Influence of Amino Acid Compositions and Peptide Profiles on Antioxidant Capacities of Two Protein Hydrolysates from Skipjack Tuna (Katsuwonus pelamis) Dark Muscle

Influence of amino acid compositions and peptide profiles on antioxidant capacities of two protein hydrolysates from skipjack tuna (Katsuwonus pelamis) dark muscle was investigated. Dark muscles from skipjack tuna were hydrolyzed using five separate proteases, including pepsin, trypsin, Neutrase, papain and Alcalase. Two hydrolysates, ATH and NTH, prepared using Alcalase and Neutrase, respectively, showed the strongest antioxidant capacities and were further fractionated using ultrafiltration and gel filtration chromatography. Two fractions, Fr.A3 and Fr.B2, isolated from ATH and NTH, respectively, showed strong radical scavenging activities toward 2,2-diphenyl-1-picrylhydrazyl radicals (EC₅₀ 1.08% ± 0.08% and 0.98% ± 0.07%), hydroxyl radicals (EC₅₀ 0.22% ± 0.03% and 0.48% ± 0.05%), and superoxide anion radicals (EC₅₀ 1.31% ± 0.11% and 1.56% ± 1.03%) and effectively inhibited lipid peroxidation. Eighteen peptides from Fr.A3 and 13 peptides from Fr.B2 were isolated by reversed-phase high performance liquid chromatography, and their amino acid sequences were determined. The elevated antioxidant activity of Fr.A3 might be due to its high content of hydrophobic and aromatic amino acid residues (181.1 and 469.9 residues/1000 residues, respectively), small molecular sizes (3–6 peptides), low molecular weights (524.78 kDa), and amino acid sequences (antioxidant score 6.11). This study confirmed that a smaller molecular size, the presence of hydrophobic and aromatic amino acid residues, and the amino acid sequences were the key factors that determined the antioxidant activities of the proteins, hydrolysates and peptides. The results also demonstrated that the derived hydrolysates and fractions from skipjack tuna (K. pelamis) dark muscles could prevent oxidative reactions and might be useful for food preservation and medicinal purposes.     

椰麸组分蛋白的氨基酸组成、结构与乳化性分析

为提高椰子加工副产物的利用,本研究从椰麸中依次提取4种组分蛋白,包括清蛋白、球蛋白、谷蛋白-1和谷蛋白-2.分析这4种组分蛋白的氨基酸组成,并利用SDS-PAGE技术分析其亚基组成和分子量.同时分析其表面疏水性,并研究不同离子强度对椰麸各组分蛋白的乳化性和乳化稳定性的影响.结果表明,椰麸蛋白质中清单白、球蛋白、谷蛋白-...     

Identification of QTL Affecting Protein and Amino Acid Contents in Rice

The phenotypes of protein and amino acid contents were measured in an F9 recombinant inbred line population derived from a cross between Zhenshan 97B and Delong 208. A total of 48 and 64 QTLs were identified in 2004 and 2005, respectively. The contribution of each QTL to the phenotypic variation ranged from 4.0% to 43.7%. Most QTLs co-localized, forming 29 QTL clusters on the chromosomes with three major ones detected in both years, which were mapped on chromosomes 1, 7 and 9, respectively. The two QTL clus...     

Fish Sidestream-Derived Protein Hydrolysates Suppress DSS-Induced Colitis by Modulating Intestinal Inflammation in Mice

