeptide Enrichment and Analysis Based on NanostructuredMaterials[J]. Progress in Chemistry,2013,25,620-632.
[38] Li, J. T.; Zhang, J. L.; He, H.; Ma, Z. L.; Nie, Z. K.; Wang, Z. Z.; Xu, X. G.,Apoptosis in human hepatoma HepG2cells induced by corn peptides and its anti-tumorefficacy in H22tumor bearing mice[J]. Food and Chemical Toxicology,2013,51,297-305.
[39] Ma, Z. L.; Zhang, W. J.; Yu, G. C.; He, H.; Zhang, Y., The primary structureidentification of a corn peptide facilitating alcohol metabolism by HPLC-MS/MS[J].Peptides,2012,37,138-143.
[40] Roblet, C.; Doyen, A.; Amiot, J.; Bazinet, L., Impact of pH on ultrafiltrationmembrane selectivity during electrodialysis with ultrafiltration membrane (EDUF)purification of soy peptides from a complex matrix[J]. Journal of Membrane Science,2013,435,207-217.
[41] Singh, A.; Banerjee, R., Peptide enriched functional food adjunct from soy whey: Astatistical optimization study[J]. Food Science and Biotechnology,2013,22,65-71.
[42] Tsou, M. J.; Kao, F. J.; Lu, H. C.; Kao, H. C.; Chiang, W. D., Purification andidentification of lipolysis-stimulating peptides derived from enzymatic hydrolysis of soyprotein[J]. Food Chemistry,2013,138,1454-1460.
[43] Nogata, Y.; Saito, T.; Abe, D.; Yoshida, A.; Ohta, H., Storage Stability of WheatBran Proteases for Producing Angiotensin I-converting Enzyme Inhibitory Peptides[J].Food Science and Technology Research,2013,19,313-317.
[44] Prandi, B.; Faccini, A.; Tedeschi, T.; Galaverna, G.; Sforza, S., LC/MS analysis ofproteolytic peptides in wheat extracts for determining the content of the allergenamylase/trypsin inhibitor CM3: Influence of growing area and variety[J]. FoodChemistry,2013,140,141-146.
[45] Campos-Quevedo, N.; Rosales-Mendoza, S.; Paz-Maldonado, L. M. T.;Martinez-Salgado, L.; Guevara-Arauza, J. C.; Soria-Guerra, R. E., Production ofmilk-derived bioactive peptides as precursor chimeric proteins in chloroplasts ofChlamydomonas reinhardtii[J]. Plant Cell Tissue and Organ Culture,2013,113,217-225.
[46] Fleminger, G.; Ragones, H.; Merin, U.; Silanikove, N.; Leitner, G., Low molecularmass peptides generated by hydrolysis of casein impair rennet coagulation of milk[J].International Dairy Journal,2013,30,74-78.
[47] Picariello, G.; Iacomino, G.; Mamone, G.; Ferranti, P.; Fierro, O.; Gianfrani, C.; DiLuccia, A.; Addeo, F., Transport across Caco-2monolayers of peptides arising from invitro digestion of bovine milk proteins[J]. Food Chemistry,2013,139,203-212.
[48] Pan, D. D.; Lu, H. Q.; Zeng, X. Q., A Newly Isolated Ca Binding Peptide fromWhey Protein[J]. International Journal of Food Properties,2013,16,1127-1134.
[49] Saiga, A.; Okumura, T.; Makihara, T.; Katsuta, S.; Shimizu, T.; Yamada, R.;Nishimura, T., Angiotensin I-converting enzyme inhibitory peptides in a hydrolyzedchicken breast muscle extract[J]. Journal of Agricultural and Food Chemistry,2003,51,1741-1745.
[50] Gomez-Ruiz, J. A.; Ramos, M.; Recio, I., Angiotensin-convertingenzyme-inhibitory peptides in Manchego cheeses manufactured with different startercultures[J]. International Dairy Journal,2002,12,697-706.
[51] Gimenez, B.; Aleman, A.; Montero, P.; Gomez-Guillen, M. C., Antioxidant andfunctional properties of gelatin hydrolysates obtained from skin of sole and squid[J].Food Chemistry,2009,114,976-983.
