The structure and function of the cytochrome <Emphasis Type="Italic">c</Emphasis><Subscript>2</Subscript>: reaction center electron transfer complex from <Emphasis Type="Italic">Rhodobacter sphaeroides</Emphasis>

In the photosynthetic bacterium, Rhodobacter sphaeroides, the mobile electron carrier, cytochrome c₂ (cyt c₂) transfers an electron from reduced heme to the photooxidized bacteriochlorophyll dimer in the membrane bound reaction center (RC) as part of the light induced cyclic electron transfer chain. A complex between these two proteins that is active in electron transfer has been crystallized and its structure determined by X-ray diffraction. The structure of the cyt:RC complex shows the cyt c₂ (cyt c₂) positioned at the center of the periplasmic surface of the RC. The exposed heme edge from cyt c₂ is in close tunneling contact with the electron acceptor through an intervening bridging residue, Tyr L162 located on the RC surface directly above the bacteriochlorophyll dimer. The binding interface between the two proteins can be divided into two regions: a short-range interaction domain and a long-range interaction domain. The short-range domain includes residues immediately surrounding the tunneling contact region around the heme and Tyr L162 that display close intermolecular contacts optimized for electron transfer. These include a small number of hydrophobic interactions, hydrogen bonds and a pi-cation interaction. The long-range interaction domain consists of solvated complementary charged residues; positively charged residues from the cyt and negatively charged residues from the RC that provide long range electrostatic interactions that can steer the two proteins into position for rapid association.     

Extracellular Fe(III) reductase structure reveals a modular organization enabling S-layer insertion and electron transfer to insoluble substrates

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微生物种间电子传递的研究进展

种间电子传递可促进微生物发生共代谢,因而在地球生物化学循环和环境污染修复中具有重要意义。根据电子传递方式的不同可将种间电子传递分为直接种间电子传递（direct interspecies electron transfer,DIET）和间接种间电子传递（mediated interspecies electron transfer,MIET）,其中,直接种间电子传递由于易发生、效率高而受到更加广泛的关注。本文总结了近年来关于种间电子传递的研究进展,阐述了种间电子传递的途径,比较了DIET和MIET的优缺点,并对开发更多具有种间电子传递功能的微生物提出了建议,以期加深人们对于种间电子传递的理解,并对未来该领域的研究提供参考。     

微生物互营产甲烷过程中的种间电子传递

甲烷作为全球第二大温室气体,是典型的可再生清洁能源,也是碳循环中的重要物质组成。大气中约74%的甲烷由产甲烷古菌和其他微生物的互营产生,种间电子传递(interspecies electron transfer, IET)是微生物菌群降低热力学能垒、实现互营产甲烷的核心过程。IET可分为间接种间电子传递(mediated interspecies electron transfer,MIET)和直接种间电子传递(direct interspecies electron transfer, DIET)两种类型,其中MIET依赖氢气、甲酸等载体完成电子的远距离传输,而DIET则依赖导电菌毛、细胞色素c等膜蛋白,通过微生物的直接接触实现电子传递。本文将从IET的研究历程出发,从电子传递机制、微生物种类、生态多样性等方面对微生物互营产甲烷过程中的两种IET类型进行比较,最后对未来待探索的方向进行展望。本综述有助于加深对微生物互营产甲烷过程中IET的理解,为解决由甲烷引发的全球气候变暖等生态问题提供理论支撑。     

Extracellular electron transfer of Methylophilus methylotrophs

Methylophilus methylotrophs, as representatives of C1 microbes, widely exist in the interface of hyperoxia–hypoxia environment and plays a key role in global carbon cycle. Most studies of Methylophilus were carried out under hyperoxic conditions, while it remains unclear how Methylophilus survive under oxygen-limited conditions. In this study, we report that ferrihydrite could serve as an alternative electron acceptor for Methylophilus strain with reduction rate of 53.6 μM_Fe(II) d⁻¹, when cells were deprived of oxygen. Genomic analysis and electrochemical measurements suggested that two strategies, cytochrome c and riboflavin, were involved in the extracellular mineral respiration. Respiration of extracellular electron acceptor by a Methylophilus strain demonstrated in this study broadens our understanding of energy metabolism by Methylophilus and suggests as a survival strategy for these microbes under oxygen-limited conditions. The results also demonstrate Methylophilus spp. as novel type of electrochemical active microbes that could be applied in bioelectrochemical systems with single-carbon substrate.     

