花生种子铜锌超氧物歧化酶的纯化及理化性质研究

花生种子铜锌超氧物歧化酶经氯仿-乙醇处理、硫酸铵盐析、凝胶过滤和离子交换柱层析等步骤被分离纯化到均一程度。该酶相对分子量为31500Da,由两个相同亚基组成,亚基分子量为16010Da,它在紫外区最大吸收波长为253nm,等电点为4.2,每摩尔酶含2个原子Cu和2个原子Zn,2mmol/L H2O2和0.2mmol/L KCN能完全抑制其活性,该酶由303个氨基酸残基组成,含有较高的丝氨酸和缬氨酸,不含色氨酸和半胱氨酸。纯化后的SOD比活性为5029U/mg,活力回收率为34%。     

Purification and characterization of Cu,Zn superoxide dismutase from pumpkin (Cucurbita moschata) pulp

From ethyl acetate fraction of olive pomace, solid residue from olive oil extraction, a new compound was isolated and purified through solid-phase extraction using silica Cartridge column chromatography. The structure of the new compound was established as a deoxyloganic acid lauryl ester by spectroscopic data including one- and two-dimensional NMR, electrospray ionization mass spectrometry, infrared analysis and UV spectra. The antioxidant activity of the purified compound was evaluated by measuring the radical-scavenging effect on 2,2-diphenyl-1-picrylhydrazyl and by using the ferric reducing antioxidant power assays. The pure compound has not been previously identified in the genus Olea and could be used as for studying the biosynthetic pathway of oleuropein aglycon.     

Fe, Cu and Zn distribution in different components of commercial infant formulas

Fe, Cu and Zn determination by AAS was performed in 35 infant formula samples from different manufacturers. The trace element content was studied in whole, fat and whey milk, and different chemometric tools have been used in order to observe differences or correlations between infant formulas fortified with inorganic salts (iron, copper and zinc sulphates) or with organic and inorganic salts (iron and copper gluconates, or zinc and iron lactates and zinc oxides). Principal-components analysis (PCA) achieved a reduction from nine variables to three (accounting for 80.8% of the total variability), and some differences between infant formula from the two groups were observed.Cluster analysis gave similar results as PCA. Linear discriminant analysis (LDA) allowed the classification of infant formulas in two categories or classes: the first class formed by samples fortified with inorganic salts (category A) and the second one by samples fortified with organic and inorganic salts (category B). The percentages of samples correctly classified were 96.1 and 100.0 for the categories A and B, respectively. After the application of the soft independent modelling of class analogy (SIMCA) approach, percentages of cases correctly assigned of 87.5 and 12.5 were achieved for the categories A and B, respectively. However, it must be said that the application of SIMCA led to bad results because of the small number of sample used, mainly in category B.     

不同消化方法对大蒜中锰、锌、铜、镉含量的影响

目的：研究不同消化方法对大蒜中Mn，Zn，Cu争Cd微量元素含量的影响。方法：采用干法灰化争湿法消化预处理大蒜，干法灰化以时间争温度做单因素实验；湿法消化以酸的比例、放置时间争种类做单因素实验，通过正交实验确定最佳消化条件，利用原子吸收分光光度法测定大蒜中Mn，Zn，Cu争Cd的含量。结果表明：大蒜中Mn，Zn，Cu争Cd的最佳消化条件，干法灰化是Mn450℃，4h；Zn650℃，6h；Cu550℃，4h；Cd 450℃，4h。湿法消化是Mn：HNO3：H2SO4=5l1，16h；Zn：HNO3：HCl=7：1，12h;Cu：HNO3：H2O=1：1，12h；Cd：HNO3：H2SO4=1：2，12h。测定结果的相对标准偏差均在2．0％以下，回收率为99％～101％之间。     

沙棘SOD结构修饰及性质研究

研究沙棘SOD纯化和化学修饰的效果。用沙棘原果为材料,经透析膜进行纯化SOD,选用月桂酸对沙棘SOD进行化学修饰,最后分别对天然SOD和修饰后SOD做热稳定性、抗酶解性、抗酸碱性和金属离子影响的研究。研究结论表明:修饰后SOD的热稳定性与pH稳定性均明显提高。与天然SOD相比,修饰后的蛋白酶酶切耐受性明显增强,同时金属离子对修饰后的SOD影响也较天然SOD不同。     

Effects of copper status on neutrophil function, superoxide dismutase, and copper distribution in steers.

