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亚硝酸盐氧化还原酶(nitrite oxidoreductase,NXR)是硝化细菌将亚硝酸盐氧化为硝酸盐的关键酶,广泛存在于亚硝酸盐氧化菌中.由于它是可溶性的膜内酶,其催化机理与膜内电子传递密切联系,给它的研究带来了一定的困难.本文综述了多年来国内外研究者从不同方面对NXR研究的成果,详细论述了NXR的组成结构、工作机理以及不同因子对其活性的影响,总结了近几年应用于研究NXR的新方法,并展望了对NXR研究的发展方向及其意义. 相似文献
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本研究主要对克雷伯杆菌甘油转化1,3-丙二醇代谢途径中的2个关键酶甘油脱氢酶(GDH)、1,3-丙二醇氧化还原酶(PDOR)反应机制和动力学进行了研究。首先,通过初速度和产物抑制动力学研究确定了GDH、PDOR双底物酶促反应机制为有序BiBi机制,明确了由反应物消耗到产物生成之间的历程。其次,建立了GDH、PDOR双底物酶促反应动力学模型,由动力学模型可知,在偶合反应中,如果GDH和PDOR酶量相同,GDH氧化反应成为限速反应,而辅酶I将主要以氧化型NAD+形式存在。动力学信息为酶法合成1,3-丙二醇和代谢工程研究提供理论指导。 相似文献
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高等植物叶绿体定位的铁氧还蛋白-NADP+氧化还原酶(LFNR)负责催化光合线性电子传递的最后一步反应,催化电子由还原态的铁氧还蛋白(Fd)传递给NADP+。LFNR分布在叶绿体的3个不同的组分中,即叶绿体基质中、类囊体膜上和叶绿体内膜上。最近的研究表明,大多数膜定位的LFNR并非光合作用所必需的,叶绿体基质中的LFNR足以维持光合作用的正常进行。叶绿体中的两个蛋白——Tic62和TROL作为LFNR的锚定蛋白,可以与LFNR在类囊体膜上形成高分子量的蛋白复合体。Tic62-LFNR复合体主要负责在夜间保护LFNR的活性,但它不直接在光合作用中起作用。然而,TROL-LFNR复合体对植物的光合作用有一定的影响。本文将概述植物LFNR的最新研究进展。 相似文献
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真核生物中正确二硫键的形成是在内质网中由二硫键异构酶PDI及相关蛋白催化的,而在原核生物大肠杆菌中二硫键的氧化、还原和异构化发生在细胞周质,由一系列的二硫键氧化还原酶完成.从1991年Badewell发现第一个氧化还原蛋白DsbA开始,目前已发现有七种二硫键氧化还原酶.DsbB,DsbD、DsbE/CcmG及CcmH位于细胞膜上,DsbA、DsbC,DsbG在细胞周质空间中.DsbA和DsbB的氧化和电子传递链相联系,而DsbC、DsbD,DsbE,DsbG和CcmH的还原需要来自细胞质的电子传递. 相似文献
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高等植物铁氧还蛋白-NADP~+氧化还原酶研究进展 总被引:1,自引:0,他引:1
高等植物叶绿体定位的铁氧还蛋白-NADP+氧化还原酶(LFNR)负责催化光合线性电子传递的最后一步反应,催化电子由还原态的铁氧还蛋白(Fd)传递给NADP+。LFNR分布在叶绿体的3个不同的组分中,即叶绿体基质中、类囊体膜上和叶绿体内膜上。最近的研究表明,大多数膜定位的LFNR并非光合作用所必需的,叶绿体基质中的LFNR足以维持光合作用的正常进行。叶绿体中的两个蛋白——Tic62和TROL作为LFNR的锚定蛋白,可以与LFNR在类囊体膜上形成高分子量的蛋白复合体。Tic62-LFNR复合体主要负责在夜间保护LFNR的活性,但它不直接在光合作用中起作用。然而,TROL-LFNR复合体对植物的光合作用有一定的影响。本文将概述植物LFNR的最新研究进展。 相似文献
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真核生物中正确二硫键的形成是在内质网中由二硫键异构酶PDI及相关蛋白催化的,而在原核生物大肠杆菌中二硫键的氧化、还原和异构化发生在细胞周质,由一系列的二硫键氧化还原酶完成。从1991年Badewell发现第一个氧化还原蛋白DsbA开始,目前已发现有七种二硫键氧化还原酶。DsbB、DsbD、DsbE/CcmG及CcmH位于细胞膜上,DsbA、DsbC、DsbG在细胞周质空间中。DsbA和DsbB的氧化和电子传递链相联系,而DsbC、DsbD、DsbE、DsbG和CcmH的还原需要来自细胞质的电子传递。 相似文献
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土壤水解酶类催化动力学研究进展 总被引:11,自引:3,他引:11
土壤水解酶是存在于土壤中的一种重要的酶类,参与了土壤中为数众多的重要生物化学反应,与土壤中多种营养元素转化密切相关其催化反应的动力学研究常用来阐明其催化过程的特性、酶的本质属性及其对环境变化的响应等,研究土壤水解酶动力学特征对探讨其来源、性质及影响因素,对进一步调控多种营养元素参与的反应过程有着重要意义。文中概述了土壤水解酶的种类及其参与的生物化学反应;探讨了土壤水解酶动力学研究的理论基础;综述了土壤水解酶催化动力学研究的进展和影响因素,在此基础上,对今后研究提出了几点建议。 相似文献
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稻-麦轮作系统土壤水解酶及氧化还原酶活性对开放式空气CO2浓度增高的响应 总被引:9,自引:0,他引:9
研究了FACE条件下(CO2浓度增加200μmol·mol^-1)水稻、小麦不同生育期0~10cm土层土壤脲酶、磷酸酶、芳基硫酸酯酶、脱氢酶活性的变化.结果表明,FACE条件下,土壤脲酶活性在冬小麦生育前期低于对照,在孕穗期高于对照;在水稻生育前期高于对照,在成熟期低于对照.磷酸单酯酶活性在冬小麦生育期高于对照;在水稻分蘖期高于对照,在生育后期(拔节期、抽穗期和成熟期)低于对照.芳基硫酸酯酶活性在小麦越冬期和孕穗期低于对照,在分蘖期和成熟期高于对照;在水稻生育期间均高于对照.脱氢酶活性在小麦和水稻的生育前期低于对照,在后期高于对照. 相似文献
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Yoshimitsu Shimomura Kazuko Sumiguchi‐Agari Ryoji Masui Seiki Kuramitsu Keiichi Fukuyama 《Acta Crystallographica. Section D, Structural Biology》2002,58(8):1365-1367
A probable quinone oxidoreductase (MW = 32.1 kDa) from Thermus thermophilus HB8 was overproduced in Escherichia coli and purified. Gel‐filtration chromatography suggested the protein to be in a dimeric state. This protein enhanced the reduction activity of quinones by NADPH. It was crystallized in the absence and the presence of NADPH by the hanging‐drop vapour‐diffusion method. Both crystals were hexagonal, space group P6122 or P6522, with unit‐cell parameters a = b = 77.6, c = 236.7 Å for the apo form and a = b = 77.6, c = 235.9 Å for the complex with NADPH. They diffract to better than 2.3 Å resolution with synchrotron radiation. The asymmetric unit has one protein subunit (VM = 3.2 Å3 Da−1 and Vsol = 0.62 for the apo form), indicating that the twofold axis of the dimeric protein and the crystallographic twofold axis coincide. 相似文献
