新型光系统II电子传递抑制剂的抑制特性及作用位点的研究

本文由两部分组成，一部分是关于一组新型除草剂(K-15，K-23)的抑制特性及作用位点的研究;另一部分是关于碳酸氢盐对细胞色素b-559高电势的保护作用的研究。 在第一部分，我们首先研究了抑制剂K-23对Psn放氧活性、DCIP光还原和可变荧光等光合特性的影响。研究发现，K-23在低浓度时刺激放氧活性，而在相对高浓度时抑制放氧活性。但是，K-23在低浓度时却有效地抑制了可变荧光。这些数据表明了新型抑制剂的抑制反应是基于氧化还原作用而不是猝灭作用。此外，通过采用胰蛋白酶消化类囊体膜的方式初步检测了新型抑制剂的作用部位，其结果表明：虽然新型抑制剂抑制受体侧电子传递，但它的抑制部位与DCMU不同。 其次，研究了新型抑制剂对光系统II色素蛋白复合体与多肽组分的影响及抑制剂的键合蛋白。应用SDS-PAGE技术，发现新型抑制剂主要作用于光系统II的反应中心蛋白。用温和的Deriphat-PAGE分析也证实了新型抑制剂作用于核心复合物，导致核心复合物二聚体消失。 进一步用SDS-PAGE分析新型抑制剂对Psn多肽组分的影响，发现新型抑制剂主要影响D．、D2、CP43和CP47。用荧光发射的方法确定了K-15键合在D2蛋白上。 最后，结合荧光动力学和HPLC方法，分别从定性和定量的角度，以核心复合物以及抑制剂存在下从BBY中分离的核心复合物为研究对象，详细研究了抑制剂对QA的取代作用。研究发现，在无去垢剂或低浓度去垢剂存在情况下，由于不能创造出适合于QA存在的疏水环境，我们没有得到QA被K-15取代的实验证据。而在抑制剂K-15存在下，用2.2% HTG从BBY分离的核心复合物的实验中，检测不到正的可变荧光Fv，而是得到了降低的FM，这个结果表明QA已被抑制剂在它的作用位点处所取代。 在第二部分，研究了pH5.0—7.0范围内碳酸氢盐对Cyt b-559氧化还原状态转变的影响。首先研究了pH5.0～7.0条件下碳酸氢盐对PSII Cyt b-559还原减氧化差异吸收光谱的影响，发现铁氰化钾还原的PSII Cyt b-559 HP的含量随介质pH值的降低而减少。然而，碳酸氢盐的加入阻止了由于介质pH降低而引起的Cyt b-559由高电势向低电势的转化。比如，当样品温育在pH5.0的介质中，铁氰化钾还原的Cyt b-559 HP含量占总量的25%-30%，当介质中加入2mmol/L碳酸氢盐后Cyt b-559 HP的含量上升，占总量的50%-56%。碳酸氢盐效应对氢醌还原的Cyt b-559HP含量的影响尤为显著。pH6.5时碳酸氢盐对Cyt b-559的还原氧化状态的影响不显著。其次，分别研究了PSII经Tris处理去除锰簇和三个外周蛋白及NH20H处理去除锰簇和17 kDa和23 kDa后，碳酸氢盐对Cytb一559 HP影响的pH依赖值，发现不论在pH5.0还是pH6.5的介质中碳酸氢盐效应都不存在。 综合以上实验结果，我们认为碳酸氢盐对酸化引起的Cyt b-559氧化还原状态的影响与它和锰的作用有关，但也不能排除钙离子与碳酸氢盐之间的协同作用。

Efficient exchange of the primary quinone acceptor QA in isolated reaction centers of Rhodopseudomonas viridis

