人血清蛋白- 葡聚糖偶合抗原的制备与表征

将葡聚糖用NaIO4 氧化成带醛基的产物,再与人血清蛋白(HSA)中的氨基反应形成不稳定的席夫碱,在还原剂硼氰氢化钠(NaCNBH3)的作用下,成为稳定的拟糖蛋白抗原,并对影响偶合反应的几个方面进行讨论,包括：不同分子质量的葡聚糖、葡聚糖氧化所用NaIO4 的量、氧化葡聚糖与HSA 的比例和偶合时间。结果表明：偶合反应的最佳条件为：葡聚糖T40 与氧化剂NaIO4 物质的量比为1:187,偶合溶液pH9.0,氧化葡聚糖T40 与HSA物质的量比为4:1,偶合时间 12h。质量相等的不同类型葡聚糖,葡聚糖分子质量越大,氧化时需要的氧化剂越多,氧化度越高接枝度也越高。     

Specific release of Albutensins from bovine and human serum albumin

Bovine Albutensin (BA) and human Albutensin (HA) represent f(208–216) of bovine serum albumin (BSA) and f(210–218) of human serum albumin (HSA), respectively. The release of BA and HA was studied during tryptic digestion of serum albumins in solutions at different pH values. Specific release of either BA or HA was achieved by tryptic hydrolysis of serum albumins in 0.1 m NaH₂PO₄ solution adjusted to pH 3.5. Under these conditions, about 29–50% of the theoretical maximum of BA and 18–23% of the theoretical maximum of HA were released at high purity after 18 h digestion. Heat treatment of BSA (90 °C for 1–60 min) prior to tryptic hydrolysis (pH 3.5) did not improve the release of Albutensins. Under the same heating conditions, Albutensin from either BSA or HSA proved to be stable. Tryptic hydrolysates at pH 3.5 of serum albumins were submitted to simulated gastro-intestinal digestion. BA and HA were stable under peptic hydrolysis whilst they were almost completely hydrolysed by Corolase PP™.     

荧光光谱法探究pH值对咖啡酸β-苯乙醇酯与人血清蛋白相互作用的影响

为探究pH值对咖啡酸β-苯乙醇酯（caffeic acid phenethyl ester,CAPE）与人血清蛋白（human serum albumin,HSA）相互作用的影响,采用荧光发射光谱和同步荧光光谱研究在不同pH值（3.0、5.0、7.4）条件下CAPE对HSA的荧光猝灭作用,并计算猝灭常数、结合常数和热力学参数,分析荧光猝灭机理,最后通过位点Marker实验确定结合位点。荧光发射光谱表明,不同pH值条件下CAPE对HSA内源荧光均产生了猝灭作用,生理pH 7.4时猝灭率最高,结合常数最大;热力学参数表明CAPE与HSA主要通过范德华力和氢键作用结合;同步荧光表明CAPE和pH值的共同影响改变了HSA的构象;位点Marker实验表明,CAPE与HSA的结合位点位于Sudlow’s site I附近。结果表明,pH值的增加促进了CAPE与HSA的结合,这将为CAPE活性稳态化保持、靶向递送提供理论依据。     

Foaming Properties of Whey Protein Isolate and λ‐Carrageenan Mixed Systems

Heating protein with polysaccharide under neutral or near neutral pH can induce the formation of soluble complex with improved functional properties. The objective of our research was to investigate the effects of λ‐carrageenan (λC) concentrations and pH on foaming properties of heated whey protein isolate (WPI) and λC soluble complex (h‐cpx) in comparison to heated WPI with added λC (pWPI‐λC), and unheated WPI with λC (WPI‐λC). In all 3 WPI‐λC systems at pH 7, increasing λC concentration led to improved foamability until a certain concentration before it decreased. Despite their higher viscosity, both heated systems (pWPI‐λC and h‐cpx) showed significantly better foamability and foam stability compared to WPI‐λC. Rheological results of foams with 0.25% λC suggested that higher elasticity and viscous films were produced in h‐cpx and pWPI‐λC systems corresponding to better foam stability. Foam microstructure images indicated that foams produced from h‐cpx had thicker film and consisted of smaller initial bubble area and more uniform bubble size. Results from the effect of pH (6.2, 6.5, and 7.0) further confirmed that stronger interactions between WPI and λC during heating contributed to the improved foaming properties. Foam stability was higher in h‐cpx system at all 3 pH levels, especially under pH 6.2 where there were strongest interactions between the biopolymers.     