Inflammatory bowel disease is characterized by extensive intestinal inflammation, and therapies against the disease target suppression of the inflammatory cascade. Nutrition has been closely linked to the development and suppression of inflammatory bowel disease, which to a large extent is attributed to the complex immunomodulatory properties of nutrients. Diets containing fish have been suggested to promote health and suppress inflammatory diseases. Even though most of the health-promoting properties of fish-derived nutrients are attributed to fish oil, the potential health-promoting properties of fish protein have not been investigated. Fish sidestreams contain large amounts of proteins, currently unexploited, with potential anti-inflammatory properties, and may possess additional benefits through bioactive peptides and free amino acids. In this project, we utilized fish protein hydrolysates, based on mackerel and salmon heads and backbones, as well as flounder skin collagen. Mice fed with a diet supplemented with different fish sidestream-derived protein hydrolysates (5% w/w) were exposed to the model of DSS-induced colitis. The results show that dietary supplements containing protein hydrolysates from salmon heads suppressed chemically-induced colitis development as determined by colon length and pro-inflammatory cytokine production. To evaluate colitis severity, we measured the expression of different pro-inflammatory cytokines and chemokines and found that the same supplement suppressed the pro-inflammatory cytokines IL-6 and TNFα and the chemokines Cxcl1 and Ccl3. We also assessed the levels of the anti-inflammatory cytokines IL-10 and Tgfb and found that selected protein hydrolysates induced their expression. Our findings demonstrate that protein hydrolysates derived from fish sidestreams possess anti-inflammatory properties in the model of DSS-induced colitis, providing a novel underexplored source of health-promoting dietary supplements.     

杂交稻与亲本稻米蛋白质及氨基酸含量的关系

分析了８个三系杂交稻Ｆ１稻米与其相应的５个恢复系和３个不育系及其相应的３个保持系稻米的蛋白质含量和１７种氨基酸含量。结果表明，各组合（系）的蛋白质含量与其氨基酸的绝对含量正相关；杂交稻Ｆ１稻米的蛋白质含量与氨基酸含量高于其相应的恢复系稻米的蛋白质含量和氨基酸含量；不育系稻米的蛋白质含量与氨基酸含量高于其相应保持系稻米的蛋白质含量与氨基酸含量。由此认为，杂交稻稻米蛋白质含量与氨基酸含量受其细胞质基因影响；杂交稻Ｆ１与其三系亲本的蛋白质与氨基酸含量的差异可能是杂交稻表现杂种优势的重要生理基础。     

转AAP1基因玉米游离氨基酸与可溶性蛋白质含量变化关系的研究

氨基酸通透酶1(AAP1)是位于细胞膜上与氨基酸转运有关的载体蛋白。以转组成型启动子(UBI)的AAP1玉米植株(N7-30、N7-26)、转胚乳特异性启动子(Glu)的AAP1玉米植株(KF8-33、KF8-39、KF9-22)为材料,探究转AAP1基因玉米中游离氨基酸和可溶性蛋白质含量之间的变化关系。结果表明,转AAP1基因玉米的根部和叶片中游离氨基酸含量在抽丝期最高;根部可溶性蛋白质含量在成熟期最高;叶片中可溶性蛋白质含量在抽丝期最高。叶片中游离氨基酸含量与可溶性蛋白质含量随着生育期的变化均呈现低-高-低的趋势,抽丝期达到最高值;种子中游离氨基酸的含量与可溶性蛋白质含量呈显著正相关性。转胚乳特异性启动子玉米种子中游离氨基酸含量、脯氨酸含量、赖氨酸含量均显著高于转组成型启动子玉米种子。AAP1可以提高植物对游离氨基酸的吸收与转运,进而提高植物对氮素的利用效率。     

The Skeletal Amino Acid Composition of the Marine Demosponge Aplysina cavernicola

It has been discovered during the past few years that demosponges of the order Verongida such as Aplysina cavernicola exhibit chitin-based skeletons. Verongida sponges are well known to produce bioactive brominated tyrosine derivatives. We could recently demonstrate that brominated compounds do not exclusively occur in the cellular matrix but also in the skeletons of the marine sponges Aplysina cavernicola and Ianthella basta. Our measurements imply that these yet unknown compounds are strongly, possibly covalently bound to the sponge skeletons. In the present work, we determined the skeletal amino acid composition of the demosponge A. cavernicola especially with respect to the presence of halogenated amino acids. The investigations of the skeletons before and after MeOH extraction confirmed that only a small amount of the brominated skeleton-bound compounds dissolves in MeOH. The main part of the brominated compounds is strongly attached to the skeletons but can be extracted for example by using Ba(OH)₂. Various halogenated tyrosine derivatives were identified by GC-MS and LC-MS in these Ba(OH)₂ extracts of the skeletons.     

