The scavenging of free radical and oxygen species activities and hydration capacity of collagen hydrolysates from walleye pollock (Theragra chalcogramma) skin

Fish skin collagen hydrolysates (FSCH) were prepared from walleye pollock (Theragra chalcogramma) using a mixture of enzymes, namely trypsin and flavourzyme. The degree of hydrolysis of the skin collagen was 27.3%. FSCH was mainly composed of low-molecular-weight peptides and the relative proportion of <1000Da fraction was 70.6%. Free radical and oxygen species scavenging activities of FSCH were investigated in four model systems, including diphenylpicrylhy-drazyl radical (DPPH), superoxide anion radical, hydroxyl radical and hydrogen peroxide model, and compared with that of a native antioxidant, reduced glutathione (GSH). FSCH was also evaluated by water-absorbing and water-holding capacity. The results showed that FSCH was able to scavenge free radical and oxygen species significantly and to enhance water-absorbing and water-holding capacity remarkably. Therefore, FSCH may have potential applications in the medicine and food industries.     

Effects of polypeptide from Chlamys farreri on amino acid con-tent in guinea pig skin irradiated by chronic ultraviolet A and B

1 INTRODUCTION Long-term exposure of the skin to ultraviolet (UV) radiation results in degenerative processes involved in photoaging and photocarcinogenesis (Differy, 1987). As UVC is absorbed entirely by the stratospheric ozone layer and does not reach the earth抯 surface, sunlight is composed of UVA (320 nm 400 nm) and UVB (290 nm 320 nm). Human skin is exposed constantly to both UVA and UVB radiation, it is, therefore, important to protect the skin against UVA and UVB-induce…     

Isolation and Characterization of Collagen from Squid(Ommastreoges bartrami) Skin

Collagen of squid (Ommastrephes bartrami) skin was examined in the present study. Histology showed that collagen fiber in the skin was partially cross-linked with muscle fiber. Acid-solubilized collagen (ASC) and pepsin-solubilized collagen (PSC) were extracted from the skin and characterized. The results of amino acid composition and electrophoretic patterns revealed that ASC and PSC were both type Ⅰ collagen, containing α1 and α2 chains. FTIR (fourier transform infrared spectroscopy) investigations con-firmed the existence of helical arrangements in PSC of squid skin. The denaturation temperature (Td) and shrinkage temperature (Ts)of PSC were 29.4℃ and 52.8℃, respectively.     

Effects of polypeptide from<Emphasis Type="Italic">Chlamys farreri</Emphasis> on amino acid content in guinea pig skin irradiated by chronic ultraviolet A and B

We examined the effects of polypeptide fromChlamys farreri (PCF) on the amount of hydroxyproline in guinea pig skin irradiated by chronic ultraviolet A (UVA) and ultraviolet B (UVB) radiation. PCF was applied locally before repeated exposure of guinea pig to UVA and UVB. The contents of hydroxyproline and other amino acids in guinea pig skin were determined by automatic amino acid analyzer. Our results showed that: (1) long-time UVA and UVB radiation can reduce dramatically the amounts of hydroxyproline, aspartic acid, threonine, glycine, phenylalanine and lysine in guinea pig skin in comparison with the control group (P<0.05); (2) Compared with model group, pre-treatment with 5% and 20% PCF prior to UVA and UVB radiation can inhibit the decline of amino acids content in guinea pig skin in a dose-dependent manner (P<0.05). As the decrease of hydroxyproline, glycine and lysine contents in the skin directly reflexes type I collagen degeneration, our results indicated that the chronic application of PCF can protect skin type I collagen against UV radiation, and thus protect skin from photoaging. Project No. 39970638 supported by the NSFC, and also supported by the Science and Technology Bureau of Qingdao (No: 2001-28-50).     