[52] Wang, S.; Agyare, K.; Damodaran, S., Optimisation of hydrolysis conditions andfractionation of peptide cryoprotectants from gelatin hydrolysate[J]. FoodChemistry,2009,115,620-630.
[53] Ranathunga, S.; Rajapakse, N.; Kim, S. K., Purification and characterization ofantioxidative peptide derived from muscle of conger eel (Conger myriaster)[J].European Food Research and Technology,2006,222,310-315.
[54] Sheih, I. C.; Fang, T. J.; Wu, T. K., Isolation and characterisation of a novelangiotensin I-converting enzyme (ACE) inhibitory peptide from the algae proteinwaste[J]. Food Chemistry,2009,115,279-284.
[55] Jung, W. K.; Jo, H. Y.; Qian, Z. J.; Jeong, Y. J.; Park, S. G.; Choi, I. W.; A novelanticoagulant protein with high affinity to blood coagulation factor Va from Tegillarcagranosa[J]. Journal of Biochemistry and Molecular Biology,2007,40,832-838.
[56] Sheih, I. C.; Wu, T. K.; Fang, T. J., Antioxidant properties of a new antioxidativepeptide from algae protein waste hydrolysate in different oxidation systems[J].Bioresource Technology,2009,100,3419-3425.
[57] Lee, J. K.; Hong, S.; Jeon, J. K.; Kim, S. K.; Byun, H. G., Purification andcharacterization of angiotensin I converting enzyme inhibitory peptides from the rotifer,Brachionus rotundiformis[J]. Bioresource Technology2009,100,5255-5259.
[58] Kim, S. Y.; Je, J. Y.; Kim, S. K., Purification and characterization of antioxidantpeptide from hoki (Johnius belengerii) frame protein by gastrointestinal digestion[J].Journal of Nutritional Biochemistry,2007,18,31-38.
[59] Lee, S. H.; Qian, Z. J.; Kim, S. K., A novel angiotensin I converting enzymeinhibitory peptide from tuna frame protein hydrolysate and its antihypertensive effect inspontaneously hypertensive rats[J]. Food Chemistry,2010,118,96-102.
[60] Jun, S. Y.; Park, P. J.; Jung, W. K.; Kim, S. K., Purification and characterization ofan antioxidative peptide from enzymatic hydrolysate of yellowfin sole (Limanda aspera)frame protein[J]. European Food Research and Technology,2004,219,20-26.
[61] Jung, W. K.; Karawita, R.; Heo, S. J.; Lee, B. J.; Kim, S. K.; Jeon, Y. J., Recoveryof a novel Ca-binding peptide from Alaska Pollack (Theragra chalcogramma) backboneby pepsinolytic hydrolysis[J]. Process Biochemistry,2006,41,2097-2100.
[62] Wilson, J.; Hayes, M.; Carney, B., Angiotensin-I-converting enzyme and prolylendopeptidase inhibitory peptides from natural sources with a focus on marineprocessing by-products[J]. Food Chemistry,2011,129,235-244.
[63] Lin, K. H.; Jaw, C. G.; Yen, J. H.; Wang, Y. S., Molecular connectivity indices forpredicting bioactivities of substituted nitrobenzene and aniline compounds[J].Ecotoxicology and Environmental Safety,2009,72,1942-1949.
[64] Liu, Y.; Ke, Z.; Cui, J.; Chen, W.-H.; Ma, L.; Wang, B., Synthesis, inhibitoryactivities, and QSAR study of xanthone derivatives as alpha-glucosidase inhibitors[J].Bioorganic&Medicinal Chemistry,2008,16,7185-7192.
[65] Majumder, K.; Wu, J. P., A new approach for identification of novelantihypertensive peptides from egg proteins by QSAR and bioinformatics[J]. FoodResearch International,2010,43,1371-1378.
[66] Pripp, A. H., Docking and virtual screening of ACE inhibitory dipeptides[J].European Food Research and Technology,2007,225,589-592.
[67] Pripp, A. H.; Isaksson, T.; Stepaniak, L.; Sorhaug, T., Quantitative structure-activityrelationship modelling of ACE-inhibitory peptides derived from milk proteins[J].European Food Research and Technology2004,219,579-583.