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The tungsten- and the molybdenum-containing aldehyde oxidoreductases from Clostridium formicoaceticum show, for aldehydes, K
m values<30 M and K
i values of millimolar concentrations. The tungsten-containing aldehyde oxidoreductase is inactivated to 50% by 3 mM KCN within 1 min, by 1 mM ferricyanide within 5 min, and by 0.05 mM chloralhydrate within 30 s. The molybdenum-containing AOR shows 50% inactivation within 1 min only with 70 mM KCN. The tungsten-containing enzyme is very sensitive to oxygen, especially in the reduced state, whereas the molybdenum-containing enzyme exhibits only moderate oxygen sensitivity without being markedly influenced by the redox state of the enzyme. The tungsten in the aldehyde oxidoreductase is bound to a pterin cofactor (Wco) of the mononucleotide form that is known for molybdopterin cofactor (Moco). The nature of the molybdenum cofactor in the molybdenum-containing aldehyde oxidoreductase is still unclear. The UV/VIS spectrum of the tungsten-containing aldehyde oxidoreductase shows a broad absorption in the range of 400 nm with a millimolar absorption coefficient of 18.1 (reduced form) and 24.8 (dehydrogenated form) at 396 nm. The epr spectrum exhibits two different W(V) signals with the following g values for signal A: 2.035, 1.959, 1.899 and signal B: 2.028, 2.017, 2.002. Dithionite-reduced enzyme shows signals of 4Fe–4S or 2Fe–2S clusters. Initial rate studies with different substrates for the carboxylate reduction led to a Bi Uni Uni Bi mechanism.Abbreviations
AOR
aldehyde oxidoreductase
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NH
2
CO-MV
1,1-carbamoylmethylviologen
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MV
methylviologen
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TMV
1,1,2,2-tetramethylviologen 相似文献
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Azmiri Sultana Igor Alexeev Inari Kursula Pekka Mntsl Jarmo Niemi Gunter Schneider 《Acta Crystallographica. Section D, Structural Biology》2007,63(2):149-159
The crystal structure of aclacinomycin oxidoreductase (AknOx), a tailoring enzyme involved in the biosynthesis of the polyketide antibiotic aclacinomycin, was determined to 1.65 Å resolution by multiwavelength anomalous diffraction using data from selenomethionine‐substituted crystals. The crystals belong to space group P21, with unit‐cell parameters a = 68.2, b = 264.5, c = 68.2 Å, β = 119°. Analysis of the intensity statistics clearly showed the presence of pseudomerohedral twinning. The data set could also be indexed and scaled with an Rsym of 0.072 in the orthorhombic space group C2221 (unit‐cell parameters a = 69.7, b = 117.5, c = 264.4 Å), indicating the possibility of pseudomerohedral twinning along the diagonal between the monoclinic a and c directions. Refinement using this twin operator resulted in an Rfree of 24.2%. A monoclinic lattice with a = c and β close to 120° can emulate a hexagonal metric, with the possibility of a threefold twin operator along the b axis and three twin domains. Refinement assuming three‐domain twinning gave a final Rfree of 26.5%. The structure of AknOx can be thus refined with comparable Rfree values using either of the twin operators separately, suggesting the possibility that crystals of AknOx contain six twin domains generated by the twofold and threefold twin operators perpendicular to each other. Both twin operators coincide with noncrystallographic symmetry axes that may promote twinning. 相似文献
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C. Frazo L. Sieker R. Coelho J. Morais I. Pacheco L. Chen J. LeGall Z. Dauter K. Wilson M. A. Carrondo 《Acta Crystallographica. Section D, Structural Biology》1999,55(8):1465-1467