Isolation and characterization of collagen from squid (Ommastrephes bartrami) skin

Collagen of squid (Ommastrephes bartrami) skin was examined in the present study. Histology showed that collagen fiber in the skin was partially cross-linked with muscle fiber. Acid-solubilized collagen (ASC) and pepsin-solubilized collagen (PSC) were extracted from the skin and characterized. The results of amino acid composition and electrophoretic patterns revealed that ASC and PSC were both type I collagen, containing α1 and α2 chains. FTIR (fourier transform infrared spectroscopy) investigations confirmed the existence of helical arrangements in PSC of squid skin. The denaturation temperature (T_d) and shrinkage temperature (T_s) of PSC were 29.4°C and 52.8°C, respectively.     

Comparative Study on the Antioxidant Activity of Peptides from Pearl Oyster(Pinctada martensii) Mantle Type V Collagen and Tilapia(Oreochromis niloticus) Scale Type Ⅰ Collagen

In this study, Pearl oyster mantle type V collagen(POMC) and tilapia scale type I collagen(TSC) were extracted and hydrolyzed by various proteases in order to obtain peptides. The antioxidant activity of the peptides was investigated by DPPH, hydroxyl radical scavenging experiments and a dynamic digestion model in vitro. The results show that there are significant differences in amino acid composition between POMC and TSC. The collagen peptides obtained from pearl oyster mantle(POMCP) by treating with alkaline protease exhibited higher antioxidant activity than that from tilapia scale(TSCP) treated with papaya protease, and both of them showed greater DPPH and hydroxyl radical scavenging activity than other peptides. After being separated via Sephadex G-25 chromatography, the M1 fraction isolated from POMCP, and the S1 fraction from TSCP with which both had higher molecular weights showed the strongest antioxidant activity than other fractions, and the M1 fraction exhibited stronger antioxidant activity than the S1 fraction in scavenging free-radicals and protecting cells from the oxidation damage. Furthermore, after treating the dynamic digestion system model in vitro, the DPPH and hydroxyl radical scavenging activity of the M1 fraction increased slightly. These results suggest that POMCP exhibits stronger antioxidant activity than TSCP, which means that PMOP may be a good candidate to be a potential natural antioxidant in the food-processing industry.     

Soil physicochemical properties and vegetation structure along an elevation gradient and implications for the response of alpine plant development to climate change on the northern slopes of the Qilian Mountains

Elevation is one of key factors to affect changes in the environment, particularly changes in conditions of light, water and heat. Studying the soil physicochemical properties and vegetation structure along an elevation gradient is important for understanding the responses of alpine plants andtheir growing environment to climate change. In this study, we studied plant coverage, plant height, species richness, soil water-holding capacity, soil organic carbon(SOC) and total nitrogen(N) on the northern slopes of the Qilian Mountains at elevations from2124 to 3665 m. The following conclusions were drawn:(1) With the increase of elevation, plant coverage and species richness first increased and then decreased, with the maximum values being at 3177 m.Plant height was significantly and negatively correlated with elevation(r=–0.97, P0.01), and the ratio of decrease with elevation was 0.82 cm·100 m-1.(2) Both soil water-holding capacity and soil porosity increased on the northern slopes of the Qilian Mountains with the increase of elevation. The soil saturated water content at the 0-40 cm depth first increased and then stabilized with a further increase of elevation, and the average ratio of increase was2.44 mm·100 m-1. With the increase of elevation, the average bulk density at the 0-40 cm depth first decreased and then stabilized at 0.89 g/cm3.(3) With the increase of elevation, the average SOC content at the 0-40 cm depths first increased and then decreased,and the average total N content at the 0-40 cm depth first increased and then stabilized. The correlation between average SOC content and average total N content reached significant level. According to the results of this study, the distribution of plants showed a mono-peak curve with increasing elevation on the northern slopes of the Qilian Mountains. The limiting factor for plant growth at the high elevation areas was not soil physicochemical properties, and therefore,global warming will likely facilitate the development of plant at high elevation areas in the Qilian Mountains.     