超声波辅助水酶法提取巴塘核桃油工艺优化及其氧化稳定性

为确定超声波辅助水酶法(果胶酶、纤维素酶、中性蛋白酶)提取巴塘核桃油的最佳工艺及不同储藏条件对核桃油氧化稳定性的影响。通过单因素实验以及L₉(34)正交试验研究pH、酶解温度、酶解时间和加酶量对油脂提取率的影响,得到最佳提取工艺,并以过氧化值和酸价为指标研究该核桃油在不同温度、光照、容器材料以及抗氧化剂下的稳定性。结果表明:巴塘核桃油提取的最佳工艺为pH7、酶解温度45 ℃、酶解时间3.5 h、加酶量1.4%,在此条件下巴塘核桃油提取率为78.91%±0.03%;将此条件下提取的核桃油用铁罐灌装,置于冷藏、避光的条件下,并添加0.02% BHT+V_C(质量比1:1)作为抗氧化剂,其表现出的氧化稳定性较佳,具有较长的保质期。     

抗坏血酸降解机理以及脱氢抗坏血酸对食品的影响研究进展

L-抗坏血酸（L-ascorbic Acid，AA）在食品中既能发挥抗氧化、抗褐变的作用，又能在食品体系中发生降解氧化产生褐色色素。因此正确认识其发生降解的机理，保护AA在食品中稳定性至关重要。该研究比较详细地综述了AA在不同条件下（氧、温度、pH值、底物浓度）的自降解机理以及参与美拉德反应的降解过程，同时对抗坏血酸降解产物-脱氢抗坏血酸（Dehydroascorbic Acid，DHAA）在食品中的有利和不利影响进行了总结，一方面DHAA可与面筋巯基交联，提高面团筋性；而另一方面，DHAA又会导致酚类物质降解，导致食品抗氧化能力降低；并且DHAA能够与蛋白质形成糖基化终产物，对人体产生有害物质。该综述以期为AA在食品加工中的降解提供更全面的研究视角。     

Interaction of anthocyanins with human serum albumin: Influence of pH and chemical structure on binding

The affinity of anthocyanins for human serum albumin (HSA) was determined by a fluorescence quenching method. The effects of pH and structure of anthocyanins on the binding constants were studied. The constants for binding of anthocyanins to HSA ranged from 1.08 × 10⁵ to 13.2 × 10⁵ M⁻¹. A hydrophobic effect at acidic pH was shown by the relatively high positive entropy values under the conditions studied. Electrostatic interactions, including hydrogen bonding, contributed to the binding at pH 7.4. The effect of structure of anthocyanins on the affinity was pH-dependent, particularly the effect of additional hydroxyl substituents. Hydroxyl substituents and glycosylation of anthocyanins decreased the affinity for binding to HSA at lower pH (especially pH 4), but increased the strength of binding at pH 7.4. In contrast, methylation of a hydroxyl group enhanced the binding at acidic pH, whilst this substitution reduced the strength of binding at pH 7.4. This paper shows that changes in anthocyanin structure or reductions in pH, which may occur in the region of inflammatory sites, have an effect on the binding of anthocyanins to HSA.     