Control of electron transfer in metalloproteins

The role of protein structure in the control of electron transfer in metalloproteins is briefly discussed, with reference to existing theoretical models and available three dimensional information.     

微生物种间直接电子传递研究进展

一直以来氢气和甲酸被认为是微生物间电子传递的中间电子传递体.近年来的研究发现,微生物之间可以通过种间直接电子传递(DIET)来替代氢气/甲酸传递.DIET作为一种新发现的微生物间电子传递途径,其电子传递效率要高于传统的种间氢气/甲酸传递.DIET这一新发现改变了微生物互营生长代谢必须依赖氢气或甲酸等电子载体的传统认识,...     

Crystallization and preliminary X-ray analysis of electron transfer flavoproteins from human and Paracoccus denitrificans.

Mammalian electron transfer flavoprotein (ETF) is a soluble, heterodimeric flavoprotein responsible for the oxidation of at least nine primary matrix flavoprotein dehydrogenases. Crystals have been obtained for the recombinant human electron transfer flavoprotein (ETFhum) by the sitting-drop vapor diffusion technique using polyethylene glycol (PEG) 1500 at pH 7.0 as the precipitating agent. ETFhum crystallizes in the monoclinic space group P2(1), with unit cell parameters a = 47.46 angstrum, b = 104.10 angstrum, c = 63.79 angstrum, and beta = 110.02 degrees. Based on the assumption of one alpha beta dimer per asymmetric unit, the Vm value is 2.69 angstrum 3/Da. A native data set has been collected to 2.1 angstrum resolution. One heavy-atom derivative has also been obtained by soaking a preformed crystal of ETFhum in 2 mM thimerosal solution for 2h at 19 degrees C. Patterson analysis indicates one major site. The analogous electron transfer flavoprotein from Paracoccus denitrificans (ETFpar) has also been crystallized using PEG 8000 at pH 5.5 as the precipitating agent. ETFpar crystallizes in the orthorhombic space group P2(1)2(1)2(1), with unit cell parameters a = 79.98 angstrum, b = 182.90 angstrum, and c = 70.07 angstrum. The Vm value of 2.33 angstrum 3/Da is consistent with two alpha beta dimers per asymmetric unit. A native data set has been collected to 2.5 angstrum resolution.     

细菌胞际电子转移及其生态生理学意义研究进展

胞际电子转移是指细胞内电子以间接或直接的方式传递到细胞外,最终到达细胞周围电子受体的过程.胞际电子转移普遍存在于自然界,尤其存在于电子受体相对匮乏的环境中.胞际电子转移可分为间接和直接胞际电子转移.间接胞际电子转移(胞际基质转移)是主要借助氢、甲酸以及其他代谢产物的电子传递;而直接胞际电子转移则由胞内电子转移偶联胞外电子传递实现.胞际电子转移促进了细胞的基质代谢活性,拓展了细胞的作用空间,具有重要的生理意义.胞际电子转移产生了电流,实现了菌间能源共享,驱动了胞外物质(如重金属、腐殖质)转化,具体重大的生态意义.本文总结相关文献,对细菌胞际电子转移的过程、特点、机理及其生态生理学意义作了系统的分析和探讨.     

Molecular mechanism of intramolecular electron transfer in dimeric sulfite oxidase

Sulfite oxidase (SOX) is a homodimeric molybdoheme enzyme that oxidizes sulfite to sulfate at the molybdenum center. Following substrate oxidation, molybdenum is reduced and subsequently regenerated by two sequential electron transfers (ETs) via heme to cytochrome c. SOX harbors both metals in spatially separated domains within each subunit, suggesting that domain movement is necessary to allow intramolecular ET. To address whether one subunit in a SOX dimer is sufficient for catalysis, we produced heterodimeric SOX variants with abolished sulfite oxidation by replacing the molybdenum-coordinating and essential cysteine in the active site. To further elucidate whether electrons can bifurcate between subunits, we truncated one or both subunits by deleting the heme domain. We generated three SOX heterodimers: (i) SOX/Mo with two active molybdenum centers but one deleted heme domain, (ii) SOX/Mo_C264S with one unmodified and one inactive subunit, and (iii) SOX_C264S/Mo harboring a functional molybdenum center on one subunit and a heme domain on the other subunit. Steady-state kinetics showed 50% SOX activity for the SOX/Mo and SOX/Mo_C264S heterodimers, whereas SOX_C264S/Mo activity was reduced by two orders of magnitude. Rapid reaction kinetics monitoring revealed comparable ET rates in SOX/Mo, SOX/Mo_C264S, and SOX/SOX, whereas in SOX_C264S/Mo, ET was strongly compromised. We also combined a functional SOX Mo domain with an inactive full-length SOX R217W variant and demonstrated interdimer ET that resembled SOX_C264S/Mo activity. Collectively, our results indicate that one functional subunit in SOX is sufficient for catalysis and that electrons derived from either Mo^(IV) or Mo^(V) follow this path.     

