用721-分光光度计测定过氧化氢酶活性的新方法

本研究采用两步法对过氧化氢酶活力进行测定。第一步是经典滴定法中的过氧化氢酶与底物作用，第二步采用过氧化物酶与剩余过氧化氢反应，氧供体（DH2）被氧化成棕色物，以棕色深浅反推过氧化氢酶活力单位。结果表明，该方法是一种灵敏、实用的测定过氧化氢酶活性的新方法。     

进出口脱皮花生涉税归类鉴定快速检测方法

建立脱皮花生涉税归类鉴定的快速检测方法——过氧化氢酶和过氧化物酶活性测定法。以pH 7.7的磷酸盐缓冲液提取花生中的过氧化氢酶,加入过氧化氢溶液后30℃水浴15 min,以0.1 mol/L高锰酸钾标准滴定溶液滴定剩余过氧化氢,通过高锰酸钾耗液量计算过氧化氢酶活动度;以pH 7.0的磷酸盐缓冲液提取花生中的过氧化物酶,25℃孵育30 min,加入愈创木酚和过氧化氢,在436 nm波长下,测定初始时和2 min后的吸光度值,计算两者之差ΔA,计算过氧化物酶活力。分别对鲜花生、出口生花生、干燥脱皮花生以及150℃烘烤30 min出口生花生和干燥脱皮花生进行过氧化氢酶和过氧化物酶活性测定。对数据做方差分析,结果有显著性差异。鲜花生中2种酶活性最高,经干燥后活性降低,150℃烘烤30 min后,2种酶失去活性。     

过氧化物酶在食品分子及其胶体体系修饰中的应用

过氧化物酶(peroxidase, POD)是一类大量存在于自然界及生物体内的氧化还原酶。其家族成员众多，但都可以催化以过氧化氢(H2O2)作为底物的氧化反应。现有研究表明可以通过利用过氧化物酶的氧化特性，来提高功能食品的质量属性和营养特性。而辣根过氧化物酶作为过氧化物酶家族的一员，近些年来被大量研究。尤其是在食品研究开发中，有很多都是通过使用辣根过氧化物酶去催化蛋白质或淀粉等食品分子来改善食品本身的性质或获得更合适的食品胶体，因此值得更深层次的挖掘其应用价值及市场。本文在介绍过氧化物酶的结构、催化机理及影响其催化的关键因素基础上，分析了过氧化物酶催化对食品分子结构和性质的影响，综述了近年来过氧化物酶在食品分子及其胶体体系修饰中的一些应用。     

食品中过氧化氢残留快速检测试纸的研制

过氧化氢作为廉价有效的防腐剂、漂白剂普遍被一些食品生产企业超量或非法使用,食品中过氧化氢残留超标会对消费者健康造成巨大威胁.传统的过氧化氢检测方法复杂且耗时较长,为满足现场快速、准确检测过氧化氢残留的需求,本研究利用酶促反应原理,将含有1g/L的四甲基联苯胺(TMB)和10 μg/mL的辣根过氧化物酶(HRP)混合溶液...     

过氧化氢酶的功能及研究

过氧化氢酶是一类广泛存在于动物、植物和微生物体内的氧化酶,其功能是催化细胞内过氧化氢分解,防止氧化。在论述过氧化氢酶类型划分、结构特点及生理功能的基础上,对其分离纯化、活性测定、性质研究做了综述。     

基于聚硫菫的一次性过氧化氢生物传感器的研究

用循环伏安法将硫堇电聚合在丝网印刷碳电极上,再用壳聚糖-二氧化硅溶胶凝胶将辣根过氧化物酶固定于聚硫堇电极表面,利用聚硫堇作为电子传递介体,用壳聚糖-二氧化硅溶胶凝胶固定辣根过氧化物酶并保持酶的生物活性,制成了新型过氧化氢生物传感器。实验发现,该传感器响应快、灵敏度高、稳定性好,对H2O2表现出良好的响应特性;检测线性范围为7.5×10-6～3.12×10-3mol/L,检出限为1.22×10-6mol/L,并具有抗葡萄糖、抗坏血酸等干扰的特点,有望用于食品中过氧化氢残留的检测。     

聚中性红辣根过氧化物酶传感器测定啤酒中的过氧化氢

制备了以电子媒介体聚中性红固定辣根过氧化物酶(HRP)的生物传感器,采用循环伏安法对该传感器的性能进行了研究。结果表明,辣根过氧化物酶在该电极上实现了稳定的直接电子转移反应,传感器在H2O2浓度为3.18×10-8～3.18×10-3mol/L的范围内具有良好的线性相关性,检出限为6.36×10-9mol/L。应用于啤酒中H2O2的测定,回收率为88.5%～98.6%。     

检测牛奶中过氧化氢的电化学酶传感器的研制

构建一种基于聚硫堇的丝网印刷电化学酶传感器,用于牛奶中过氧化氢的检测。将硫堇电聚合在丝网印刷碳电极上,用壳聚糖二氧化硅溶胶凝胶包埋辣根过氧化酶并固定于聚硫堇电极表面,制成新型过氧化氢生物传感器。结果显示：在牛奶标液中加入不同浓度的过氧化氢后,该酶传感器的响应电流与过氧化氢浓度在3×10-5～1.5×10-3mol/L范围内呈良好的线性关系(R＝0.9964),最低检出限为1.675×10-5mol/L。该传感器应用于牛奶中过氧化氢的检测与国标碘量法基本一致,其加样回收率范围为85.6%～89.4%。该酶传感器灵敏快速(15min)、制作方便、样品处理简单,有望用于牛奶中过氧化氢的快速检测。     