Macroalgae in a spring stream in Shanxi Province： composition and relation to physical and chemical variables

Fourteen stream segments were investigated throughout the Xin’an Spring in Shanxi Province, China in 2004. The variation ranges in stream size, current velocity, discharge, dissolved oxygen, and specific conductance were large. Twenty-two macroalgae species were found in the stream. Major divisions in terms of species numbers were Chlorophyta (59.1%), Cyanophyta (22.8%), Xanthophyta (9.1%), Rhodophyta (4.5%) and Charophyta (4.5%). The most widespread species, Cladophora rivularis (50.0%), also Oedogonium sp. (42.9%) and Spirogyra sp. (42.9%) were well represented throughout the stream, whereas another 10 species were found in only one sampling site. Total percentage cover varied from <1% to 90%. Red algae Batrachospermum acuatum and the charophytes Chara vulgaris have the highest percentage cover. Among the parameters analyzed, the stream width, specific conductance and dissolved oxygen were the ones that more closely related to the species number and percentage cover of macroalgal communities. The species number of each site was negatively correlated with dissolved oxygen content. The total percentage cover of the macroalgae was negatively correlated with the stream width and the specific conductance.     

Protective effect of gelatin and gelatin hydrolysate from salmon skin on UV irradiation-induced photoaging of mice skin

The objective of this study was to investigate the effect of gelatin(SG) isolated from salmon skin and its hydrolysate(SGH) on photoaging skin, and the mechanism responsible for anti-photoaging. The average molecular weights of SG and SGH were 65 k Da and 873 Da, respectively. The amino acid compositions of SG and SGH were similar. Both of them were abundant in hydrophobic amino acids. Twenty-five peptides were identified from SGH. SG and SGH could improve UV irradiation-induced pathological changes of macroscopical tissue texture and skin morphology. Hydroxyproline content is an indicator of matrix collagen content, SG and SGH could inhibit the decrease of hydroxyproline content in photoaging skin in a dose dependent manner. In addition, SG and SGH could alleviate UV irradiation-induced oxidative damages to skin by increasing the activities of total superoxide dismutase(T-SOD), glutathione peroxidase(GSH-Px) and catalase(CAT), increasing the content of glutathione(GSH) and decreasing the content of malonaldehyde(MDA). Moreover, SG and SGH could enhance immune regulation system by increasing the thymus index. Thus, the anti-photoaging mechanisms of SG and SGH were by inhibiting the depletion of antioxidant defense components, involving in the synthesis of collagen and enhancing the function of immune system. Besides, SGH showed a better result in protecting skin from photoaging than SG.     

Antioxidative capacity and enzyme activity in Haematococcus pluvialis cells exposed to superoxide free radicals

The antioxidative capacity of astaxanthin and enzyme activity of reactive oxygen eliminating enzymes such as superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX) were studied in three cell types of Haematococcus pluvialis exposed to high concentrations of a superoxide anion radical (O₂⁻). The results show that defensive enzymes and astaxanthin-related mechanisms were both active in H. pluvialis during exposure to reactive oxygen species (ROS) such as O₂⁻. Astaxanthin reacted with ROS much faster than did the protective enzymes, and had the strongest antioxidative capacity to protect against lipid peroxidation. The defensive mechanisms varied significantly between the three cell types and were related to the level of astaxanthin that had accumulated in those cells. Astaxanthin-enriched red cells had the strongest antioxidative capacity, followed by brown cells, and astaxanthin-deficient green cells. Although there was no significant increase in expression of protective enzymes, the malondialdehyde (MDA) content in red cells was sustained at a low level because of the antioxidative effect of astaxanthin, which quenched O₂⁻ before the protective enzymes could act. In green cells, astaxanthin is very low or absent; therefore, scavenging of ROS is inevitably reliant on antioxidative enzymes. Accordingly, in green cells, these enzymes play the leading role in scavenging ROS, and the expression of these enzymes is rapidly increased to reduce excessive ROS. However, because ROS were constantly increased in this study, the enhance enzyme activity in the green cells was not able to repair the ROS damage, leading to elevated MDA content. Of the four defensive enzymes measured in astaxanthin-deficient green cells, SOD eliminates O₂⁻, POD eliminates H₂O₂, which is a by-product of SOD activity, and APX and CAT are then initiated to scavenge excessive ROS.     