[68] Pripp, A. H.; Isaksson, T.; Stepaniak, L.; Sorhaug, T.; Ardo, Y., Quantitativestructure activity relationship modelling of peptides and proteins as a tool in foodscience[J]. Trends in Food Science&Technology,2005,16,484-494.
[69] Maryam, P.; Davood, A.; Niusha, S., Tertiary structure prediction ofalpha-Glucosidase and inhibition properties of N-(phenoxydecyl)phthalimidederivatives by modeling, Docking and QSAR[J]. Clinical Biochemistry2011,44,S101-S102.
[70] Wu, J. P.; Aluko, R. E.; Nakai, S., Structural requirements of angiotensinI-converting enzyme inhibitory peptides: Quantitative structure-activity relationshipmodeling of peptides containing4-10amino acid residues[J]. Qsar&CombinatorialScience,2006,25,873-880.
[71] Choi, I.; Jung, C.; Choi, H.; Kim, C.; Ha, H., Effectiveness of phosvitin peptides onenhancing bioavailability of calcium and its accumulation in bones[J]. Food Chemistry,2005,93,577-583.
[72] Mine, Y., Egg proteins and peptides in human health-chemistry, bioactivity andproduction[J]. Current Pharmaceutical Design,2007,13,875-884.
[73] Xu, C. N.; Yang, C. B.; Yin, Y. G.; Liu, J. B.; Mine, Y., Phosphopeptides (PPPs)from hen egg yolk phosvitin exert anti-inflammatory activity via modulation of cytokineexpression[J]. Journal of Functional Foods,2012,4,718-726.
[74] Fujita, H.; Sasaki, R.; Yoshikawa, M., Potentiation of the antihypertensive activityof orally administered ovokinin, a vasorelaxing peptide derived from ovalbumin, byemulsification in egg phosphatidylcholine[J]. Bioscience Biotechnology andBiochemistry,1995,59,2344-2345.
[75] You, S. J.; Udenigwe, C. C.; Aluko, R. E.; Wu, J. P., Multifunctional peptides fromegg white lysozyme[J]. Food Research International,2010,43,848-855.
[76] Davalos, A.; Miguel, M.; Bartolome, B.; Lopez-Fandino, R., Antioxidant activity ofpeptides derived from egg white proteins by enzymatic hydrolysis[J]. Journal of FoodProtection,2004,67,1939-1944.
[77] Wang, J.; Liang, Y.; Omana, D. A.; Kav, N. N. V.; Wu, J., Proteomics Analysis ofEgg White Proteins from Different Egg Varieties[J]. Journal of Agricultural and FoodChemistry,2012,60,272-282.
[78] Wu, J. P.; Aluko, R. E.; Muir, A. D., Improved method for direct high-performanceliquid chromatography assay of angiotensin-converting enzyme-catalyzed reactions[J].Journal of Chromatography A,2002,950,125-130.
[79] Wu, J. P.; Aluko, R. E.; Muir, A. D., Purification of angiotensin I-convertingenzyme-inhibitory peptides from the enzymatic hydrolysate of defatted canola meal[J].Food Chemistry,2008,111,942-950.
[80] Wu, J.; Acero-Lopez, A., Ovotransferrin: Structure, bioactivities, and preparation[J].Food Research International,2012,46,480-487.
[81] Wu, J. P.; Aluko, R. E., Quantitative structure-activity relationship study of bitterdi-and tri-peptides including relationship with angiotensin I-converting enzymeinhibitory activity[J]. Journal of Peptide Science,2007,13,63-69.
[82] Wu, J. P.; Aluko, R. E.; Nakai, S., Structural requirements of angiotensinI-converting enzyme inhibitory peptides: Quantitative structure-activity relationshipstudy of di-and tripeptides[J]. Journal of Agricultural and Food Chemistry,2006,54,732-738.
[83] Lv, L.; Chi, Y. J., Improvement of functional properties of ovalbuminphosphorylated by dry-heating in the presence of pyrophosphate[J]. European FoodResearch and Technology,2012,235,981-987.