用辣根过氧化物酶生物传感器测定啤酒中的过氧化氢

用一种新型的壳聚糖(CS)/聚乙烯吡咯烷酮(PVP)复合膜固定辣根过氧化物酶(HRP),以乙二醛作交联剂,二茂铁(Fc)作媒介体,制备过氧化氢生物传感器。采用循环伏安法对该传感器的性能进行研究。结果表明,传感器在H2O2浓度为3.00×10-8～3.00×10-4 mol/L的范围内具有良好的线性相关性。使用SBA-50生物传感分析仪测定啤酒中的H2O2,回收率为97.6%～101.8%。     

过氧化氢酶

一、在纺织业中应用的过氧化氢酶 过去一些年来，过氧化氢酶在纺织生产中的应用有着日益增长的趋势。它被用予纤维素漂白后过氧化氢的分解。使用过氧化氢酶的好处是，它只攻击过氧化氢。过氧化氢酶分解两摩尔过氧化氢成为两摩尔的水和1摩尔的氧。     

Fermentation of tea in aqueous suspension. Influence of tea peroxidase

The natural peroxidase and catalase of tea leaf were shown to be active under the conditions employed for the fermentation of minced tea leaf in aqueous suspension. Fermentations carried out under nitrogen with ethyl hydrogen peroxide, which is not decomposed by catalase, yielded predominantly thearubigins, a high proportion of which were polymeric in nature. Fermentations using an excess of hydrogen peroxide gave similar results. Fermentations under conditions of controlled dissolved oxygen using both air and hydrogen peroxide resulted in a pigment distribution intermediate between those observed using air alone and using ethyl hydroperoxide under nitrogen. Adjustment of the amount of hydrogen peroxide used to around 120 μmol g^?1 leaf resulted in a pigment distribution closely similar to that of a good quality black tea. A mixture of theaflavins was stable in the presence of air and tea polyphenoloxidase but decomposed rapidly to yield polymeric materials under the action of hydrogen peroxide and peroxidase derived from tea or horseradish. Kinetic measurements suggest that tea flavanols are generally poorer substrates for peroxidase than for catechol oxidase. However, the level of peroxidase is high and, furthermore, in view of the respective pH optima of the two enzymes, its action would be favoured as the pH falls during fermentation. It is postulated that the nature and distribution of the pigments formed during the fermentation of tea is governed in part by the relative actions of catechol oxidase and peroxidase. These actions are, in turn, influenced by the availability of oxygen and the action of catalase.     

Production of peroxidase enzyme by callus cultures of Citrus aurantifolia S

Ammonium sulphate fractionation of citrus flavedo callus showed considerable peroxidase activity with hydrogen peroxide and guaiacol as substrates. The activity of the enzyme was comparable to that of Sigma horseradish peroxidase. Citrus callus may be a potential source of commercial peroxidase enzyme.     

KOJIC ACID CONVERSION TO A YELLOW PRODUCT(S) VIA THE HORSERADISH PEROXIDASE/H2O2 SYSTEM1

Horseradish peroxidase in the presence of hydrogen peroxide oxidizes kojic acid (5-hydroxy-2-hydroxymethyl)-4H-pyran-4-one) to a yellow product(s). The yellow product(s) formed has a major absorbance peak at 375 nm and is fluorescent. The relationships between, and effects of, various concentrations of horseradish peroxidase, kojic acid and hydrogen peroxide on the rate of oxidation of kojic acid to the yellow product(s) are described. The observation that the oxidation of kojic acid to the yellow product(s) occurs best in the presence of very low concentrations of hydrogen peroxide, relative to that of kojic acid, suggests that kojic acid is a poor hydrogen donor (AH₂) for horseradish peroxidase.     

活性氧对河岸葡萄叶圆片抗氧化酶活性的影响

研究了离体处理下活性氧对河岸葡萄叶圆片抗氧化酶活性的影响.结果表明,外源H2O2和Fe2++H2O2处理4 h可引起河岸葡萄叶圆片细胞膜透性明显增加,引起APOD,GPOD和SOD活性明显下降,但低浓度活性氧可不同程度引起叶圆片CAT活性增加;细胞膜透性与活性氧浓度呈显著正相关.     

基于金钯纳米合金修饰的过氧化氢传感器的研制

研制金钯合金纳米粒子修饰的特异性定量检测食品中过氧化氢残留量的过氧化氢传感器。以比表面积大、生物相容性好、具有优良电催化性能的金钯合金纳米粒子固定辣根过氧化物酶于玻碳电极,制得过氧化氢传感器电极。通过循环伏安法及交流阻抗法表征电极在组装过程中的电化学特性,利用计时电流法对传感器性能进行考察。研究表明：该传感器测定H2O2的线性范围为1×10-7～5×10-3mol/L,检测限为8.0×10-7 mol/L,对H2O2具有较好的催化还原活性和良好的检出性能。该传感器制作简单、成本低廉、可重复性强,可用于快速定量检测食品中过氧化氢残留量。     

Horseradish peroxidase degrades lipid hydroperoxides and suppresses lipid peroxidation of polyunsaturated fatty acids in the presence of phenolic antioxidants

Linoleic acid hydroperoxide (LAOOH) was effectively degraded by horseradish peroxidase (HRP) in the presence of quercetin. Several natural phenolic antioxidants, such as quercetin, capsaicin, and alpha-tocopherol, acted as good hydrogen donors in the peroxidase reaction that occurs during lipid hydroperoxide degradation. However, glutathione, which is a non-phenolic antioxidant that acts as a hydrogen donor for glutathione peroxidase, could not suppress lipid peroxidation in the presence of HRP. Lipid hydroperoxides generated from eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were also degraded with HRP in the presence of quercetin, and oxidative decomposition of DHA was suppressed by this reaction.