外源抗氧化剂α-生育酚在河蟹肌肉脂肪-蛋白质氧化体系中的作用

为了考察添加α-生育酚对河蟹肌肉中氧化脂肪-蛋白质体系的影响,以采集的河蟹肉为研究对象,通过测定蟹肉肌原纤维蛋白中巯基含量、表面疏水性、二级结构变化以及蛋白电泳情况,研究常温贮藏条件下氧化脂肪-蛋白质模拟体系中肌原纤维蛋白的变化。结果显示,蟹油经过12、24?h氧化后,脂肪酸组成发生变化,不饱和脂肪酸相对含量明显下降。添加24?h氧化蟹油的河蟹肌肉中,活性巯基含量的减少和表面疏水性的增加较为显著,氧化程度越高,α-螺旋、β-折叠相对含量越少,β-转角则越多;添加α-生育酚的处理组与相应的对照组相比,活性巯基含量的减少、表面疏水性的增加显著减缓,α-螺旋、β-折叠相对含量较多,β-转角较少,十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（sodium dodecyl sulfate-polyacrylamide gel electrophoresis,SDS-PAGE）结果显示,抗氧化剂的添加对肌原纤维蛋白降解有一定的抑制效果,条带光密度分析显示仅有个别组间条带有显著差异,SDS-PAGE用于检测蛋白质氧化程度不及活性巯基和表面疏水性指标灵敏。研究表明氧化蟹油能够加速蛋白质的变性,且氧化程度越高,效果越显著;α-生育酚对氧化脂肪诱导的蛋白质变性具有显著的抑制作用。     

Antioxidant properties of humic substances

Humic substances (HS) are heterogeneous, redox-active organic macromolecules. While electron transfer to and from HS under reducing conditions is well investigated, comparatively little is known on the electron donating (i.e., antioxidant) properties of HS under oxic conditions. In this work, the electron donating capacities (EDCs) of terrestrial and aquatic HS were quantified by mediated electrochemical oxidation over a wide range of pH values and applied redox potentials (E(h)) using 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) as an electron transfer mediator. Electrochemical oxidation of three model humic acids (HAs) was largely irreversible, and the EDCs of these HAs increased with increasing E(h) and pH. These results suggest that HS contain a wide variety of moieties that are oxidized at different potentials and that, upon oxidation, release protons and undergo irreversible follow-up reactions. At a given pH and E(h), the EDCs of the HS correlated well with their titrated phenol contents suggesting phenolic moieties as major electron donating groups in HS. Comparing the EDCs of 15 HS with their electron accepting capacities (EACs), aquatic HS had higher EDCs and lower EACs than terrestrial HS of comparable aromaticities. These results indicate that oxidative transformation of HS in the environment results in a depletion of electron donating phenolic moieties with antioxidant properties relative to the electron accepting quinone moieties.     

Binding of citrus flavanones and their glucuronides and chalcones to human serum albumin

Naringenin and hesperetin glycosides are the major polyphenols (flavanones) of citrus fruits and juices and are thought to participate in the cardioprotective effects of diets rich in plant products. Naringenin and hesperetin glucuronides (resulting from conjugation at the A- or B-ring) are the main circulating metabolites in humans and their binding to human serum albumin (HSA) is expected to modulate their half-life in plasma and tissue distribution. In this work, the binding of flavanone glucuronides to HSA was investigated by fluorescence spectroscopy. Binding constants in the range of 3-9 × 10(4) M(-1) were estimated. The affinity of glucuronides for HSA is close to that of naringenin and hesperetin themselves. Competition experiments in the presence of the fluorescent probes dansylsarcosine and quercetin were used to gain information on the flavanone binding site. Naringenin and hesperetin chalcones were also included for comparison as their glucuronides too were detected in the general circulation. Naringenin and hesperetin chalcones spontaneously undergo cyclization back to the parent flavanones under neutral conditions. The cyclization was significantly slowed down by HSA but led to a racemic mixture of (2R) and (2S) flavanones in the absence or presence of HSA.     