[84] Chen, C.; Chi, Y. J., Antioxidant, ace inhibitory activities and functional propertiesof egg white protein hydrolysate[J]. Journal of Food Biochemistry,2012,36,383-394.
[85] Chen, C.; Chi, Y. J.; Lv, L., Purification and identification of antioxidant peptidesfrom egg white protein hydrolysate[J]. Amino Acids,2012,43,457-466.
[86] Liu, J. B.; Yu, Z. P.; Zhao, W. Z.; Lin, S. Y.; Wang, E. L.; Zhang, Y.; Hao, H.; Chen,F., Isolation and identification of angiotensin-converting enzyme inhibitory peptidesfrom egg white protein hydrolysates[J]. Food Chemistry,2010,122,1159-1163.
[87] Yu, Z. P.; Zhao, W. Z.; Liu, J. B.; Lu, J.; Chen, F., QIGLF, a novel angiotensinI-converting enzyme-inhibitory peptide from egg white protein[J]. Journal of theScience of Food and Agriculture,2011,91,921-926.
[88] Yu, Z. P.; Liu, B. Q.; Zhao, W. Z.; Yin, Y. G.; Liu, J. B.; Chen, F., Primary andsecondary structure of novel ACE-inhibitory peptides from egg white protein[J]. FoodChemistry2012,133,315-322.
[89] Liu Lijun,Zhu Chuanhe,Zhao Zheng. Analyzing molecular weight distribution ofwhey protein hydrolysates[J]. Food and bioproducts processing,2008,86(C1):1-6.
[90]刘静波,于志鹏,赵文竹,等.蛋清源ACE抑制肽结构鉴定及其稳定性[J].吉林大学学报(工学版),2011,(02):579-584.
[91]于志鹏.蛋清源ACE抑制肽的结构鉴定及稳定性研究[D].长春:吉林大学,2011.
[92] Yu, Z.; Yin, Y.; Zhao, W.; Wang, F.; Yu, Y.; Liu, B.; Liu, J.; Chen, F.,Characterization of ACE-Inhibitory Peptide Associated with Antioxidant andAnticoagulation Properties[J]. Journal of Food Science,2011,76, C1149-C1155.
[93]张超,张晖,赵晓燕,马越,姚惠源.使用圆二色性光谱和红外光谱研究冬小麦麸皮抗冻蛋白的二级结构[J].光谱学与光谱分析,2009,29(7):1764-1767.
[94] Murariu, M.; Dragan, E. S.; Drochioiu, G., IR, MS and CD Investigations onSeveral Conformationally-Different Histidine Peptides[J]. International Journal ofPeptide Research and Therapeutics,2009,15,303-311.
[95] Carbonaro, M.; Nucara, A., Secondary structure of food proteins by Fouriertransform spectroscopy in the mid-infrared region[J]. Amino Acids,2010,38,679-690.
[96] Miguel, M.; Recio, I.; Gomez-Ruiz, J. A.; Ramos, M.; Lopez-Fandino, R.,Angiotensin I-converting enzyme inhibitory activity of peptides derived from egg whiteproteins by enzymatic hydrolysis[J]. Journal of Food Protection,2004,67,1914-1920.
[97] Marcuse, R., Antioxidative effect of amino-acids[J]. Nature,1960,186,886-887.
[98] Chen, C.; Chi, Y. J., antioxidant, ace inhibitory activities and functional propertiesof egg white protein hydrolysate[J].Journal of Food Biochemistry,2012,36(4),383-394.
[99]Chen, C.; Chi, Y. J.; Zhao, M. Y.; Lv, L., Purification and identification ofantioxidant peptides from egg white protein hydrolysate[J]. Amino Acids,2012,43(1),457-466.
[100] You, S. J.; Udenigwe, C. C.; Aluko, R. E.; Wu, J. P., Multifunctional peptides fromegg white lysozyme[J]. Food Research International,2010,43(3),848-855.
[101] Li, B.; Chen, F.; Wang, X.; Ji, B. P.; Wu, Y. N., Isolation and identification ofantioxidative peptides from porcine collagen hydrolysate by consecutivechromatography and electrospray ionization-mass spectrometry[J]. Food Chemistry,2007,102(4),1135-1143.