Physicochemical,Nutritional and In Vitro Antidiabetic Characterisation of Blue Whiting (Micromesistius poutassou) Protein Hydrolysates

Protein hydrolysates from low-value underutilised fish species are potential sources of high-quality dietary protein and health enhancing peptides. Six blue whiting soluble protein hydrolysates (BW-SPH-A_F), generated at industrial scale using different hydrolysis conditions, were assessed in terms of their protein equivalent content, amino acid profile and score and physicochemical properties in addition to their ability to inhibit dipeptidyl peptidase IV (DPP-IV) and stimulate the secretion of insulin from BRIN-BD11 cells. Furthermore, the effect of simulated gastrointestinal digestion (SGID) on the stability of the BW-SPHs and their associated in vitro antidiabetic activity was investigated. The BW-SPHs contained between 70–74% (w/w) protein and all essential and non-essential amino acids. All BW-SPHs mediated DPP-IV inhibitory (IC₅₀: 2.12–2.90 mg protein/mL) and insulin secretory activity (2.5 mg/mL; 4.7 to 6.4-fold increase compared to the basal control (5.6 mM glucose alone)). All BW-SPHs were further hydrolysed during SGID. While the in vitro DPP-IV inhibitory and insulin secretory activity mediated by some BW-SPHs was reduced following SGID, the activity remained high. In general, the insulin secretory activity of the BW-SPHs were 4.5–5.4-fold higher than the basal control following SGID. The BW-SPHs generated herein provide potential for anti-diabetic related functional ingredients, whilst also enhancing environmental and commercial sustainability.     

Physicochemical and Antioxidant Properties of Gelatin and Gelatin Hydrolysates Obtained from Extrusion-Pretreated Fish (Oreochromis sp.) Scales

Fish gelatin and its hydrolysates exhibit a variety of biological characteristics, which include antihypertensive and antioxidant properties. In this study, fish gelatins were extracted from extrusion-pretreated tilapia scales, and then subjected to analyses to determine the physicochemical properties and antioxidant activity of the extracted gelatins. Our findings indicate that TSG2 (preconditioned with 1.26% citric acid) possessed the greatest extraction yield, as well as higher antioxidant activities compared with the other extracted gelatins. Hence, TSG2 was subjected to further hydrolyzation using different proteases and ultrafiltration conditions, which yielded four gelatin hydrolysates: TSGH1, TSGH2, TSGH3, and TSGH4. The results showed that TSGH4 (Pepsin + Pancreatin and ultrafiltration < 3000 Da) had a higher yield and greater antioxidant activity in comparison with the other gelatin hydrolysates. As such, TSGH4 was subjected to further fractionation using a Superdex peptide column and two-stage reverse-phase column HPLC chromatography, yielding a subfraction TSGH4-6-2-b, which possessed the highest 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity compared with the other fractions. Further LC-ESI/MS/MS analysis of TSGH4-6-2-b suggested two novel peptides (GYDEY and EPGKSGEQGAPGEAGAP), which could have potential as naturally-occurring peptides with antioxidant properties. These promising results suggest that these antioxidant peptides could have applications in food products, nutraceuticals, and cosmetics.     

Cosmetic,Biomedical and Pharmaceutical Applications of Fish Gelatin/Hydrolysates

There are several reviews that separately cover different aspects of fish gelatin including its preparation, characteristics, modifications, and applications. Its packaging application in food industry is extensively covered but other applications are not covered or covered alongside with those of collagen. This review is comprehensive, specific to fish gelatin/hydrolysate and cites recent research. It covers cosmetic applications, intrinsic activities, and biomedical applications in wound dressing and wound healing, gene therapy, tissue engineering, implants, and bone substitutes. It also covers its pharmaceutical applications including manufacturing of capsules, coating of microparticles/oils, coating of tablets, stabilization of emulsions and drug delivery (microspheres, nanospheres, scaffolds, microneedles, and hydrogels). The main outcomes are that fish gelatin is immunologically safe, protects from the possibility of transmission of bovine spongiform encephalopathy and foot and mouth diseases, has an economic and environmental benefits, and may be suitable for those that practice religious-based food restrictions, i.e., people of Muslim, Jewish and Hindu faiths. It has unique rheological properties, making it more suitable for certain applications than mammalian gelatins. It can be easily modified to enhance its mechanical properties. However, extensive research is still needed to characterize gelatin hydrolysates, elucidate the Structure Activity Relationship (SAR), and formulate them into dosage forms. Additionally, expansion into cosmetic applications and drug delivery is needed.     