The antioxidant effects of complexes of tilapia fish skin collagen and different marine oligosaccharides

An excess of reactive oxygen species (ROS) leads to a variety of chronic health problems.As potent antioxidants, marine bioactive extracts containing oligosaccharides and peptides have been extensively studied.Recently, there is a growing interest in protein-polysaccharide complexes because of their potential uses in pharmaceutical and food industries.However, only few studies are available on the antioxidant activities of such complexes, in terms of their ROS scavenging capability.In this study, we combined different marine oligosaccharides (isolated and purified) with collagen peptides derived from tilapia fish skin, and evaluated the an-tioxidant activity of the marine peptide-oligosaccharide complexes vis-à-vis the activity of their original component molecules.Bio-chemical and cellular assays were performed to measure the scavenging effects on 1, 1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl and superoxide radicals, and to evaluate the influences on the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and the level of malondialdehyde (MDA) in UV-induced photoaging models.The results indicated that the antioxidant activities of all the complexes were stronger than those of their individual components.Among the 11 complexes tested, two complexes, namely MA1000+CP and κ-ca3000+CP, turned out to be highly effective antioxidants.Although the detailed mechanisms of this improved scavenging ability are not fully understood, this work provides insights into the design of highly efficient peptide-oligosaccharide complexes for potential applications in pharmaceutical, cosmetics and food industries.     

Effect of dietary supplementation of vitamin C on growth,reactive oxygen species,and antioxidant enzyme activity of Apostichopus japonicus (Selenka) juveniles exposed to nitrite

Different amounts of vitamin C were added to diets fed to juveniles(2.5 ± 0.15 g) of sea cucumber Apostichopus japonic u s(Selenka) in an attempt to reduce the stress response of specimens exposed to nitrite stress. A commercial feed was used as the control diet and three experimental diets were made by supplementing 1 000, 1 500, or 2 000 mg vitamin C/kg diet to control diet separately in a 45-day experiment. Sea cucumbers were exposed to three different levels(0.5, 1.0, and 1.5 mg/L) of nitrite stress for 4, 8, and 12 h at four time intervals(0, 15, 30, and 45 d). Growth of the animals was recorded during the experiment. Reactive oxygen species(ROS)(i.e. hydroxyl free radical(-OH), malondialdehyde(MDA) and total antioxidant capacity(T-AOC)) and antioxidant enzyme activities(i.e., superoxide dismutase(SOD) and catalase(CAT)) were measured. Response surface methodology(RSM) was used to analyze the effect of multiple factors on ROS indices and enzyme activities. Weight gain(WG) and special growth rate(SGR) of vitamin C supplementation groups were significantly higher than those of control group( P 0.05). The levels of-OH and MDA increased under exposure time extending and nitrite concentration increasing, whereas T-AOC level decreased. SOD and CAT activities increased at 4 h and 8 h and decreased at 12 h. During the days in which the animal consumed experimental diets, the levels of-OH and MDA decreased and that of T-AOC increased. This result suggests that diets containing vitamin C could reduce the nitrite stress response in the animals and increase their antioxidant capacity. The multifactor regression equation of growth performance, ROS indices, and duration of feeding results suggest that vitamin C supplementation of 1 400–2 000 mg/kg diet for 29–35 days could reduce effectively the effects of nitrite exposure.     

Study on the Free Radical Scavenging Activity of Sea Cucumber(Paracaudina chinens var.) Gelatin Hydrolysate

Gelatin from the sea cucumber(Paracaudina chinens var.) was hydrolyzed by bromelain and the hydrolysate was found to have a high free radical scavenging activity. The hydrolysate was fractionated through an ultrafiltration membrane with 5 kDa molecular weight cutoff(MWCO). The portion(less than 5 kDa) was further separated by Sephadex G-25. The active peak was col-lected and assayed for free radical scavenging activity. The scavenging rates for superoxide anion radicals(O2·-) and hydroxyl radi-cals(·OH) of the fraction with the highest activity were 29.02% and 75.41%,respectively. A rabbit liver mitochondrial free radical damage model was adopted to study the free radical scavenging activity of the fraction. The results showed that the sea cucumber gelatin hydrolysate can prevent the damage of rabbit liver and mitochondria.     