[102] Zhong, S.; Ma, C.; Lin, Y. C.; Luo, Y., Antioxidant properties of peptide fractionsfrom silver carp (Hypophthalmichthys molitrix) processing by-product proteinhydrolysates evaluated by electron spin resonance spectrometry[J]. Food Chemistry,2011,126(4),1636-1642.
[103]王嘉荣，滕达，田子罡，等.功能性抗氧化肽制备与机制研究进展[J].天然产物研究与开发,2008,20:371-375.
[104] Kokubu, T.; Ueda, E.; Fujimoto, S.; Hiwada, K.; Kato, A.; Akutsu, H.; Yamamura,Y.; Saito, S.; Mizoguch.T, Peptide inhibitors of renin-angiotensin system[J]. Nature,1968,217(5127),456-&.
[105] Ishiguro, K.; Sameshima, Y.; Kume, T.; Ikeda, K.-i.; Matsumoto, J.; Yoshimoto,M., Hypotensive effect of a sweetpotato protein digest in spontaneously hypertensiverats and purification of angiotensin I-converting enzyme inhibitory peptides[J]. FoodChemistry,2012,131(3),774-779.
[106] Guang, C.; Phillips, R. D.; Jiang, B.; Milani, F., Three key proteases-angiotensin-l-converting enzyme (ACE), ACE2and renin-within and beyond therenin-angiotensin system[J]. Archives of Cardiovascular Diseases,2012,105(6-7),373-385.
[107] Lin, L.; Lv, S.; Li, B., Angiotensin-I-converting enzyme (ACE)-inhibitory andantihypertensive properties of squid skin gelatin hydrolysates[J]. Food Chemistry,2012,131(1),225-230.
[108] Li, Y.; Zhou, J.; Huang, K.; Sun, Y.; Zeng, X., Purification of a Novel AngiotensinI-Converting Enzyme (ACE) Inhibitory Peptide with an Antihypertensive Effect fromLoach (Misgurnus anguillicaudatus)[J]. Journal of agricultural and food chemistry,2012,60(5),1320-5.
[109] Lu, J.; Sawano, Y.; Miyakawa, T.; Xue, Y.-L.; Cai, M.-Y.; Egashira, Y.; Ren, D.-F.;Tanokura, M., One-Week Antihypertensive Effect of Ile-Gln-Pro in SpontaneouslyHypertensive Rats[J]. Journal of Agricultural and Food Chemistry,2011,59(2),559-563.
[110] Wang, X.; Wang, L.; Cheng, X.; Zhou, J.; Tang, X.; Mao, X. Y.,Hypertension-attenuating effect of whey protein hydrolysate on spontaneouslyhypertensive rats[J]. Food Chemistry,2012,134(1),122-126.
[111] Li, Y.; Zhou, J.; Huang, K.; Sun, Y.; Zeng, X., Purification of a Novel AngiotensinI-Converting Enzyme (ACE) Inhibitory Peptide with an Antihypertensive Effect fromLoach (Misgurnus anguillicaudatus)[J]. Journal of Agricultural and Food Chemistry,2012,60(5),1320-1325.
[112] Ghosh, S.; Ahire, M.; Patil, S.; Jabgunde, A.; Bhat Dusane, M.; Joshi, B. N.;Pardesi, K.; Jachak, S.; Dhavale, D. D.; Chopade, B. A., Antidiabetic Activity of Gnidiaglauca and Dioscorea bulbifera: Potent Amylase and Glucosidase Inhibitors[J].Evidence-based complementary and alternative medicine: eCAM,2012,2012,929051.
[113] Lee, H. J.; Lee, H.-S.; Choi, J. W.; Ra, K. S.; Kim, J.-M.; Suh, H. J., NovelTripeptides with alpha-Glucosidase Inhibitory Activity Isolated from Silk CocoonHydrolysate[J]. Journal of agricultural and food chemistry,2011,59(21),11522-5.
[114] Kim, Y. M.; Wang, M. H.; Rhee, H. I., A novel alpha-glucosidase inhibitor frompine bark[J]. Carbohydrate Research,2004,339(3),715-717.
