海蜇生殖腺酶解肽的抗氧化活性和ACE抑制活性研究

为实现海蜇加工副产物生殖腺的高值化利用,以海蜇性腺脱脂粉为原料,选用中性蛋白酶、木瓜蛋白酶、胰蛋白酶和碱性蛋白酶分别对海蜇生殖腺进行水解,以酶解产物（JGHP）的水解度、DPPH清除活性和ACE抑制活性为指标,选择制备海蜇生殖腺活性肽的工具酶。经超滤膜将酶解产物分成JGPH-P1（大于3000 u）、P2（1000³000 u）、P3（小于1000 u）三个部分,分别测定三部分的DPPH清除活性和ACE抑制活性。结果表明,中性蛋白酶水解所得产物的水解度、DPPH清除率和ACE抑制活性均优于其他三种蛋白酶;JGHP超滤后,其中组分JGHP-P3的活性较高。利用电子自旋共振（ESR）技术检测JGHP-P3对DPPH·、·OH、O-2·的清除活性。其ACE抑制活性IC50为1.06 mg/m L,DPPH·、·OH、O-2·的清除活性IC50分别为0.38、0.63、0.59 mg/m L。由此可见,经中性蛋白酶水解得到的JGPH中小于1000 u的组分具有较高的抗氧化和ACE抑制活性。      

Purification and characterization of angiotensin I converting enzyme inhibitory peptides from jellyfish Rhopilema esculentum

Two angiotensin I converting enzyme (ACE) inhibitory peptides were isolated from jellyfish Rhopilema esculentum protein hydrolysates prepared with pepsin and papain. Consecutive purification methods, including ion-exchange chromatography, size-exclusion chromatography and reverse-phase high-performance liquid chromatography, were used for isolation of ACE inhibitory peptides. The amino acid sequence of the peptides was identified as Gln-Pro-Gly-Pro-Thr and Gly-Asp-Ile-Gly-Tyr, respectively, and the IC₅₀ value of the purified peptides for ACE inhibitory activity was 80.67 μM and 32.56 μM. The purified peptides were evaluated for the antihypertensive effect in spontaneously hypertensive rats (SHR) after oral administration. Blood pressure decreased significantly after ingestion of the peptides. The results suggest that peptides derived from jellyfish R. esculentum may be beneficial as antihypertensive compounds in functional food resources.     

海蜇胶原蛋白提取工艺及促愈合作用的研究

通过响应面法优化海蜇胶原蛋白的提取工艺,并通过小鼠皮肤创伤模型研究海蜇胶原蛋白的促愈合作用。通过单因素试验选取因素与水平,根据Box-Benhnken 中心组合试验设计原理,在单因素基础上采用三因素三水平响应面分析法(RSM),对海蜇胶原蛋白的酸法提取工艺进行优化;采用小鼠皮肤创伤模型,创面外用胶原蛋白,并以云南白药处理为阳性对照,通过十字测量法测量创面大小并计算创面愈合率。结果显示海蜇胶原蛋白的最佳提取工艺条件为提取时间62.64h、液料比23.6:1,醋酸浓度0.562mol/L;动物实验表明海蜇胶原蛋白能够明显缩短伤口愈合时间,对创面修复有促进作用。响应面法优化得到的提取条件准确可靠;海蜇胶原蛋白具有明显的促愈合作用。     

海蜇伞部酶促溶性胶原蛋白的热变性动力学

为阐明海蜇伞部酶促溶性胶原蛋白(pepsin-solubilized collagen,PSC)的热变性反应机理,以保持完整三螺旋结构的PSC为研究对象,通过微量热仪测定不同升温速率条件下PSC的变性温度,以及采用34、35、36、37、38、39℃加热不同时间后的PSC残存率,并进行热变性动力学分析。结果表明,海蜇伞部PSC对热变化敏感,随着加热温度升高,单位时间内提高的热量增加,使海蜇伞部PSC变性速率加快,完成变性时间缩短;随着升温速率的减慢,吸热峰逐渐向低温区移动,即变性温度随升温速率的减慢而降低,但升温速率的变化对反应热并无显著影响。反应级数为0.9的回归方程能够较好地描述PSC热变性过程,在恒温34、35、36、37、38℃及39℃的条件下,PSC变性的D90值(90%蛋白变性所需时间)分别为53.76、26.11、15.75、4.89、4.26、2.55 min,Z90值(D值降低90%的温度变化)为3.69℃,表观活化能为481.90 k J/mol。研究结果可为海蜇胶原蛋白的进一步开发利用提供理论参考。     