Regulation of electron transfer by the quinone pool

Strong evidence for a random collisional mechanism for ubiquinone-mediated electron transfer is provided by the characteristic kinetic properties of respiratory chains originally explored by Kröger, A., and Klingenberg, M. (1973),Eur. J. Biochem. 34, 313–323. A kinetic model which leads to this so-called simple Q-pool behavior has been described and we use this in reviewing evidence that electron transfer is diffusion-controlled as well as diffusion-coupled. We also consider mechanisms by which the kinetics of electron transfer might deviate from simple Q-pool behavior and how these might be implicated in the regulation of electron transport.     

微生物种间直接电子传递机理及应用研究进展

微生物胞内产生的电子转移到其他电子受体而获得能量的过程称为微生物胞外电子传递,其中,另一微生物作为电子受体时发生的电子传递称为微生物种间电子传递。根据微生物种间电子传递机制,可分间接种间电子传递和种间直接电子传递。由于种间直接电子传递不需要其他物质介导,因此较间接种间电子传递效率更高、能量利用更高。本文系统阐述了微生物进行胞外电子传递的机理及应用,重点分析了种间直接电子传递机理,并概述种间直接电子传递应用领域,为寻找更多电连接的微生物群落以及应用微生物提供参考。     

Conformational reorganisation in interfacial protein electron transfer

Protein-protein electron transfer (ET) plays an essential role in all redox chains. Earlier studies which used cross-linking and increased solution viscosity indicated that the rate of many ET reactions is limited (i.e., gated) by conformational reorientations at the surface interface. These results are later supported by structural studies using NMR and molecular modelling. New insights into conformational gating have also come from electrochemical experiments in which proteins are noncovalently adsorbed on the electrode surface. These systems have the advantage that it is relatively easy to vary systematically the driving force and electronic coupling. In this review we summarize the current knowledge obtained from these electrochemical experiments and compare it with some of the results obtained for protein-protein ET.     

矿物电子能量协同微生物胞外电子传递与生长代谢

矿物是无机自然界吸收与转化能量的重要载体,其与微生物的胞外电子传递过程体现出矿物电子能量对微生物生长代谢与能量获取方式的影响。根据电子来源与产生途径,以往研究表明矿物中变价元素原子最外层或次外层价电子与半导体矿物导带上的光电子是微生物可以利用的两种不同胞外电子能量形式,其产生及传递方式与微生物胞外电子传递的电子载体密切相关。在协同微生物胞外电子传递过程中,矿物不同电子能量形式之间既有相似性亦存在着差异。反过来,微生物胞内-胞外电子传递途径也影响对矿物电子能量的吸收与获取,进而对微生物生长代谢等生命活动产生影响。本文在阐述矿物不同电子能量形式产生机制及其参与生物化学反应的共性和差异性特征基础上,综述了微生物获取矿物电子能量所需的不同电子载体类型与传递途径,探讨了矿物不同电子能量形式对微生物生长代谢等生命活动的影响,展望了自然条件下微生物利用矿物电子能量调节其生命活动、调控元素与能量循环的新方式。     

Potential-dependent extracellular electron transfer pathways of exoelectrogens

Exoelectrogens are distinct from other bacteria owing to their unique extracellular electron transfer (EET) abilities that allow for anaerobic respiration with various external redox-active surfaces, including electrode and metal oxides. Although the EET process is known to trigger diverse extracellular redox reactions, the reverse impact has been long overlooked. Recent evidences show that exoelectrogens can sense the potential changes of external surfaces and alter their EET strategies accordingly, which imparts them remarkable abilities in adapting to diverse and redox-variable environment. This mini-review provides a condensed overview and critical analysis about the recent discoveries on redox-dependent EET pathways of exoelectrogens, with focus on Geobacter sulfurreducens and Shewanella oneidensis. We summarize the detailed responses of various EET components, analyze the drives and mechanisms of such responses, highlight the diversity of EET dynamics among different bacterial species and under integrated effects of redox potential and surface chemistry, and discusses the future research needs.     

