锰过氧化物酶(MnP)的研究进展

综述了国内外锰过氧化物酶（MnP)研究的最新动态。MnP是一种含有血红素的过氧化物酶,仅在白腐菌中发现。从产生MnP的微生物、MnP对Mn^2 的依赖性、催化机制、蛋白质研究进展、基因结构和基因表达的研究以及工业应用前景方面进行了系统概述。     

锰氧化细菌的生理生态功能与作用机制研究进展

锰的生物地球化学循环过程与全球尺度的营养元素循环紧密联系,是影响全球生态平衡及气候变化的重要因素之一。在自然界中,锰元素主要以氧化锰和含氧酸盐的矿物形式存在,近年来的研究观点普遍认为细菌介导的氧化作用是自然环境中锰氧化物形成的主要原因。锰氧化细菌广泛分布于海洋、锰矿土壤等生态系统,近期在植物微生态系统中也被发现,其生理生态功能未知。细菌的锰氧化过程是一个复杂的过程,多铜氧化酶和过氧化物(氢)酶是参与该过程的主要酶类,但关于其催化机制的认识尚不成熟。本综述系统探讨锰氧化细菌的种类和分布、细菌锰氧化作用的生理生态功能、参与细菌锰氧化作用的功能酶及其分子机制,总结这一研究领域所取得的成果和仍未解决的科学问题,并对今后的发展方向进行展望。     

植物谷胱甘肽过氧化物酶研究进展

氧化胁迫可诱导植物多种防御酶的产生,其中包括超氧化物歧化酶(SOD,EC1.15.L1)、抗坏血酸过氧化物酶(APX,EC1.11.1.11)、过氧化氢酶(CAT,E.C.1.11.1.6)和谷胱甘肽过氧化物酶(GPXs,EC1.11.1.9).它们在清除活性氧过程中起着不同的作用.GPXs是动物体内清除氧自由基的主要酶类,但它在植物中的功能报道甚少.最近几年研究表明,植物体内也存在类似于哺乳动物的GPXs家族,并对其功能研究已初见端倪.本文综述了有关GPXs的结构以及植物GPXs功能的研究进展.     

植物抗坏血酸过氧化物酶研究进展(综述)

本文从酶的作用机制、酶学特征、分子生物学等方面综述植物抗坏血酸过氧化物酶（APX）的研究进展。     

植物谷胱甘肽过氧化物酶研究进展

氧化胁迫可诱导植物多种防御酶的产生, 其中包括超氧化物歧化酶(SOD, EC1.15.1.1)、抗坏血酸过氧化物酶(APX, EC1.11.1.11)、过氧化氢酶(CAT, E.C.1.11.1.6 )和谷胱甘肽过氧化物酶(GPXs,EC1.11.1.9)。它们在清除活性氧过程中起着不同的作用。GPXs是动物体内清除氧自由基的主要酶类,但它在植物中的功能报道甚少。最近几年研究表明, 植物体内也存在类似于哺乳动物的GPXs家族, 并对其功能研究已初见端倪。本文综述了有关GPXs的结构以及植物GPXs功能的研究进展。     

过氧化物酶体脂肪酸β氧化

除线粒体外,过氧化物酶体也是真核细胞脂肪酸β氧化分解的重要部位．过氧化物酶体β氧化过程包括氧化、加水、脱氢和硫解4步反应,主要参与极长链、支链脂肪酸等的分解．近年关于过氧化物酶体β氧化的研究活跃,在代谢途径及功能等方面有了新的认识,尤其在对相关代谢酶的研究中取得了较大进展．本文就过氧化物酶体β氧化相关进展作一综述．     

感染束顶病后香蕉过氧化物酶和多酚氧化酶活性变化(简报)

香蕉束顶病株的过氧化物酶（POD）和多酚氧化酶（PPO）的活性变化均呈单峰曲线,两者在香蕉束顶病毒（BBTV）浸染前期被诱导活性显著提高,接种后21和28d最高,42d后则都比健株低。侵染前期BBTV在体内的运转受抑,接种叶在接种后28d病毒含量最高,而顶叶则在35d后为最高。     