武夷岩茶氨基酸组分及含量分析

采用氨基酸自动分析仪测定分析大红袍、水仙、肉桂、雀舌、金毛猴和矮脚乌龙等6个品种武夷岩茶的游离氨基酸组分,结果表明:6个品种武夷岩茶均含有17种氨基酸,氨基酸含量品种间差异显著,武夷岩茶的游离氨基酸总量为75.82~107.12 mg/g;其中含量较高的为谷氨酸、天冬氨酸和亮氨酸。武夷岩茶的氨基酸中含有各种人体必需氨基酸,且比例均衡;必需氨基酸总量占30.47%~40.46%,药用氨基酸含量丰富,占氨基酸总量的61.29%~63.90%;呈鲜味的氨基酸含量也占氨基酸总量的23.80%~28.63%,因此,武夷岩茶具有较高的营养价值和保健功能。     

Antioxidative Effect of Chlorella Pyrenoidosa Protein Hydrolysates and Their Application in Krill Oil-in-Water Emulsions

Chlorella pyrenoidosa is an excellent source of protein, and in this research, we assessed the antioxidant and emulsifying effects of Chlorella protein hydrolysate (CPH) using neutral proteases and alkaline proteases, as well as the properties of CPH-derived krill oil-in-water (O/W) emulsions. The CPHs exhibited the ability to scavenge several kinds of free radicals, including 1,1-diphenyl-2-picrylhydrazyl (DPPH), O₂⁻, hydroxyl, and ABTS. Additionally, the CPHs (5 mg/mL) scavenged approximately 100% of the DPPH and ABTS. The CPHs showed similar emulsifying activities to Tween 20 and excellent foaming activities (max FS 74%), which helped to stabilize the krill oil-in-water emulsion. Less than 10 mg/mL CPHs was able to form fresh krill oil-in-water emulsions; moreover, the CPHs (5 mg/mL) in a krill O/W emulsion were homogenous, opaque, and stable for at least 30 days. Based on their inhibitory effects on the peroxide value (POV) and thiobarbituric acid reactive substances (TRABS), the CPHs were found to be able to inhibit lipid oxidation in both emulsifying systems and krill O/W emulsions. Thus, the CPHs could improve superoxide dismutase (SOD) activities by 5- or 10-fold and decrease the high reactive oxygen species (ROS) level caused by the addition of H₂O₂ in vitro. In conclusion, health-promoting CPHs could be applied in krill oil-in-water emulsions as both emulsifiers and antioxidants, which could help to improve the oxidative and physical stability of emulsions.     

Ameliorative Effects of Oyster Protein Hydrolysates on Cadmium-Induced Hepatic Injury in Mice

Cadmium (Cd) is a widespread environmental toxicant that can cause severe hepatic injury. Oyster protein hydrolysates (OPs) have potential effects on preventing liver disease. In this study, thirty mice were randomly divided into five groups: the control, Cd, Cd + ethylenediaminetetraacetic acid (EDTA, 100 mg/kg), and low/high dose of OPs-treatment groups (100 mg/kg or 300 mg/kg). After continuous administration for 7 days, the ameliorative effect of OPs on Cd-induced acute hepatic injury in Cd-exposed mice was assessed. The results showed that OPs significantly improved the liver function profiles (serum ALT, AST, LDH, and ALP) in Cd-exposed mice. Histopathological analysis showed that OPs decreased apoptotic bodies, hemorrhage, lymphocyte accumulation, and inflammatory cell infiltration around central veins. OPs significantly retained the activities of SOD, CAT, and GSH-Px, and decreased the elevated hepatic MDA content in Cd-exposed mice. In addition, OPs exhibited a reductive effect on the inflammatory responses (IL-1β, IL-6, and TNF-α) and inhibitory effects on the expression of inflammation-related proteins (MIP-2 and COX-2) and the ERK/NF-κB signaling pathway. OPs suppressed the development of hepatocyte apoptosis (Bax, caspase-3, and Blc-2) and the activation of the PI3K/AKT signaling pathway in Cd-exposed mice. In conclusion, OPs ameliorated the Cd-induced hepatic injury by inhibiting oxidative damage and inflammatory responses, as well as the development of hepatocyte apoptosis via regulating the ERK/NF-κB and PI3K/AKT-related signaling pathways.     