Preparation,antioxidant activity and protective effect of coconut testa oil extraction on oxidative damage to human serum albumin

Extraction, physicochemical properties and fatty acid composition analysis of coconut testa oil (CTO), antioxidant activity and the protective effect on oxidative damage to human serum albumin (HSA) of coconut testa oil extract (CTOE) were investigated. Results showed that the optimal extract condition of CTO was B₃A₂C₂ (temperature of 60 °C, material‐to‐solvent of 1:4 g mL^?1 and extraction time of 3 h) with the maximum oil yield (76.83 ± 0.53%). The obtained CTO was nondrying oil with iodine value of 14.69 g per 100 g, and lauric acid was the main component of 42.28%. Hydrogen peroxide scavenging activity of CTOE can reach to 49.81% at 2.5 mg mL^?1, while antioxidant activity (AA) on the oxidation of linoleic acid dropped from 56.82% to 31.70% during the first 80 min. CTOE could prevent HSA from oxidative damage induced by hydrogen peroxide via inhibiting the formation of protein carbonyl and increase of hydroperoxides content effectively. Total phenolic content was 68 mg g^?1, and the epicatechin and catechin were 2.74 and 2.26 mg g^?1 in its phenolic compositions, respectively. These all suggested CTO and CTOE might be new worthy exploiting functional sources.     

Limited hydrolysis of β‐casein by cell wall proteinase and its effect on hydrolysates's conformational and structural properties

Optimisation of enzymatic hydrolysis of β‐casein with cell envelope proteinase (CEP) from Lactobacillus acidophilus JQ‐1 to produce the angiotensin‐I‐converting enzyme (ACE) inhibitory peptides using response surface methodology (RSM). Under optimal conditions (enzyme‐to‐substrate ([E]/[S]) ratio (w/w) of 0.132 and pH of 8.00 at 38.8 °C), the ACE inhibitory activity of hydrolysates was 72.06% and the total peptides was 11.75 mg mL^?1. Scanning electron microscopy (SEM) micrographs indicated that the tightness of the β‐casein surface structure was gradually weakened and small holes appeared after enzymatic treatment, while Fourier transform infrared spectroscopy (FTIR) spectra indicated remarkable changes in the chemical composition and macromolecular conformation of β‐casein after enzymatic hydrolysis. Differential scanning calorimetry (DSC) analysis indicated that the corresponding hydrolysates had higher thermal stability. The enzymatic hydrolysis also led to an increase in the free sulfhydryl content of β‐casein hydrolysates compared with raw β‐casein, which led to the increase in the antioxidant activity of β‐casein hydrolysates.     

石榴皮提取物与焦磷酸钠复配对猪肉饼品质的影响

为探究焦磷酸钠(sodium pyrophosphate,TSPP)、异抗坏血酸钠(sodium D-isoascorbate,D-V_CNa)与石榴皮提取物(pomegranate peel extract,PPE)复配使用在肉制品中是否具有协同增效作用,以丁基羟基茴香醚(butyl hydroyanisole,BHA)为阳性对照,通过对冷藏期间猪肉饼的pH、蒸煮损失、色差、质构特性、脂肪氧化(硫代巴比妥酸值)和蛋白氧化(总巯基、蛋白羰基含量)的表征,研究各抗氧化剂单独或复配使用对猪肉饼品质及贮藏氧化稳定性的影响。结果表明,TSPP可以提高猪肉饼的pH,而D-V_CNa与PPE的添加对肉饼的pH无显著影响;TSPP的添加可以降低猪肉饼的蒸煮损失,改善肉饼的质构特性,且效果优于PPE、BHA。冷藏过程中肉饼不断发生脂肪和蛋白氧化,其中TSPP对脂肪氧化的抑制效果与BHA相近,但对蛋白氧化无显著抑制效果; PPE具有良好的抗氧化性能,可以明显抑制猪肉饼的脂肪氧化和蛋白羰基的生成,且在冷藏前期可以抑制巯基向二硫键的转化。整体来看,PPE+TSPP及P...     