Crystals of rubredoxin oxygen oxidoreductase have been obtained and characterized. They belong to space group P21212, with unit-cell dimensions a = 88.24 (15), b = 101.25 (7), c = 90.80 (3) Å. The homodimer (86 kDa) in the asymmetric unit is related by a non-crystallographic twofold rotation axis parallel to the ab `diagonal' direction, as shown by the self-rotation maximum in the section with χ = 180°. This pseudo-crystallographic symmetry element was also found to be the twinning axis of pseudo-merohedrally twinned crystals, leading to apparent pseudo-tetragonal P4212 crystal symmetry. 相似文献
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Shankar Prasad Kanaujia Chellamuthu Vasuki Ranjani Jeyaraman Jeyakanthan Masami Nishida Yoshiaki Kitamura Seiki Baba Akio Ebihara Nobutaka Shimizu Noriko Nakagawa Akeo Shinkai Masaki Yamamoto Seiki Kuramitsu Yoshitsugu Shiro Kanagaraj Sekar Shigeyuki Yokoyama 《Acta Crystallographica. Section F, Structural Biology Communications》2007,63(5):446-448
Thermus thermophilus is an aerobic chemoorganotroph that has been found to grow anaerobically in the presence of nitrate. Crystals of glucose dehydrogenase (GDH) from T. thermophilus HB8 belong to space group P21, with unit‐cell parameters a = 36.90, b = 132.96, c = 60.78 Å, β = 97.2°. Preliminary studies and molecular‐replacement calculations reveal that the asymmetric unit contains two monomers. 相似文献
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S. Shailaja S. Venkata Mohan M. Rama Krishna P.N. Sarma 《International biodeterioration & biodegradation》2008,62(2):143-152
Di-ethylhexyl phthalate (DEHP) belongs to the class of phthalate esters and is used as an additive in many products including plastics, paints and inks or as a solvent in industrial formulations. However, it is used mostly for its plasticizing ability in polyvinyl chloride (PVC) products, in which it is often added in concentrations exceeding 40% by mass. DEHP is one of the more recalcitrant phthalate esters, which has xeno-estrogenic, carcinogenic and mutagenic effects. Five different bioslurry reactors were operated under different conditions to study the degradation of DEHP (1 mg g−1 soil) in soil. The process performance was assessed by monitoring DEHP concentration periodically using high performance liquid chromatography (HPLC). The ongoing biological process was monitored by analyzing pH, oxidation–reduction potential (ORP), dissolved oxygen (DO), oxygen uptake rate (OUR) and colony forming units (CFU) for every 24 h. More than 90% degradation was observed within 12 days of the cycle period in the augmented reactors. Metabolites formed during the degradation of DEHP in the slurry phase reactor were identified and the pathway was also established. The degradation process was found to follow zero-order kinetic model. 相似文献
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E. Chabrire A. Volbeda J. C. Fontecilla-Camps M. Roth M.-H. Charon 《Acta Crystallographica. Section D, Structural Biology》1999,55(9):1546-1554
The structure of the homodimeric 267 kDa pyruvate:ferredoxin oxidoreductase (PFOR) of Desulfovibrio africanus was solved with data from two crystals forms, both containing two monomers per asymmetric unit. Phases were obtained from multiwavelength anomalous dispersion (MAD), solvent flattening (SF), molecular replacement (MR) using a 5 Å resolution electron-density search model, multiple isomorphous replacement (MIR) and, finally, electron-density averaging (DA) procedures. It is shown how the combination of all these techniques was used to overcome problems arising from incompleteness of MAD data and weak phasing power of MIR data. A real-space refinement (RSR) procedure is described to improve MR solutions and obtain very accurate protein envelopes and non-crystallographic symmetry (NCS) transformations from 5 Å resolution phase information. These were crucial for the phase extension to high resolution by DA methods. 相似文献