Twelve Holstein steers in a completely randomized block design were fed either a basal diet (concentrate:silage or hay at a DM ratio of 35:65) plus Cu sulfate at 20 ppm of Cu (Cu-supplemented diet) or a basal diet plus ammonium molybdate to obtain 10 ppm of Mo (Cu-depleting diet) on a DM basis in the whole diet for 8 mo. Supplemental Mo was utilized in the Cu-depleting diet to develop a Cu-deficient group. Molybdenum slowly accumulated in the liver in the group fed the Cu-depleting diet. Copper concentrations in the liver and polymorphonuclear neutrophils decreased in the Cu-deficient group compared with the Cu-sufficient group. Plasma Cu concentration did not change during the trial for the Cu-sufficient group. In the Cu-deficient group, plasma Cu concentrations increased during the first 3 mo of the trial, then declined, and remained unchanged for the last 5 mo. Superoxide dismutase activities in red blood cells, polymorphonuclear neutrophils, and whole blood decreased in the Cu-deficient group. Phagocytic capacity was not affected by Cu status, but killing capacity was decreased by low Cu status in the Cu-deficient group by the end of the trial. Glutathione peroxidase activity was unaffected by Cu status. Clinical symptoms of Cu-deficiency were not observed in this trial; there was no evidence of blood hemoglobin or BW gain difference between the two groups. In this study, Cu status affected its distribution in the tissues and related enzyme activities as well as bactericidal function of neutrophils.     

高效液相色谱法测定烟草中的铁、钴、镍、铜、锌、锰

研究了以2-(2-喹啉偶氮)-5-二乙氨基酚(QADEAP)为柱前衍生试剂,甲醇(内含0.2%的醋酸)和pH5.0 HAc-Na Ac溶液梯度洗脱为流动相,Nova-Pak C₁₈液相色谱柱分离,二极管矩阵检测器检测,高效液相色谱法同时测定铁、钴、镍、铜、锌、锰的方法。根据信噪比(S/N=3)各金属离子的检测限分别为:铁8.5 μg/L,钴7.2 μg/L,镍6.4 μg/L,铜5.0 μg/L,锌11μg/L,锰13μg/L,方法相对标准偏差在1.6%~3.8%之间,标准回收率在92%~10 8%,方法用于烟草中痕量铁、钴、镍、铜、锌、锰的测定,结果令人满意。      

硒锌铜交互效应对肉鸡胸肌铜、锌、铁、锰、钙含量的影响

研究了日粮不同硒、锌和铜水平及互作效应对肉鸡胸肌中铜、锌、铁、钙、锰含量的影响.选用1日龄罗斯308肉仔鸡720只,随机分为9个处理组.在玉米-豆粕型日粮基础上添加不同水平的蛋氨酸硒、甘氨酸锌和甘氨酸铜,以饲喂基础日粮组为对照组,试验期7周.结果表明:1)日粮不同硒水平对前中期胸肌铜含量的影响显著,对中期胸肌锌含量影响极显著,对后期胸肌铁含量影响显著,对前中后期胸肌钙含量影响不显著,对前期胸肌锰含量影响显著;2)日粮不同锌水平对前中后期胸肌铜含量影响显著,对中期胸肌锌含量影响极显著,对中期胸肌铁含量影响显著,对前期胸肌锰含量影响显著,对前中后期胸肌钙含量影响不显著;3)日粮不同铜水平对前中期胸肌铜、锌含量的影响显著,对后期胸肌锰含量影响显著,对前中后期胸肌铁、钙含量影响不显著;4)硒、锌和铜的交互作用对前中后期胸肌铜、锌、锰含量影响显著,特别是中期胸肌铜、锌含量影响极显著,对中后期胸肌铁含量影响显著,对中期胸肌钙含量影响显著.     

串叶松香草超氧化物歧化酶提取工艺优化及对D-半乳糖致衰老模型小鼠的保护作用研究

目的 优化串叶松香草超氧化物歧化酶(superoxide dismutase, SOD)的提取工艺, 并研究串叶松香草SOD对D-半乳糖致衰老模型小鼠的保护作用。方法 采用高压脉冲电场法提取SOD, 以串叶松香草为原料, 进行最佳提取工艺的研究, 以SOD酶活力为指标, 在单因素实验基础上, 进一步通过响应面实验对提取工艺进行优化。同时采用随机法把小鼠分为空白对照组、模型组、串叶松香草SOD低、中、高剂量组、维生素C (vitamin C, VC)阳性对照组。除空白对照组外, 其余5组均皮下连续注射D-半乳糖, 采用注射D-半乳糖构建衰老小鼠模型。通过观察和测定各组小鼠的行为和体质量, 研究其衰老的变化情况; 采用分光光度计法测定小鼠血清中SOD、谷胱甘肽过氧化物酶(glutathione peroxidase, GSH-Px)、过氧化氢酶(catalase, CAT)活性和丙二醛(malondialdehyde, MDA)含量。结果 最佳提取工艺条件为: 电场强度20 kV/cm、脉冲数10个、提取时间32 min, 在此条件下, 提取的串叶松香草SOD酶活力为3878.67 U/g。串叶松香草SOD对D-半乳糖致衰老模型小鼠的抗衰老作用表明: 串叶松香草SOD能改善衰老小鼠的体征, 提高衰老小鼠的体质量, 具有较好的抗肝萎缩和脑萎缩作用, 同时还能提高衰老小鼠血清中SOD、GSH-Px、CAT的活力, 降低其MDA含量, 达到延缓衰老的作用。结论 该研究获得了串叶松香草SOD的最佳提取工艺, 得到的串叶松香草SOD能够延缓小鼠衰老。     