[115] Yu, Z. P.; Yin, Y. G.; Zhao, W. Z.; Yu, Y. D.; Liu, B. Q.; Liu, J. B.; Chen, F., Novelpeptides derived from egg white protein inhibiting alpha-glucosidase[J]. FoodChemistry,2011,129(4),1376-1382.
[116]杨万根.蛋清蛋白水解物的制备结构及其生物活性的研究[D].无锡，江南大学，2008.
[117] Yu, Z. P.; Yin, Y. G.; Zhao, W. Z.; Liu, J. B.; Chen, F., Anti-diabetic activitypeptides from albumin against alpha-glucosidase and alpha-amylase[J]. Food Chemistry,2012,135(3),2078-2085.
[118]王本祥，杨明，金玉莲，崔志勇，王岩.人参多肽降血糖作用[J].药学学报，1990，25(6):401-405.
[119]吴鸿,邹大进.糖尿病新药应用进展[J].中国实用内科杂志.2009,29(10):962-963.
[120]张瑞东,迟玉杰,陈晨,阮长青.蛋清蛋白酶解物抗氧化与血管紧张素转换酶抑制活性的研究[J].营养学报,2010,32(4).
[121]李亚云,赵新淮.两步法制备高活性酪蛋白ACE抑制肽的工艺参数研究[J].食品科学，31(10):6-11.
[122] Asoodeh, A.; Yazdi, M. M.; Chamani, J., Purification and characterisation ofangiotensin I converting enzyme inhibitory peptides from lysozyme hydrolysates[J].Food Chemistry,2012,131(1),291-295.
[123] Chen, C.; Chi, Y. J., Antioxidant, ace inhibitory activities and functional propertiesof egg white protein hydrolysate[J]. Journal of Food Biochemistry,2012,36(4),383-394.
[124] Garcia-Redondo, A. B.; Roque, F. R.; Miguel, M.; Lopez-Fandino, R.; Salaices,M., Vascular effects of egg white-derived peptides in resistance arteries from rats.Structure-activity relationships[J]. Journal of the Science of Food and Agriculture,2010,90(12),1988-1993.
[125] Miguel, M.; Recio, I.; Gomez-Ruiz, J. A.; Ramos, M.; Lopez-Fandino, R.,Angiotensin I-converting enzyme inhibitory activity of peptides derived from egg whiteproteins by enzymatic hydrolysis[J]. Journal of Food Protection,2004,67(9),1914-1920.
[126] Miguel, M.; Lopez-Fandino, R.; Ramos, M.; Aleixandre, A., Short-term effect ofegg-white hydrolysate products on the arterial blood pressure of hypertensive rats[J].British Journal of Nutrition,2005,94(5),731-737.
[127] Miguel, M.; Lopez-Fandino, R.; Ramos, M.; Aleixandre, A., Long-term intake ofegg white hydrolysate attenuates the development of hypertension in spontaneouslyhypertensive rats[J]. Life Sciences,2006,78(25),2960-2966.
[128]蒋菁莉,任发政.食源性降血压肽的评价方法[J].中国乳品工业,2006，34(6):36-39.
[129]卢静.大豆肽对大鼠的心血管效应及机制研究[D].长春：吉林大学，2006.
[130] Wang, X.; Wang, L.; Cheng, X.; Zhou, J.; Tang, X.; Mao, X. Y.,Hypertension-attenuating effect of whey protein hydrolysate on spontaneouslyhypertensive rats[J]. Food Chemistry2012,134(1),122-126.
[131]鲁军.螺旋藻活性肽的纯化鉴定及降压护肝机制研究[D].北京：北京林业大学，2009.
[132] He, R.; Alashi, A.; Malomo, S. A.; Girgih, A. T.; Chao, D. F.; Ju, X. R.; Aluko, R.E., Antihypertensive and free radical scavenging properties of enzymatic rapeseedprotein hydrolysates[J]. Food Chemistry2013,141(1),153-159.
[133] Fujita, H.; Yokoyama, K.; Yoshikawa, M., Classification and antihypertensiveactivity of angiotensin I-converting enzyme inhibitory peptides derived from foodproteins[J]. Journal of Food Science2000,65(4),564-569.