Effects of Collagen and Collagen Hydrolysate from Jellyfish (Rhopilema esculentum) on Mice Skin Photoaging Induced by UV Irradiation

ABSTRACT:  Collagen (JC) was extracted from jellyfish ( Rhopilema esculentum ) and hydrolyzed to prepare collagen hydrolysate (JCH). The protective effects of JC and JCH against UV-induced damages to mice skin were evaluated and compared in this article. JC and JCH could alleviate the UV-induced abnormal changes of antioxidative indicators, including the superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) activities and the contents of glutathione (GSH) and malondiaidehyde (MDA). JC and JCH could protect skin lipid and collagen from the UV radiation damages. Furthermore, the changes of total ceramide and glycosaminoglycan in skin were recovered significantly by JC and JCH. The action mechanisms mainly involved the antioxidative properties and the repairing to endogenous collagen synthesis of JC and JCH  in vivo . JCH with the lower molecular weight showed much higher effects than JC. The results indicated that JCH was a novel antiphotoaging agent from natural resources.     

Isolation and characterisation of collagen from the ribbon jellyfish (Chrysaora sp.)

Pepsin‐solubilised collagen from the ribbon jellyfish (Chrysaora sp., morphotype 1) umbrella (JPSC) was isolated and characterised. The yield of collagen varied (9–19%, based on ash‐free dry weight) depending on the amount of pepsin used. Type II collagen was the major component of extracted collagen. The peptide map of JPSC differed from that of standard collagen type II, which indicates their different primary structures. FTIR spectra of JPSC, however, did not differ significantly from those of type II collagen. The T_max of JPSC was 37.38 °C, which is higher than that of other marine collagens. Glycine was the main amino acid in JPSC (320 residues per 1000 residues), followed by glutamic acid, alanine, proline, aspartic acid and hydroxyproline. The isoelectric point of JPSC was 6.64. These results indicate that this jellyfish species has the potential to be a marine source of type II collagen that can be used in place of land‐based sources.     

Antioxidant and tyrosinase inhibitory activities of a novel chitosan–phloroglucinol conjugate

A novel chitosan–phloroglucinol conjugate was developed by conjugating phloroglucinol onto a chitosan backbone. The chitosan–phloroglucinol conjugate was characterised by ¹H nuclear magnetic resonance (NMR), and the NMR spectra confirmed the conjugation. Antioxidant and tyrosinase inhibitory activities of the chitosan–phloroglucinol conjugate were investigated. The chitosan–phloroglucinol conjugate showed strong 2,2‐diphenyl‐1‐picrylhydrazyl (DPPH), hydrogen peroxide and ABTS⁺ radical scavenging activities as well as reducing power compared with those of the unmodified chitosan (P < 0.05). The formation of malondiadehyde (MDA) as an indicator of lipid peroxidation in a linoleic acid emulsion was 30.56 μM in the absence of the chitosan–phloroglucinol conjugate after 4 days incubation, whereas MDA was 4.14 μM in the presence of the chitosan–phloroglucinol conjugate (P < 0.05). The activity was higher than that of ascorbic acid, which is currently used as a food preservative. Moreover, the chitosan–phloroglucinol conjugate inhibited 56.30% tyrosinase activity, which is responsible for browning of foods, and acted as non‐competitive inhibitor. Taken together, the chitosan–phloroglucinol conjugate may have potential for application in functional foods and/or as a food preservative.     

不同方法提取沙海蜇皮胶原蛋白的对比分析

以沙海蜇皮为实验原料,分别用胃蛋白酶提取法、热水提取法和酸提取法进行了胶原蛋白的提取,得到酶溶性胶原蛋白(PSC)、热水抽提胶原蛋白(HEG)和酸溶性胶原蛋白(ASC),并分析了三种胶原蛋白的理化性质及功能特性。结果表明:PSC、HEG和ASC中羟脯氨酸含量分别为5.77%,5.53%和9.20%;平均分子量分别为34、52、38ku。紫外图谱扫描和红外扫描光谱表明,三种方法提取的胶原蛋白结构相似,但略有差别。PSC、HEG、ASC乳化指数依次为:5.07、25.90、3.26m2/g;稳定性依次为:24.68、21.89、88.5min。吸湿性和保湿性最好的样品均为PSC。     