黑米中微量营养元素Fe,Zn,Cu和黑米色素的研究

研究黑米中微量元素Fe、Zn、Cu的含量与分布,及光、温度和几种常见金属离子对黑米色素稳定性的影响。原子吸收光谱测定表明,黑米中Fe和Zn的含量分别为38610~(－5)和36010~(－6);主要富集于黑米色素中。Cu的含量均低于1010~(－6)。黑米色素水溶液(pH=3)于温度70~100℃中,随着温度升高和延长加热时间,吸光值明显降低。光照对色素稳定性的影响随时间延长而增大。Fe~(2+)、Fe~(3+)和Sn~(2+)明显的影响色素的稳定性,而Ca~(2+)、Na~+、Cu~(2+)、Zn~(2+)的作用较小,Al~(3+)和Mg~(2+)可提高黑米色素的稳定性。     

Effects of iron and selenium on the production of catalase, superoxide dismutase, and listeriolysin O in Listeria monocytogenes.

Listeria monocytogenes 19112, Scott A, and 10403S were grown in tryptic soy broth (TSB) and TSB supplemented with 25 to 100 microg/ml of iron (Fe) and 0.5 to 2.5 microg/ml selenium (Se) to examine the effects on catalase (CA), superoxide dismutase (SOD), and listeriolysin O (LLO) activities. Growth in TSB supplemented with Fe resulted in significant increases in CA, SOD, and LLO activities in all three strains when compared to growth in TSB. The addition of 0.5 microg/ml Se to TSB resulted in significantly higher CA and LLO activities in L. monocytogenes 19112 but showed no effect on Scott A or 10403S. These results suggest that Fe plays a role in increasing the activities of CA, SOD, and LLO.     

蟒蛇油中铁锰铜锌的测定与去除研究

采用火焰原子吸收光谱法测定了蟒蛇油中Fe、Mn、Cu、Zn等4种金属元素的含量。利用金属离子螯合剂(柠檬酸、抗坏血酸、磷酸二氢钾)螯合萃取去除蟒蛇油中金属元素,抑制蟒蛇油自动氧化作用,以延长蟒蛇油的贮存期、降低其腥臭味。研究结果表明,Fe、Mn、Cu、Zn等4种金属元素的去除率分别为65.07%、26.61%、11.82%、14.76%。金属元素的去除,有效地抑制了蟒蛇油的自动氧化作用,提高了蟒蛇油的氧化稳定性,使蟒蛇油在20℃条件下的预期贮藏时间延长了32~48d。      

Synthesis of superoxide dismutase,catalase and other enzymes and oxygen and superoxide toxicity during changes in oxygen concentration in cultures of brewing yeast

The physiological effects on brewing yeast, growing in semi-defined wort medium, of a sudden transition from aerobiosis to anaerobiosis were studied. Two yeast strains were examined, used for ale and lager fermentations respectively. The reverse transition (from anaerobiosis to aerobiosis) was also examined. Transitions were applied by changing the sparging gas during growth or in stationary phase, and the effects on the specific activities of certain key enzymes and on the viability of the cultures were examined. Neither type of transition led to significant changes in growth rate, the rate of ethanol production or the specific activities of alcohol dehydrogenase and pyruvate decarboxylase. The most significant change was in the specific activity of CuZn-superoxide dismutase, which showed a rapid increase in activity on transition from anaerobiosis to aerobiosis, and a decrease in activity on the reverse transition. Catalase activity in the ale yeast generally followed that of CuZn-superoxide dismutase, whereas in the lager yeast it remained unchanged by the transitions. The transition from anaerobiosis to aerobiosis caused increases in citrate synthase and Mn-superoxide dismutase, though only after a significant lag period. Aerobic to anaerobic transitions caused a decrease in Mn-superoxide dismutase activity, while citrate synthase remained unchanged. Anaerobically grown cells showed a rapid loss in viability on exposure to oxygen (5–7% in the first hour), while aerobically grown cells were unaffected. When anaerobically grown cells were exposed to 0·25 mM -potassium superoxide, there was an 8% loss of viability within 10 min, whereas aerobic cells were not affected. It is concluded that the toxic effect of oxygen is due to superoxide (or a species derived from it) and that the CuZn-superoxide dismutase (but not the Mn-isoenzyme) plays a role in protecting the cells. The de novo synthesis of the CuZn-enzyme is not always rapid enough to confer full protection.     