多功能过氧化物酶介导Mn(III)络合体系对酚类化合物的氧化降解

【背景】酚类化合物是环境中主要的水体污染物之一。多功能过氧化物酶(versatile peroxidase,VP)介导的Mn (III)-有机酸络合体系具有较高的氧化还原电势,在酚类有机污染物降解方面具有巨大潜力。【目的】探究VP介导的Mn (III)-有机酸络合体系降解酚类化合物的能力,为酚类化合物的降解提供新的思路和方法。【方法】研究选取了糙皮侧耳(Pleurotus eryngii)来源的多功能过氧化物酶(PeVP),采用包涵体复性的方法获得了PeVP活性蛋白,并对重组PeVP进行酶学性质研究及Mn (III)络合体系反应条件优化,进而探究Mn (III)络合体系对酚类污染物的氧化降解能力。【结果】确定了PeVP包涵体复性最佳条件为：pH 9.5、10%甘油、0.5 mol/L尿素、0.5 mmol/L氧化型谷胱甘肽(glutathione oxidized,GSSG)、0.1 mmol/L二硫苏糖醇(dithiothreitol,DTT)、0.1 mmol/L乙二胺四乙酸(ethylenediamine tetraacetic acid,EDTA)、5 mmol/L CaCl₂、5µmol/L羟高铁血红素(hematin),4℃静置透析24 h,最后5µmol/L hematin孵育12 h。通过对PeVP介导的Mn (III)-有机酸络合体系优化,确定了最优反应条件为：75 mmol/L苹果酸缓冲液(pH 4.5)、6 mmol/L Mn²⁺和0.2 mmol/L H₂O₂。在上述条件下,探究了络合体系对2,2-丁香醛连氮-双-3-乙基苯并噻唑啉-6-磺酸[2,2ʹ-azinobis-(3-ethylbenzthiazoline-6-sulphonate),ABTS]、2,6-二甲氧基苯酚(2,6-dimethoxyphenol,DMP)、愈创木酚和丁香醛连氮4种酚类模式底物的催化活性,发现在pH 4.5条件下,络合体系对酚类模式底物的氧化活性可达到PeVP直接氧化活性的1.5−7.5倍,并且在16 h的酶解过程中,苯酚、对苯二酚、间苯二酚和对硝基苯酚的平均降解速率分别为10.91、10.69、6.50和5.71 mg/(L·h),推测Mn (III)-有机酸络合物对酚类底物的氧化降解是通过夺取酚类底物的电子形成酚类自由基中间体,自由基中间体经过电子重排和C−C键的断裂,最终导致酚类物质的氧化降解。【结论】在弱酸(pH 4.5)条件下PeVP介导的Mn (III)-苹果酸络合体系对酚类污染物具备较强的氧化能力,这为酚类有机污染物提供了新的生物解决方案。     

辣根过氧化物酶的结构与作用机制

辣根过氧化物酶是一种重要的酶制剂,它已经有一个多世纪的研究历史了。近几年,有关它的结构、催化中间体、催化机制以及特殊氨基酸残基功能等又有了新的发现。     

一株锰氧化细菌的分离、鉴定及其锰氧化特性

【目的】获得锰氧化细菌,对锰矿周边土壤中生物所参与的锰氧化过程进行初探。【方法】依据细菌是否能氧化Mn(Ⅱ),形成棕褐色锰氧化物进行筛选。利用染料LBB对生成的锰氧化物进行检测。通过考察分离菌株的形态、生理特征和16S r RNA基因、gyr B基因、gyr A基因序列的同源性对分离菌株进行鉴定。分析筛选菌与所在属已知锰氧化菌的亲缘关系。利用LBB显色法检测氧化锰的动态生成,通过扫描电镜-能谱分析和X射线衍射技术分析生物氧化锰的表征。【结果】获得1株锰氧化细菌菌株,命名为CP133,综合形态、生理及分子分析结果,鉴定为蜡样芽孢杆菌(Bacillus cereus),分离菌株与多株分离自海洋及土壤的芽孢类锰氧化菌在进化上具有一定的差异。与其他菌株比较菌株CP133具有较强的锰氧化能力,进入稳定期后可生成紧密结合在菌体周围的无定形态生物氧化锰。【结论】从锰矿周边土壤分离出1株具有较强锰氧化功能的蜡样芽孢杆菌,丰富了土壤芽孢类锰氧化菌的资源,同时也为锰矿周围土壤与锰氧化菌间的生物地球化学循环提供了线索及材料。     