Stability of protein‐stabilised β‐carotene nanodispersions against heating,salts and pH

BACKGROUND: Milk proteins are used in a wide range of formulated food emulsions. The stability of food emulsions depends on their ingredients and processing conditions. In this work, β‐carotene nanodispersions were prepared with selected milk‐protein products using solvent‐displacement method. The objective of this work was to evaluate the stability of these nanodispersions against heating, salts and pH. RESULTS: Sodium caseinate (SC)‐stabilised nanodispersions possessed the smallest mean particle size of 17 nm, while those prepared with whey‐protein products resulted in larger mean particle sizes (45–127 nm). Formation of large particles (mean particle size of 300 nm) started after 1 h of heating at 60 °C in nanodispersions prepared with SC. More drastic particle size changes were observed in nanodispersions prepared with whey protein concentrate and whey protein isolate. The SC‐stabilised nanodispersions were fairly stable against Na⁺ ions at concentrations below 100 mmol L^?1, but drastic aggregation occurred in ≥ 50 mmol L^?1 CaCl₂ solutions. Aggregation was also observed in whey protein‐stabilised nanodispersions after the addition of NaCl and CaCl₂ solutions. All sample exhibited the smallest mean particle size at neutral pH, but large aggregates were formed at both ends of extreme pH and at pH around the isoelectric point of the proteins. CONCLUSION: The nanodispersions prepared with SC were generally more stable against thermal processing, ionic strength and pH, compared to those prepared with whey proteins. The stable β‐carotene nanodispersions showed a good potential for industrial applications. Copyright © 2008 Society of Chemical Industry     

Effects of artificial lighting on bioactivity of sweet red pepper (Capsicum annuum L.)

Recent studies have shown that artificial light effectively promotes the growth and development of red peppers when light spectra, intensities and duration are controlled. Cited reports deal mostly with studies involving with the growth stages of pepper. Changes in the biochemical status of plants were not studied. Therefore, this study focuses on the changes in the content of some bioactive compounds (polyphenols, flavonoids, tannins, ascorbic acid and antioxidant capacities) after artificial lightening [high‐pressure sodium and light‐emitting plasma (LEP) lamps] during the cultivation of sweet red pepper. The bioactive compounds differ slightly, depending on the source of light. Fluorescence spectra showed higher binding properties of LEP polyphenol extracts to human serum albumin (HSA) than other samples. FTIR peaks were similar in their polyphenols region for all investigated samples.     

Effect of reaction pH and CuSO4 addition on the formation of catechinone due to oxidation of (+)‐catechin

A novel hair dyeing technique being milder and safer for a human body is desired. The oxidation product of (+)‐catechin, catechinone, was invented as a safer dyestuff for hair colouring under such the situation. The preparation of catechinone by a chemical oxidation is a practical way and the objective of the study is clarify the effect of the solution pH and in the presence or absence of Cu²⁺ on the formation rate and yield of catechinone in order to improve the efficiency of the dye formation. The catechinone formation was monitored by ultraviolet‐visible spectroscopy. Catechinone was prepared chemically from (+)‐catechin in aqueous solution with O₂ gas introduced over a pH range of 7.1–11.7. The rate and amount of the dye formation increase with increasing pH. Dissociation of the hydroxyl group of the catechol part of (+)‐catechin is significant for the oxidation of (+)‐catechin and promotes the dye production. This is because the deprotonated (+)‐catechin has a higher reactivity with O₂. The production of catechinone is accelerated by the addition of CuSO₄ and the production rate reaches the maximum at pH = 8.8. (+)‐Catechin ‐ Cu²⁺ complexes are formed and the formation promotes the oxidation of the catechol part of (+)‐catechin at pH ≤ 8.8. On the other hand, the complex becomes too stable to proceed for the oxidation reaction at pH > 8.8.     

两步水解法制备玉米-大豆复合抗氧化肽及其稳定性研究

采用麦芽粉和碱性蛋白酶Alcalase两步水解玉米醇溶蛋白和大豆分离蛋白的混合物(质量比1∶1)制备复合抗氧化肽,以水解度、可溶性蛋白含量和抗氧化活性为指标,对第二步Alcalase的水解条件进行优化,确定两步水解法制备复合抗氧化肽的最适条件。另外,对复合抗氧化肽相对分子质量分布、稳定性和氨基酸组成进行了研究。结果表明,制备复合抗氧化肽的最适条件为在底物质量分数10%、pH 5.5、50℃、酶(麦芽粉)底质量比30%条件下水解3 h后,再在底物质量分数10%、pH 8.5、60℃、酶(Alcalase)底质量比0.75%条件下继续水解3 h。在最适条件下,水解度、可溶性蛋白含量、羟自由基清除率和亚铁离子螯合率分别为36.32%、36.83 mg/mL、40.01%和86.83%,相对分子质量主要分布在300~6 500 Da (含量92.44%)范围内,必需氨基酸指数(EAAI)值为8.02(n=9),属于优质蛋白源。复合抗氧化肽具有热稳定性,在中性及碱性条件下稳定,但不耐受酸性(pH 3~5)、4℃贮存和反复冻融的加工条件。     