灵芝多糖对多刺裸腹溞存活、生殖及总SOD活性的影响

采用多刺裸腹溞(Moina mocrocpa)作为实验动物,考察了不同浓度灵芝多糖对多刺裸腹溞寿命、存活率、生殖量和总超氧化物歧化酶(T-SOD)活性的影响.结果表明,灵芝多糖浓度为40～160mg/L时,多糖组多刺裸腹潘的平均寿命及存活率均明显高于对照组,同时对多刺裸腹溞的繁殖率也有显著影响;不同浓度的灵芝多糖均能促进多刺裸腹溞超氧化物歧化酶的活性的提高.结果为多刺裸腹溞作为衰老动物模型的应用提供了理论依据.     

灵芝多糖对多刺裸腹溞存活、生殖及总SOD活性的影响

采用多刺裸腹溞（Moina mocrocpa）作为实验动物,考察了不同浓度灵芝多糖对多刺裸腹溞寿命、存活率、生殖量和总超氧化物歧化酶（T-SOD）活性的影响。结果表明,灵芝多糖浓度为40～160mg/L时,多糖组多刺裸腹溞的平均寿命及存活率均明显高于对照组,同时对多刺裸腹溞的繁殖率也有显著影响;不同浓度的灵芝多糖均能促进多刺裸腹溞超氧化物歧化酶的活性的提高.结果为多刺裸腹溞作为衰老动物模型的应用提供了理论依据。      

火焰原子吸收法测定大豆渣中铁铜锰锌

应用火焰原子吸收分光光度法同时测定大豆渣中铁、铜、锰、锌的含量、结果表明，本方法快速、准确，上述各元素的回收率分别为88．5％、93．3％、103．5％、93．0％。     

Effects of α-lipoic acid supplementation on malondialdehyde contents and superoxide dismutase in rat skeletal muscles

This study investigated the effects of α-lipoic acid (ALA) supplementation on the malondialdehyde (MDA) contents and superoxide dismutase (SOD) protein expression in rat skeletal muscles under non-exercising condition. Fourteen Sprague-Dawley male rats at the age of 6 weeks were randomly divided into 2 groups CS, (normal diet group, n=7) and AS (0.5% ALA supplemented diet group, n=7), and keep non-exercising condition for 4 weeks. The muscle MDA concentration of the AS group was significantly lower than that of the CS group in the both soleus and extensor digitorum longus muscles. The expressions of Cu/Zn-SOD and Mn-SOD proteins in the soleus muscle were significantly higher in the AS group than that in the CS group. From these results, ALA supplementation under non-exercising condition had a role in increasing of antioxidant enzymes in skeletal muscle types of rats.     

植酸酶对蛋鸭锌、铜、镁生物学利用率影响的研究

研究植酸酶对蛋鸭锌、铜、镁生物学利用率的影响.选取200日龄产蛋期金定鸭1 000只,随机分为5个处理,每个处理4个重复,每个重复50只.处理A为对照组,有效磷水平为0.45%;处理B、C、D、E有效磷水平依次为0.38%、0.32%、0.25%、0.18%,同时添加植酸酶500 FTU/kg.结果表明:日粮有效磷水平从0.45%降低到0.25%,同时添加500 FTU/kg植酸酶,不会影响蛋鸭对Ca、P、Cu、Zn的表观利用率及其在蛋鸭血清和胫骨中的含量(P>0.05).当日粮中有效磷水平为0.18%时,即使添加了植酸酶,Ca、P、Cu、Zn的表观利用率及其在蛋鸭血清和胫骨中的含量都会显著降低(P<0.05).降低有效磷水平和添加植酸酶与否不会影响Mg的生物学利用率.添加植酸酶可以降低蛋鸭日粮中的有效磷水平,对Zn、Mg、Cu的生物学利用率无影响,日粮中有效磷水平的适宜范围为0.25%～0.45%.     

茶叶水中重金属铅、铜、锌、锰的浸出率试验研究

以毛尖、普洱茶叶为研究对象,模拟实际生活中泡茶方式,考察了浸泡次数、浸泡时间、浸泡水温3种不同浸泡方式对铜、锌、锰金属元素浸出率的影响,采用火焰原子吸收光谱法检测茶叶中的重金属含量。结果表明:2种茶叶中Pb含量很小,低于检出限,Cu、Zn、Mn的含量未超标,但是Mn含量高,毛尖、普洱茶叶Mn含量分别为696.0mg/kg和659.4mg/kg;浸泡时间为30min、浸泡水温为90℃时,第一次浸泡的茶水中铜、锌、锰的浸出率最高。