Characterization of the solution reactivity of a basic heme peroxidase from Cucumis sativus

A basic heme peroxidase has been isolated from cucumber (Cucumis sativus) peelings and characterized through electronic and (1)H NMR spectra from pH 3 to 11. The protein, as isolated, contains a high-spin ferriheme which in the low pH region is sensitive to two acid-base equilibria with apparent pK(a) values of approximately 5 and 3.6, assigned to the distal histidine and to a heme propionate, respectively. At high pH, a new low-spin species develops with an apparent pK(a) of 11, likely due to the binding of an hydroxide ion to the sixth (axial) coordination position of the Fe(III). A number of acid-base equilibria involving heme propionates and residues in the distal cavity also affect the binding of inorganic anions such as cyanide, azide, and fluoride to the ferriheme, as well as the catalytic activity. The reduction potentials of the native protein and of its cyanide derivative, determined through UV-Vis spectroelectrochemistry, result to be -0.320+/-0.015 and -0.412+/-0.010V, respectively. Overall, the reactivity of this protein parallels those of other plant peroxidases, especially horseradish peroxidase. However, some differences exist in the acid-base equilibria affecting its reactivity and in the reduction potential, likely as a result of small structural differences in the heme distal and proximal cavities.     

Studies on the superoxide anion production and peroxidase activity in potato leaf cell suspension exposed to salicylic acid

It is now widely accepted that salicylic acid (SA) signaling is mediated by reactive oxygen species (ROS) production. We have studied the effect of SA on peroxidase activity and superoxide anion production in potato leaf cell suspension. The results show that potato cells are insensitive to low concentrations of exogenous SA (< 1 mM) and the effect is observed at 1–5 mM SA. The cells exposed to SA exhibit higher peroxidase activity and show different peroxidase pattern when analyzed on native gels compared to the control. Superoxide anion production is enhanced after two hours of treatment and 2.5 mM SA gives the highest value. The results suggest peroxidase-mediated detoxification of ROS elicited by SA.     

Screening, identification and kinetic characterization of a bacterium for Mn(II) uptake and oxidation

Following sample collection and screening at a number of Mn-associated mine sites in Northern Australia, a microbial strain was selected for its enhanced rate of Mn uptake. The strain was identified by phylogenetic analysis as a Rhizobium sp. Kinetic studies of Mn(II) uptake and oxidation by this strain in glucose-based media established that the uptake of Mn(II) was much greater than the conversion of Mn(II) to Mn oxide. Chemical analysis and scanning electron microscopy confirmed the production of significant amounts of polysaccharides by this strain. These polysaccharides may play a role both in enhancing Mn(II) accumulation and in minimizing Mn oxide production.     

Microbial Populations of the River-Recharged Fredericton Aquifer

The City of Fredericton, New Brunswick, Canada obtains its water from the Saint John River-recharged Fredericton aquifer. Water quality improves as it flows from the river into the aquifer in a process called bank filtration. Microorganisms contribute to water quality improvements during bank filtration by removing organic carbon. In the Fredericton aquifer, microbially catalyzed reductive dissolution of manganese oxide minerals negatively affects water quality. Aerobic and anaerobic microorganisms were cultured from Fredericton aquifer production well water, from associated bedrock groundwater, and from Saint John River sediment core and sediment groundwater samples. Aerobes were the largest culturable groups in all samples. The ratio of aerobes to other microbial groups, i.e., those reducing nitrate, Mn^{4 +}, Fe^{3 +}, or sulfate, did not vary significantly along the water flow path from the river to the aquifer. Analysis of microbial community composition along the flow path indicated an essentially identical community except in the immediate vicinity of the aquifer. This is in agreement with the absence of macroscale redox zones in the sediment below the Saint John River as determined by groundwater geochemical data. Bacteria isolated from groundwater samples, identified by 16S ribosomal RNA gene sequencing, were α -, β -, γ -, and δ -Proteobacteria, Actinobacteria, and Firmicutes. In contrast to groundwater samples, the majority of bacteria isolated from sediments were γ -Proteobacteria. Numbers of manganese-reducing bacteria, including Aeromonas spp., were small, however Mn^{4 +} reduction ability was widespread in bacteria enriched and isolated with other electron acceptors. The diverse Fredericton aquifer microbial community likely uses manganese oxide minerals as a sink for electrons derived from organic carbon oxidation.