海南鱼酸中乳酸菌的分离鉴定

从海南鱼酸中分离出4株优势乳酸菌L2、L3、L8、L11,根据形态、生理生化特征和分子生物学特征鉴定为植物乳杆菌(Lactobacillus plantarum)。其中L2菌株产酸速率最快,可用于鱼酸纯种发酵。     

不同品质类型小麦籽粒游离氨基酸、蛋白质及其组分的动态变化

为了解小麦氮素营养代谢的动态变化规律,给优质专用小麦品种选育与应用提供科学依据,选用不同品质类型的四个小麦品种(系)济南17、PH82-2-2、PH97-4、PH97-5,在两种施氮水平和两种追肥时期下研究了籽粒游离氨基酸、蛋白质及其组分的动态变化.结果表明,籽粒发育初期游离氨基酸含量高,随籽粒发育其含量逐渐下降;蛋白质含量积累呈现"高-低-高"的变化趋势,品种之间差异显著,成熟期蛋白质含量高的品种一般有较高的游离氨基酸和蛋白质积累水平.不同品质类型小麦籽粒蛋白质组分含量变化动态基本一致,灌浆初期清蛋白含量较高,随籽粒发育逐渐下降,灌浆中后期下降趋缓;球蛋白总体含量水平较低,随籽粒发育缓慢下降,灌浆末期略有上升.醇溶蛋白在籽粒发育前期积累较少,花后14 d快速积累;谷蛋白在花后7 d已有一定的积累,随后其含量逐渐上升,强筋高蛋白品种有较高的醇溶蛋白和谷蛋白积累水平,在成熟籽粒蛋白质中谷蛋白和醇溶蛋白所占比例也高,弱筋低蛋白品种有相对较低的醇溶蛋白和谷蛋白积累,醇溶蛋白和谷蛋白所占比例也低.     

肉桂木栽培6种食用菌氨基酸特性及营养评价

为了评价肉桂木栽培食用菌的营养特性,本研究测定了采用肉桂（Cinnamomum cassia Pres1）生产中废弃物肉桂木栽培的金顶侧耳（Pleurotus citrinopileatus）,秀珍菇（Pleurotus geesteranus）,刺芹侧耳（Pleurotus eryngii）,黑皮鸡枞（Oudemansiella raphanipes）,黄伞（Pholiota adiposa）和猴头菌（Hericium erinaceus）的营养成分及氨基酸组成,通过主成分分析、蛋白质营养价值评价及聚类分析对其蛋白质的营养价值进行评价与分类。结果表明,6种肉桂栽培食用菌的膳食纤维、多糖和蛋白质含量范围分别为24%~46%、4~15 mg/g和19%~43%。主成分1（PC1）和主成分2（PC2）一共解释了92.46%的差异,苏氨酸、亮氨酸、异亮氨酸、苯丙氨酸、缬氨酸共同决定了PC1,赖氨酸决定了PC2。氨基酸评分发现,肉桂木栽培的6种食用菌中,蛋氨酸+半胱氨酸（Met+Cys）含量均过剩,与其他5种食用菌相比,秀珍菇的Met+Cys含量过剩较少,与WHO/FAO模式值相近。聚类分析结果与肉桂木栽培的食用菌的Met+Cys含量有关,黄伞与金顶侧耳距离最近,秀珍菇的Met+Cys含量与其他5种差异较大,聚类距离较远,与WHO/FAO模式值距离较近。综上表明,肉桂木栽培食用菌氨基酸含量丰富,种类比较齐全,是理想的蛋白质来源,但是由于肉桂木栽培食用菌Met+Cys含量偏高,仍需要与其他日常膳食搭配以达到人体氨基酸的平衡供给。     