海蜇胶原蛋白肽的生物活性研究

以海蜇加工下脚料为原料提取胶原蛋白,酶解制备胶原蛋白肽并对酶解液进行抽滤分离,并对所得不同分子量的胶原蛋白肽进行生物活性研究。结果表明:抽滤分离到四种不同分子量的胶原蛋白肽:分子量>10 kDa(JCP1)、分子量3~10 kDa(JCP2)、分子量1~3 kDa(JCP3)和<1 kDa(JCP4)。生物活性研究表明,JCP1具有最强的超氧阴离子自由基清除能力和酪氨酸酶双酚酶抑制效果,其IC₅₀值分别为22.6和11.96 mg/mL,JCP1对酪氨酸酶的抑制类型为可逆混合竞争型抑制;JCP3具有最强的还原力、羟自由基、2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS)自由基清除能力和血管紧张素转换酶(ACE)抑制活性,其IC₅₀值分别为14.9、2.21、0.61和15.8 mg/mL;JCP4具有最强的1,1-二苯基-2-三硝基苯肼(DPPH)自由基清除能力,其IC₅₀值为28.3 mg/mL。     

海蜇不同组织营养组成分析及评价

为了实现海蜇资源的高值化综合加工利用,对新鲜海蜇不同组织(伞体、胃柱、肩板、口腕、棒状附属器、生殖腺和环肌)的基本营养组成、氨基酸组成、脂质组成及脂肪酸组成进行分析与评价。结果表明,海蜇不同组织水分均在93%以上,其中胃柱水分含量最高(97.93%);灰分含量也较高,其中伞体、棒状附属器、胃柱和口腕的灰分含量均高于50%(干质量)。以除去灰分后的干质量计,棒状附属器和环肌的粗脂肪含量较高,分别为18.80%和18.76%,其他组织的粗脂肪含量均较低;所有组织的粗蛋白含量均较高,特别是伞体、肩板和口腕,粗蛋白含量达62.81%~80.94%;总糖含量均较低,为6.28%~13.36%。海蜇的生殖腺和环肌组织检出20种氨基酸,必需氨基酸含量高于其他组织,且不同组织的必需氨基酸均在25%以上。海蜇各组织的磷脂含量较多,胆固醇含量相对较少;伞体和棒状附属器的饱和脂肪酸含量多于不饱和脂肪酸,其他组织相反。以上结果说明,海蜇各组织均含有较为丰富的营养价值,且各个组织的营养价值具有一定的特点,可针对不同组织的营养特点进一步开发与利用。     

大豆肽、小麦肽、胶原肽在运动营养领域的研究进展（网络首发、推荐阅读）

综述运动营养市场发展趋势和前景,以及肽在运动营养领域中的研究进展及功能作用,主要包括提供运动所需蛋白质和必需氨基酸（EAA）、缓解疲劳、促进肌肉损伤修复、促进脂肪代谢、提升运动表现能力等。重点阐述大豆肽、小麦肽和胶原肽的分子量分布、必需氨基酸组成和在运动表现中的功能作用,以期为运动营养类产品的应用开发提供一定参考和理论支持。     

海蜇低分子肽制备及体外抗氧化活性

目的 优化海蜇低分子肽(low molecular weight peptide from Rhopilema esculentum Kishinouye, RKLP)制备工艺,评价其抗氧化活性及分析氨基酸组成。方法 海蜇利用碱性和中性蛋白酶双酶分步酶解制备海蜇肽,以1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基清除率为指标,响应面实验优化酶解工艺条件。采用超滤技术,获得分子量≤3.5 kDa RKLP。通过测定DPPH自由基、2,2’-联氮-双-3-乙基苯并噻唑啉-6-磺酸[2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonicacid),ABTS]自由基及超氧阴离子(O2-)自由基的清除率,评价RKLP抗氧化活性。氨基酸分析仪分析RKLP氨基酸组成。结果 酶解制备海蜇抗氧化肽的最佳工艺条件为:酶解温度50℃、3%碱性蛋白酶pH 7.5酶解2 h、1%中性蛋白酶pH 7.0酶解2 h,所得的DPPH自由基清除率为71.52%±0.59%,接近与预测值70.57%。RKLP水解度为6...     