Antioxidant and antitumor potential of wild and IMTA-cultivated Osmundea pinnatifida

Osmundea pinnatifida is a red edible seaweed known as pepper dulse. O. pinnatifida was cultivated in the farm of ALGAplus(Ilhavo, Portugal). This farm is integrated with a seabream and seabass commercial aquaculture and uses the nutrient-enriched water resultant from the fish production as its cultivation medium in the integrated multi-trophic aquaculture(IMTA) manner. Wild and IMTA-cultivated samples of O. pinnatifida were screened for antioxidant and antitumor activities. The antioxidant capacity of solvent extracts from wild and IMTA cultivated samples was assessed in two methods(2,2-diphenyl-1-picrylhydrazyl(DPPH) and oxygen radical absorbance capacity(ORAC)), and their total phenolic contents(TPC) were estimated. Antitumor activity was evaluated in three different tumor cell lines(HepG-2, MCF-7, and SH-SY5Y) through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)assay. Among the solvents used for extraction, dichloromethane was the most effective to extract phenolic compounds and presented higher ORAC. A significant correlation was found between TPC and ORAC,which was also sustained by the principal components analysis(PCA). Dichloromethane extracts induced a cytostatic effect on MCF-7 cells and showed weak cytotoxicity to SH-SY5Y cells and weak impact on cell proliferation. Overall, there were no statistically significant differences in the biological activities shown by the wild and IMTA-cultivated samples. Hence, O. pinnatifida can be obtained in an economical and environmentally sustainable way through IMTA, maintaining bioactive properties in a high potential for further nutraceutical purposes.     

罗非鱼鱼鳞胶制备降血压胶原肽的工艺及酶解物活性

以罗非鱼鱼鳞胶为原料,制备对血管紧张素转换酶(ACE)有抑制活性的降血压肽。以水解度(DH)及ACE抑制率为指标,筛选水解鱼鳞胶的最适酶种,并对其水解工艺条件进行优化;采用超滤技术对所制备的降血压肽进行分离纯化,比较不同组分的ACE抑制率,测定高ACE抑制率组分的氨基酸组成,并运用MALDI-TOF-MS法分析其分子质量分布。结果表明,复合蛋白酶是水解鱼鳞胶最适酶种,在50℃、pH7.5、料液质量体积比1∶5、酶与底物质量比(mE/mS)1.5%条件下,酶解6 h,酶解液的ACE抑制作用最强,其水解度DH 73%,ACE的抑制率76.15%。酶解液经超滤分离所获得的分子质量小于2 000 u的组分对ACE抑制率最高,达83.15%;该组分的氨基酸主要含有Gly和Pro,含量分别为22.04%和11.40%;该组分分子质量范围为1 300~2 000 u。     

Protective Effect of Topically Applied Polypeptide from Chlamys farreri Against Ultraviolet Radiation-Induced Chronic Skin Damage in Guinea Pig

INTRODUCTIONSolarultraviolet(UV)radiationisknowntobeamajoretiologicalriskofphotoaging.AsUVC(2 0 0nm -2 90nm)isabsorbedentirelybythestratosphericozonelayer,sunlightontheearthsur faceiscomposedofUVA (3 2 0nm -4 0 0nm)andUVB (2 90nm -3 2 0nm)only.UVAradiationenergyaccountsformorethan 90 %ofsolarUV ;andpenetratesthebasallayerofepidermismoreefficientlythanUVB (Brulsetal.,1984) .However,atsimilardosages,UVBismuchmoremuta genicthanUVA (Drobetskyetal.,1995) .Furthermore ,humanskinisexp…     