[134] Fujita, H.; Sasaki, R.; Yoshikawa, M., Potentiation of the antihypertensive activityof orally administered ovokinin, a vasorelaxing peptide derived from ovalbumin, byemulsification in egg phosphatidylcholine[J]. Bioscience Biotechnology andBiochemistry1995,59(12),2344-2345.
[135] Miguel, M.; Alvarez, Y.; Lopez-Fandino, R.; Alonso, M. J.; Salaices, M.,Vasodilator effects of peptides derived from egg white proteins[J]. Regulatory Peptides2007,140(3),131-135.
[136] Miguel, M.; Lopez-Fandino, R.; Aleixandre, M. A., Effect of the hydrolysateobtained from egg white with pepsin on the development of hypertension ofspontaneously hypertensive rats[J]. Ciencia y Tecnologia Alimentaria2005,4(5),368-372.
[137]陈季旺,刘珊珊,蔡广霞,吴永宁.食物源血管紧张素转化酶抑制三肽的定量构效关系[J].食品科学,2013,34（9）：19-23.
[138] Kearney, P. M.; Whelton, M.; Reynolds, K.; Muntner, P.; Whelton, P. K.; He, J.,Global burden of hypertension: analysis of worldwide data[J]. Lancet2005,365(9455),217-223.
[139] Violle, N.; Messaoudi, M.; Lefranc-Millot, C.; Desor, D.; Nejdi, A.; Demagny, B.;Schroeder, H., Ethological comparison of the effects of a bovine alpha(s1)-casein tryptichydrolysate and diazepam on the behaviour of rats in two models of anxiety[J].Pharmacology Biochemistry and Behavior2006,84(3),517-523;
[140] Walf, A. A.; Frye, C. A., The use of the elevated plus maze as an assay ofanxiety-related behavior in rodents[J]. Nature Protocols2007,2(2),322-328.
[141]刘爽,夏杨毅,尚永彪.鸡蛋蛋清蛋白酶解产物的研究进展[J].食品科学,2013,34(15):409-414.
[142] Hooge, R.; De Deyn, P. P., Applications of the Morris water maze in the study oflearning and memory[J]. Brain Research Reviews2001,36(1),60-90.
[143] McNamara, R. K.; Skelton, R. W., The neuropharmacological and neurochemicalbasis of place learning in the morris water maze[J]. Brain Research Reviews1993,18(1),33-49.
[144] Capozziello, S.; De Laurentis, M.; Nojiri, S.; Odintsov, S. D., Classifying andavoiding singularities in the alternative gravity dark energy models[J]. Physical ReviewD2009,79(12).
[145] Gilbert, P.; McEwan, K.; Bellew, R.; Mills, A.; Gale, C., The dark side ofcompetition: How competitive behaviour and striving to avoid inferiority are linked todepression, anxiety, stress and self-harm[J]. Psychology and Psychotherapy-TheoryResearch and Practice2009,82(2),123-136.
[146] Schijven, P.; Mulken, O., Avoiding dark states in open quantum systems bytailored initializations[J]. Physical Review E2012,85(6).
[147]许崇涛,李慧,林凌云.快眼动睡眠剥夺对抑郁模型大鼠自主活动及海马5-羟色胺含量的影响[J].中国行为医学科学,2004,13(4):381-383.
[148]刘屏,陈世铭,侯津详.高架十字迷宫和Vogel饮水冲突法用于吗啡依赖动物焦虑评价[J].中国药理学报,1997,13(6):563-565.
[149]宗绍波,魏盛,苏云祥,乔明琦.焦虑大鼠模型高架十字迷宫实验的复测信度检验及参数相关性分析[J].中国医药导报,2011,8(30):5-7.
[150]陈罗西,郭玲玲,李亮. Morris圆形水迷宫的应用及相关检测指标分析[J].辽宁中医药大学学报,2008,10(8):55-57.
[151]胡镜清,温泽淮,赖世隆. Morris水迷宫检测的记忆属性与方法学初探[J].广州中医药大学学报,2000,17(2):117-119.