花生远缘杂交后代的氨基酸含量变异分析

本实验室利用野生花生(A.monticola)与栽培品种(黑花生、桂花红)杂交,获得了一些具有高产性状的花生新材料。通过对这些远缘杂种后代种子的氨基酸含量进行检测,共能检测到17种水解氨基酸和20种游离氨基酸。其中含量最高的是谷氨酸,其次是天冬氨酸和精氨酸,三者的总和占总氨基酸含量的46.1%～47.3%。不同株系间氨基酸含量的变异系数有较大差异。本研究可为利用远缘杂交获得高产优质的花生新材料提供一定的理论依据。     

土壤肥力对小麦开花后各器官游离氨基酸和籽粒蛋白质含量变化的影响

能过随机区组试验和相关的方法，研究了两种土壤肥力条件下小麦开花后植株体内游离氨基酸含量的变化动态及其与籽粒蛋白质含量的关系。结果表明，开花后小麦植株各器官游离氨基酸含量均与籽粒蛋白质含量呈显著或极显著正相关，土壤肥力高能够显著提高各器官中的游离基酸含量，并延长维持高水平的时间，进而促进籽粒蛋白质合成，提高籽闰蛋白质含量和改善品质。     

Amino Acid Profiles and Biopotentiality of Hydrolysates Obtained from Comb Penshell (Atrina pectinata) Viscera Using Subcritical Water Hydrolysis

The recovery of amino acids and other important bioactive compounds from the comb penshell (Atrina pectinata) using subcritical water hydrolysis was performed. A wide range of extraction temperatures from 140 to 290 °C was used to evaluate the release of proteins and amino acids. The amount of crude protein was the highest (36.14 ± 1.39 mg bovine serum albumin/g) at 200 °C, whereas a further increase in temperature showed the degradation of the crude protein content. The highest amount of amino acids (74.80 mg/g) was at 230 °C, indicating that the temperature range of 170–230 °C is suitable for the extraction of protein-rich compounds using subcritical water hydrolysis. Molecular weights of the peptides obtained from comb penshell viscera decreased with the increasing temperature. SDS-PAGE revealed that the molecular weight of peptides present in the hydrolysates above the 200 °C extraction temperature was ≤ 1000 Da. Radical scavenging activities were analyzed to evaluate the antioxidant activities of the hydrolysates. A. pectinata hydrolysates also showed a particularly good antihypertensive activity, proving that this raw material can be an effective source of amino acids and marine bioactive peptides.     

Two Novel Antioxidant Nonapeptides from Protein Hydrolysate of Skate (Raja porosa) Muscle

In the current study, the preparation conditions of neutrase hydrolysate (SMH) from skate (Raja porosa) muscle protein were optimized using orthogonal L₉(3)⁴ tests, and R values indicated that pH was the most important factor affecting HO· scavenging activity of SMH. Under the optimum conditions of pH 7.0, enzymolysis temperature 60 °C, enzyme/substrate ratio (E/S) 2%, and enzymolysis time 5 h, EC₅₀ of SMH on HO· was 2.14 ± 0.17 mg/mL. Using ultrafiltration, gel filtration chromatography, and RP-HPLC, two novel antioxidant nonapeptides (SP-A and SP-B) were isolated from SMH and their amino acid sequences were found to be APPTAYAQS (SP-A) and NWDMEKIWD (SP-B) with calculated molecular masses of 904.98 Da and 1236.38 Da, respectively. Both showed strong antioxidant activities. SP-A and SP-B exhibited good scavenging activities on HO· (EC₅₀ 0.390 and 0.176 mg/mL), DPPH· (EC₅₀ 0.614 and 0.289 mg/mL), and O₂⁻· (EC₅₀ 0.215 and 0.132 mg/mL) in a dose-dependent manner. SP-B was also effective against lipid peroxidation in the model system. The aromatic (2Trp), acidic (2Asp and Glu), and basic (Lys) amino acid residues within the sequences of SP-B might account for its pronounced antioxidant activity. The results of this study suggested that protein hydrolysate and peptides from skate muscle might be effective as food additives for retarding lipid peroxidation occurring in foodstuffs.