PQQ glucose dehydrogenase with novel electron transfer ability

PQQ glucose dehydrogenase from Acinetobacter calcoaceticus (GDH-B) is one of the most industrially attractive enzymes, as a sensor constituent for glucose sensing, because of its high catalytic activity and insensitivity to oxygen. We attempted to engineer GDH-B to enable electron transfer to the electrode in the absence of artificial electron mediator by mimicking the domain structure of the quinohemoprotein ethanol dehydrogenase (QH-EDH) from Comamonas testosteroni, which is composed of a PQQ-containing catalytic domain and a cytochrome c domain. We genetically fused the cytochrome c domain of QH-EDH to the C-terminal of GDH-B. The constructed fusion protein showed not only intra-molecular electron transfer, between PQQ and heme of the cytochrome c domain, but also electron transfer from heme to the electrode, thereby allowing the construction of a direct electron transfer-type glucose sensor.     

Mechanism of photoinduced electron transfer in photosystem II reaction centers

The shape of the EPR spectrum of the triplet state of photosystem II reaction centers with a singly reduced primary acceptor complex Q_AFe²⁺ was studied. It was shown that the spectroscopic properties do not significantly change when the relaxation of the primary acceptor is accelerated and when the magnetic interaction between the reduced quinone molecule Q_A and the nonheme iron ion Fe²⁺ is disrupted. This observation confirmed the earlier conclusion that the anisotropy of the quantum yield of the triplet state is the main cause of the anomalous shape of the EPR spectrum. A scheme of primary processes in photosystem II that is consistent with the observed properties of the EPR spectrum of the triplet state is discussed.     

Electroactive biofilms: how microbial electron transfer enables bioelectrochemical applications

Microbial biofilms are ubiquitous. In marine and freshwater ecosystems, microbe–mineral interactions sustain biogeochemical cycles, while biofilms found on plants and animals can range from pathogens to commensals. Moreover, biofouling and biocorrosion represent significant challenges to industry. Bioprocessing is an opportunity to take advantage of biofilms and harness their utility as a chassis for biocommodity production. Electrochemical bioreactors have numerous potential applications, including wastewater treatment and commodity production. The literature examining these applications has demonstrated that the cell–surface interface is vital to facilitating these processes. Therefore, it is necessary to understand the state of knowledge regarding biofilms’ role in bioprocessing. This mini-review discusses bacterial biofilm formation, cell–surface redox interactions, and the role of microbial electron transfer in bioprocesses. It also highlights some current goals and challenges with respect to microbe-mediated bioprocessing and future perspectives.     

A structural basis of light energy and electron transfer in biology

Aspects of intramolecular light energy and electron transfer will be discussed for three protein cofactor complexes, whose three-dimensional structures have been elucidated by x-ray crystallography: Components of light harvesting cyanobacterial phycobilisomes, the purple bacterial reaction centre, and the blue multi-copper oxidases. A wealth of functional data is available for these systems which allow specific correlations between structure and function and general conclusions about light energy and electron transfer in biological materials to be made.     

Effects of protein-protein interactions on electron transfer: docking and electron transfer calculations for complexes between flavodoxin and c-type cytochromes

 Theoretical studies of protein-protein association and electron transfer were performed on the binary systems formed by Desulfovibrio vulgaris Hildenborough (D. v. H.) flavodoxin and D. v. H. cytochrome c ₅₅₃ and by flavodoxin and horse heart cytochrome c. Initial structures for the complexes were obtained by rigid-body docking and were refined by MD to allow for molecular flexibility. The structures thus obtained were analysed in terms of their relative stability through the calculation of excess energies. Electrostatic, van der Waals and solvation energy terms showed all to have significant contributions to the stability of complexes. In the best association solutions found for both cytochromes, these bind to different zones of flavodoxin. The binding site of flavodoxin observed for cytochrome c is in accordance with earlier works [27]. The various association modes found were characterised in terms of electron transfer using the Pathways model. For complexes between flavodoxin and horse heart cytochrome c, some correlation was observed between electron tunnelling coupling factors and conformation energy; the best conformation found for electron transfer corresponded also to the best one in terms of energy. For complexes between flavodoxin and cytochrome c ₅₅₃ this was not the case and a lower correlation was observed between electron tunnelling coupling factors and excess energies. These results are in accordance with the differences in the experimental dependence of electron transfer rates with ionic strength observed between these two cases. Received: 29 December 1998 / Accepted: 22 March 1999