Cranberry proanthocyanidins associate with low‐density lipoprotein and inhibit in vitro Cu2+‐induced oxidation

Antioxidant activity of six fractions of cranberry phenolic compounds was determined by inhibition of Cu²⁺‐induced low‐density lipoprotein (LDL) oxidation. The phenolic composition of each fraction was determined by high‐performance liquid chromatography. The phenolic fractions were mixed with aliquots of modified human serum prior to LDL isolation. The serum was modified to remove very‐low‐density lipoprotein and chylomicrons that may bind phenolic compounds. Only fractions 5 and 6 that contained proanthocyanidins (PAs) significantly increased the lag time of LDL oxidation, and the lag time for fraction 6 was significantly higher than for fraction 5. The mass distribution of PAs in these fractions was obtained by matrix‐assisted laser desorption/ionisation time‐of‐flight mass spectrometry, a technique that allows rapid characterisation of the molecular weight distribution in mixtures of oligomeric compounds. Fraction 5 contained trimers through heptamers, whereas fraction 6 contained pentamers through nonamers. In addition, fraction 6 contained PA oligomers with more doubly linked, A‐type interflavan bonds. Results indicate that PAs specifically associate with LDL in modified serum and increase the lag time of Cu²⁺‐induced oxidation. Differences between fractions 5 and 6 in PA structure and effects on LDL oxidation suggest that the degree of polymerisation and the nature of the interflavan bond influence antioxidant properties. © 2001 Society of Chemical Industry     

Thermal Stability of Some Metabolically Conjugated Potential Precursors of Flavor Components in Meat and Milk

The stability of selected metabolic conjugates (phenylphosphate, phenylglucuronide, naphthylglucuronide, and naphthylsulfate) was evaluated in model systems. Phenylphosphate was less stable than the glucuronides and the sulfate, especially under neutral (pH 6.8) and acidic conditions (pH 1.5). Phenylphosphate was completely degraded after 1.7 hr at 100°C, and after 0.3 hr at 150°C at pH 6.8. The glucuronide and sulfate conjugates were especially stable under neutral conditions (pH 6.8) requiring > 5 hr for complete hydrolysis at 100°C. All conjugates were quite stable to hydrolysis under alkaline conditions (pH 11). The presence of certain organic solvents (ethyl acetate or ethyl ether) caused rapid hydrolysis of conjugates under very mild conditions (40°C for 30 min). This information may improve under-standing of flavor production from animal products.     

提取条件对大豆11S 球蛋白得率及纯度的影响

以低脂质含量豆粕为原料,探讨大豆11S球蛋白分级分离过程中提取条件(pH值、还原剂和冷沉时间)对蛋白得率及纯度的影响。结果表明：将浸提液pH值由6.2调至6.6时,大豆11s球蛋白的得率降低了16.71%,而纯度提高了4.60%。随着浸提pH值的提高,蛋白质的巯基与二硫键含量增加。与二硫苏糖醇(DTT)或β-巯基乙醇(ME)相比,在pH6.4的浸提液中添加亚硫酸氢钠(SBS),大豆11s球蛋白的得率最高。而DTT、ME和SBS还原剂中,使用DTT时的11S球蛋白组分得率与纯度最低。在蛋白沉淀阶段,当冷沉时间为10h时,蛋白的得率与纯度最佳,随着冷沉时间的延长(12～16h)巯基与二硫键总量并无显著差异(P≥0.05),而部分巯基发生氧化形成二硫键,大豆11S球蛋白游离巯基含量降低。