发芽和提取条件对玉米胚芽中超氧化物歧化酶的诱导作用

为提高玉米超氧化物歧化酶(SOD)的活力和提取率,在不同条件下对玉米进行发芽处理,从发芽玉米中剥离胚芽,并在磷酸缓冲液中浸泡,再提取SOD,并测定SOD总活力。结果表明：玉米在30℃、有光照条件下发芽4d,剥离的40g胚芽在0.05mol/L 200mL磷酸缓冲液中浸泡36h,通过胶体磨研磨浸提1h,60℃热沉淀15min,用1.5倍－20℃的丙酮沉淀SOD,经Cellulose-DE-52层析,再经SephadexG-75层析,得到的玉米SOD比活力最高,为4487.28U/mg pro,比未发芽的玉米胚芽提取的SOD的比活力(924.18U/mg pro)提高了3.86倍。     

改良的黄嘌呤氧化酶法测定动植物组织中SOD比活力

针对黄嘌呤氧化酶法测定动植物组织中SOD活力存在干扰的问题,对方法加以改良。以使SOD变性而不影响干扰物质的干扰值为原则,通过测定酶活力及圆二色光谱寻找制备样品空白的最佳实验条件,设计经过加热的样品空白;利用扣除法排除样品中其他物质对黄嘌呤氧化酶体系的干扰;通过加标回收实验以及对动植物组织的实测,对改良前后的测定结果进行比较。结果表明：用改良的黄嘌呤氧化酶法可使动植物组织中SOD活力的检测结果更准确。     

Antioxidant and Hepatoprotective Effects of the Red Ginseng Essential Oil in H(2)O(2)-Treated HepG2 Cells and CCl(4)-Treated Mice

The aim of this study was to evaluate the antioxidant mechanisms of red ginseng essential oil (REO) in cells as well as in an animal model. REO was prepared by a supercritical CO(2) extraction of waste-products generated after hot water extraction of red ginseng. In HepG2 cells, REO diminished the H(2)O(2)-mediated oxidative stress and also restored both the activity and expression of antioxidant enzymes such as superoxide dismutase, catalase and glutathione peroxidase. Administration of REO inhibited the phosphorylation of upstream mitogen-activated protein kinases (MAPKs) such as c-Jun N-terminal kinase, extracellular signal-regulated kinase, and p38. In mice, the CCl(4)-mediated elevation of serum aspartate transaminase and alanine transaminase as well as the induction of hepatic lipid peroxidation were decreased by REO administration. REO treatments also resulted in up-regulation of the antioxidant enzyme expression in the liver. Moreover, increased phosphorylations of MAPKs were inhibited after REO administration. Overall, REO seems to protect the liver from oxidative stress through the activation and induction of antioxidant enzymes via inhibition of MAPKs pathways.     

Cytoprotective effects of geraniin against peroxynitrite- and peroxyl radical-induced cell death via free radical scavenging activity

The role of natural antioxidants in combating the deleterious effects of free radicals has received much attention. In the present study, the cytoprotective effects and the free radical scavenging activity of geraniin, a hydrolysable ellagitannin from Nephelium lappaceum rind, were evaluated by using various approaches. Addition of geraniin to the culture media resulted in a profound cytoprotective effect against damages induced by the peroxynitrite generator 3-morpholinosydnonimine (SIN-1), and the peroxyl radical generator 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH). Geraniin exhibits more potent cytoprotective activity than that of 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox). Geraniin exhibited potent antioxidant activity against reactive species, such as nitric oxide, superoxide anion, and chemically synthesized peroxynitrite. Kinetic analysis of reactivity against peroxyl radicals generated by AAPH revealed that geraniin possesses potent reactivity against peroxyl radicals with higher stoichiometric number than Trolox. The cytoprotective effect of geraniin was only observed when geraniin and these toxic compounds were co-existing, suggesting that geraninn exhibits cytoprotective effects via free radical scavenging activity in the extracellular fluid.     