不同方法提取马氏珠母贝外套膜胶原蛋白理化性质比较

【目的】系统分析不同方法提取的马氏珠母贝(Pinctada martensii)外套膜胶原蛋白的理化性质及生物学功能。【方法】分别采用酸-酶法和热水法从外套膜中提取得到两种胶原蛋白(A-PSC(Pm)和HSC(Pm)),对其氨基酸组成、微观结构及热变性温度等理化特性进行比较分析。【结果】HSC(Pm)提取率高于A-PSC(Pm);HSC(Pm)中,Gly、Glu与Arg相对含量较高,分别为20.16%、17.57%和10.07%;而A-PSC(Pm)中,Glu、Asp与Arg相对含量为19.67%、11.63%和10.07%;紫外光谱和红外光谱分别显示,两种胶原蛋白均在220 nm左右有强吸收峰,且具有典型的胶原蛋白特征带(酰胺A、B、Ⅰ、Ⅱ和Ⅲ带);扫描电镜结果显示,A-PSC(Pm)呈致密多孔网状结构,HSC(Pm)则基本为薄片状;DSC分析结果表明,HSC(Pm)具有更好的热稳定性。【结论】酸-酶法及热水法提取的胶原蛋白均具有类V型胶原蛋白特性,其结构特征及其理化性质均存在差异,可根据需求选择特定的胶原蛋白提取方法用以开发不同类型的产品。     

Study on the Free Radical Scavenging Activity of Sea Cucumber （Paracaudina chinens vat.） Gelatin Hydrolysate

Gelatin from the sea cucumber （Paracaudina chinens var.） was hydrolyzed by bromelain and the hydrolysate was found to have a high free radical scavenging activity. The hydrolysate was fractionated through an ultrafiltration membrane with 5 kDa molecular weight cutoff （MWCO）. The portion （less than 5 kDa） was further separated by Sephadex G-25. The active peak was collected and assayed for free radical scavenging activity. The scavenging rates for superoxide anion radicals （O2·^-） and hydroxyl radicals （·OH） of the fraction with the highest activity were 29.02% and 75.41%, respectively. A rabbit liver mitochondrial free radical damage model was adopted to study the free radical scavenging activity of the fraction. The results showed that the sea cucumber gelatin hydrolysate can prevent the damage of rabbit liver and mitochondria.     

Positive effect of porphyrans on the lifespan and vitality of Drosophila melanogaster

The effects of degraded porphyran (P1) and natural porphyran (P) on the lifespan and vitality of Drosophila melanogaster are studied. The porphyrans, added daily to the food medium at 0.2% and 1% concentrations, can significantly increase the lifespan in average of 55.79 and 58.23 d in 0.2% P1 diet females and 1% P1 diet males, extending by 12.29% and 8.60% over the corresponding controls, respectively. The effects of porphyrans on D. melanogaster in heat-stress condition were also examined, and found a remarkable increase in survival time. The results which are consistently associated with the use of porphyrans are related to their free radical scavenger action. Considerable increase in vitality demonstrated that vitalities of middle-aged fly (assessed by measuring their mating capacity) was observed after porphyrans addition. Therefore, porphyrans are effective in reducing the rate of aging, and P1 in low molecular weight is better than natural P.     

Protective effect of topically applied polypeptide from Chlamys farreri against ultraviolet radiation-induced chronic skin damage in guinea pig

Polypeptide fromChlamys farreri (PCF), a topical polypeptide isolated fromChlamys farreri, was used in this experiment aimed to investigate the photoprotective effect of PCF against chronic skin damage induced by ultraviolet A (UVA) and ultraviolet B (UVB) radiation. The chronic ultraviolet-irradiated guinea pig model was established, and visible changes in the skin including wrinkling, sagging and erythema were observed. Malondialdehyde (MDA) and antioxidant enzymes including superoxide dismutase (SOD) and glutathione peroxidase (GSH-px) in the dorsal skin were determined using biochemical methods. The results showed: (1) PCF (5 % and 20%) could greatly protect the dorsal skin of guinea pig against wrinkling, sagging and erythema induced by UV radiation in a concentration-dependent manner. (2) PCF could reduce MDA formation in the dorsal skin caused by UV irradiation, while increasing the activities of SOD and GSH-px. (3) The differences among the PCF groups and UV model group were significant (P<0.05,P<0.01). These results indicated that topical application, of PCF provided broad solar UV spectrum photoprotection; and that the antioxidant property of PCF might play a role in photoprotection. Project No. 39970638 supported by the NSFC, and the Science and Technology Bureau of Qingdao (No. 2001-28-50).