Antioxidant and hypoglycaemic effects of tilapia skin collagen peptide in mice

In this study, tilapia collagen peptide (TCP) was prepared by alcalase hydrolysis of tilapia skin, and the antioxidant and hypoglycaemic effects of TCP were investigated. The results showed that TCP possessed DPPH and ABTS+ free radicals‐scavenging activities. In diabetic mice in which diabetes was induced by injection of alloxan (50 mg kg^?1 bw), high‐dose TCP (1.7 g kg^?1 bw) and the drug metformin (1.0 g kg^?1 bw) were found to reduce 31.8% and 30.3% of blood glucose levels in 25 days, respectively. Moreover, in diabetic mice receiving high‐dose TCP, antioxidant enzymes SOD and CAT were increased by 23% and 59.2%, respectively, and MDA was decreased by 39.1%. Comparing the treated high‐dose TCP group with the metformin group, there were similar SOD (61.5 U mg^?1 vs. 60.2 U mg^?1) and MDA (1.4 nmol mg^?1 vs. 1.3 nmol mg^?1), but more (~7%) CAT (359.8 U g^?1 vs. 336.1 U g^?1). Together, the present data, for the first time, demonstrated that TCP possessed hypoglycaemic effects in mice.     

菜籽肽抗氧化活性的研究

用分光光度法测定大鼠红细胞溶血程度,用丙二醛(MDA)试剂盒测定MDA。结果显示,通过腹膜注射菜籽肽,可以使小鼠血清中MDA含量显著减少。高剂量组[100 mg/(kg.d)]与对照组相比,MDA的降低达到极显著水平(P<0.01)。离体实验结果显示,菜籽肽粗品及各级分对体外温育和H2O2诱导的小鼠肝组织匀浆MDA生成有较强的抑制作用,其中粗品RSP-R和级分RSP-1的作用最好,且有量效关系,而对Fe2 诱导的MDA生成的抑制作用稍差。另外,RSP-R还可抑制大鼠红细胞氧化溶血程度。以上结果表明,菜籽肽在体内外均具有明显的抗氧化作用。     

Angiotensin‐I converting enzyme inhibitory and antioxidant activities of peptide fractions extracted by ultrafiltration of cowpea Vigna unguiculata hydrolysates

BACKGROUND: Enzymatic proteolysis of food proteins is used to produce peptide fractions with the potential to act as physiological modulators. Fractionation of these proteins by ultrafiltration results in fractions rich in small peptides with the potential to act as functional food ingredients. The present study investigated the angiotensin‐I converting enzyme (ACE‐I) inhibitory and antioxidant activities for hydrolysates produced by hydrolyzing Vigna unguiculata protein extract as well as ultrafiltered peptide fractions from these hydrolysates. RESULTS: Alcalase^®, Flavourzyme^® and pepsin–pancreatin were used to produce extensively hydrolyzed V. unguiculata protein extract. Degree of hydrolysis (DH) differed between the enzymatic systems and ranged from 35.7% to 58.8%. Fractionation increased in vitro biological activities in the peptide fractions, with IC₅₀ (hydrolysate concentration in µg protein mL^?1 required to produce 50% ACE inhibition) value ranges of 24.3–123 (Alcalase hydrolysate, AH), 0.04–170.6 (Flavourzyme hydrolysate; FH) and 44.7–112 (pepsin–pancreatin hydrolysate, PPH) µg mL^?1, and TEAC (Trolox equivalent antioxidant coefficient) value ranges of 303.2–1457 (AH), 357.4–10 211 (FH) and 267.1–2830.4 (PPH) mmol L^?1 mg^?1 protein. CONCLUSION: The results indicate the possibility of obtaining bioactive peptides from V. unguiculata proteins by means of a controlled protein hydrolysis using Alcalase^®, Flavourzyme^® and pepsin–pancreatin. The V. unguiculata protein hydrolysates and their corresponding ultrafiltered peptide fractions might be utilized for physiologically functional foods with antihypertensive and antioxidant activities. Copyright © 2010 Society of Chemical Industry     

Biological screening of 100 plant extracts for cosmetic use (I): inhibitory activities of tyrosinase and DOPA auto-oxidation

The aim of this study was to evaluate several plant extracts with a view to developing melanogenesis inhibitors. In this study, 100 plant extracts were screened to elucidate their whitening effects using in vitro inhibition of tyrosinase and DOPA auto-oxidation activity. Several plant extracts such as Chaenomeles speciosa, Dryopteris crassirhizoma, Gastrodia ellata, Glycyrrhiza glabra, Morus alba, Myristica fragrans, Rheum palmatum and Sophora japonica showed inhibition of mushroom tyrosinase activity. Plant extracts including Bupleurum falcatum, Caragana sinica, Morus alba and Tussilago farfara showed inhibition of DOPA auto-oxidation activity.