Production, purification and characterisation of polysaccharides from Pleurotus ostreatus with antitumour activity

BACKGROUND: Mushroom polysaccharides play an important role in functional foods because they exhibit biological modulator properties such as antitumour, antiviral and antibacterial activities. The present study involved the production, purification and characterisation of intracellular and extracellular free and protein‐bound polysaccharides from Pleurotus ostreatus and the investigation of their growth‐inhibitory effect on human carcinoma cell lines. RESULTS: Several fermentation parameters were obtained: batch polysaccharide productivities of 0.013 ± 8.12 × 10^?5 and 0.037 ± 0.0005 g L^?1 day^?1 for intracellular and extracellular polysaccharides respectively, a maximum biomass concentration of 9.35 ± 0.18 g L^?1, P_max = 0.935 ± 0.018 g L^?1 day^?1, µ_max = 0.218 ± 0.02 day^?1, Y_EP/X = 0.040 ± 0.0015 g g^?1 and Y_IP/X = 0.014 ± 0.0003 g g^?1. Some polysaccharides exhibited superoxide dismutase (SOD)‐like activity of 50‐200 units. Fourier transform infrared analysis of the polysaccharides revealed absorption bands characteristic of such biological macromolecules. Cytotoxicity assays showed that both intracellular and extracellular polysaccharides exhibited antitumour activity towards several tested human carcinoma cell lines in a dose‐dependent manner. CONCLUSION: The polysaccharides of P. ostreatus exhibited high SOD‐like activity, which strongly supports their biological effect on tumour cell lines. The extracellular polysaccharides presented the highest antitumour activity towards the RL95 carcinoma cell line and should be further investigated as an antitumour agent. Copyright © 2012 Society of Chemical Industry     

猪肝超氧化物歧化酶的分离纯化与初步表征

研究猪肝SOD的分离提取、纯化以及初步表征。以猪肝为原料,通过抽提液加热、丙酮沉淀和Sephadex G-200层析柱色谱提取、纯化Cu,Zn-SOD;在单因素试验的基础上,采用L(934)设计对提取工艺进行系统优化。结果表明:经过热变性,可以除去大部分杂蛋白,提高了SOD的比活力;最佳的提取条件是:在1.4%的盐浓度下,于70℃水浴中热变性10 min,然后加1.6倍体积的丙酮获得SOD粗沉淀;溶解后的粗酶液经Sephadex G-200层析柱纯化后,经SDS-PAGE电泳检测为单一条带,HPLC检测显示为单一对称峰,均表明最终得到了高纯度的SOD;紫外-可见吸收光谱和ICP-MS分析结果均表明得到的是含铜、锌的SOD,即Cu,Zn-SOD;以邻苯三酚为底物,该酶的Km值为1.574,Vmax为0.268。     

超声波辅助提取甘薯叶中SOD的工艺研究

本文运用超声波细胞破碎仪,对浆液进行超声波处理,并采用Design-Expert软件的中心组合设计方法设计响应面试验,建立了数学模型,通过进行验证实验,得到了最优的提取工艺条件为液固比为28:1、超声时间6 s、次数40次、时间间隔2 s。在此条件下,SOD的活性为359.54 U/g鲜重,与模型预测值的比较误差为1.01%。     

γ辐照对枯草芽孢杆菌营养体的损伤

选用不同剂量γ射线辐照枯草芽孢杆菌营养体,分别用细胞计数、黄嘌呤氧化及脉冲场凝胶电泳法分析了辐照后的细胞存活率、胞内SOD活性及细胞DNA双链断裂水平。研究发现,随着γ辐照吸收剂量的增大,细胞存活率不断下降;SOD活性随剂量的变化无明显的规律;DNA双链断裂水平与细胞存活率密切相关,DNA的释放百分比和断裂水平值随辐照剂量增加而不断增大。结果表明：γ辐照对枯草芽孢杆菌营养体有较高的灭活能力,其损伤效果可能与SOD活性及双链断裂相关。     

利用啤酒酵母生产SOD的提取条件研究

研究了利用异丙醇破壁、丙酮二次纯化提取SOD生产工艺,最佳提取条件为:异丙醇浓度为90%、时间120min、pH7.0,粗酶液的收率为73%,所得SOD的酶比活为3048.7u/mg.与胞内SOD提取老工艺相比,具有工艺简单、设备少、操作简便、成本低等特点.(孙悟)