A key step in the conversion of solar energy into chemical energy by photosynthetic reaction centers (RCs) occurs at the level of the two quinones, QA and QB, where electron transfer couples to proton transfer. A great deal of our understanding of the mechanisms of these coupled reactions relies on the seminal work of Okamura et al. [Okamura, M. Y., Isaacson, R. A., & Feher, G. (1975) Proc. Natl. Acad. Sci. USA 88, 3491–3495], who were able to extract with detergents the firmly bound ubiquinone QA from the RC of Rhodobacter sphaeroides and reconstitute the site with extraneous quinones. Up to now a comparable protocol was lacking for the RC of Rhodopseudomonas viridis despite the fact that its QA site, which contains 2-methyl-3-nonaprenyl-1,4-naphthoquinone (menaquinone-9), has provided the best x-ray structure available. Fourier transform infrared difference spectroscopy, together with the use of isotopically labeled quinones, can probe the interaction of QA with the RC protein. We establish that a simple incubation procedure of isolated RCs of Rp. viridis with an excess of extraneous quinone allows the menaquinone-9 in the QA site to be almost quantitatively replaced either by vitamin K1, a close analogue of menaquinone-9, or by ubiquinone. To our knowledge, this is the first report of quinone exchange in bacterial photosynthesis. The Fourier transform infrared data on the quinone and semiquinone vibrations show a close similarity in the bonding interactions of vitamin K1 with the protein at the QA site of Rp. viridis and Rb. sphaeroides, whereas for ubiquinone these interactions are significantly different. The results are interpreted in terms of slightly inequivalent quinone–protein interactions by comparison with the crystallographic data available for the QA site of the two RCs.

Electron transfer by singlet excited state of porphyn and electron acceptor affinity in rigid medium: photoacoustic phase angle analysis

A espectroscopia fotoacústica obtém informações sobre amplitude e fase, da resposta de um sistema submetido a excitação por luz. Este artigo apresenta estudos do ângulo de fase no processo de transfereência de elétrons entre octaetilporfirina (OEP) e derivados de quinona ambos dispersos em uma matriz polimérica. Observou-se uma tendência no comportamento da fase para valores menores na região espectral próximo de 620 nm. Enquanto que para comprimentos de onda menores este efeito não foi apresentado. Estas medidas sugerem que a transferência de elétrons para o aceitador ocorreu com a participação do estado singleto excitado da octaetilporfirina.

A non-fullerene electron acceptor based on fluorene and diketopyrrolopyrrole building blocks for solution-processable organic solar cells with an impressive open-circuit voltage

A novel solution-processable non-fullerene electron acceptor 6,6′-(5,5′-(9,9-dioctyl-9H-fluorene-2,7-diyl)bis(thiophene-5,2-diyl))bis(2,5-bis(2-ethylhexyl)-3-(thiophen-2-yl)pyrrolo[3,4-c]pyrrole-1,4(2H,5H)-dione) (DPP1) based on fluorene and diketopyrrolopyrrole conjugated moieties was designed, synthesized and fully characterized. DPP1 exhibited excellent solubility and high thermal stability which are essential for easy processing. Upon using DPP1 as an acceptor with the classical electron donor poly(3-hexylthiophene), solution processable bulk-heterojunction solar cells afforded a power conversion efficiency of 1.2% with a high open-circuit voltage (1.1 V). As per our knowledge, this value of open circuit voltage is one of the highest values reported so far for a bulk-heterojunction device using DPP1 as a non-fullerene acceptor.

Crowning of dibenzosilole with a naphthalenediimide functional group to prepare an electron acceptor for organic solar cells

A novel, solution-processable non-fullerene electron acceptor 9,9′-(5,5-dioctyl-5H-dibenzo [b,d]silole-3,7-diyl)bis(2,7-dioctyl-4-(octylamino)benzo[lmn][3,8]phenanthroline-1,3,6,8(2H,7H)-tetraone) (B3) based on dibenzosilole and naphthalenediimide building blocks was designed, synthesized, characterized and successfully used in a bulk-heterojunction organic solar cell. B3 displayed excellent solubility, thermal stability and acquired electron energy levels matching with those of archetypal donor polymer poly(3-hexylthiophene). Solution-processable bulk-heterojunction devices afforded 1.16% power conversion efficiency with a high fill factor of 53%. B3 is the first example in the literature using this design principle, where mild donor units at the peripheries of end-capped naphthalenediimide units tune solubility and optical energy levels simultaneously.

Photoinduced electron transfer in zinc porphyrin-crown ether supramolecule with Eu3+ electron acceptor in crown void

Photoinduced electron transfer (ET) reactions in the zinc porphyrin-crown ether (ZnPCE) supramolecule, in which one crown ether moiety containing Eu3+ as electron acceptor is covalently linked to zinc porphyrin (ZnP), were studied by flash photolysis. In methanol solutions, highly efficient charge separation occurs via intramolecular ET from (ZnP)-Zn-3 to Eu3+ encapsulated in the crown ether void (k(1) = (3 +/- 1) X 10(3) s(-1)) and intramolecular ET from 3ZnP to uncomplexed Eu2+ (k(2) = (2.5 +/- 0.5) X 10(3) s(-1)). Intermolecular ET from Eu2+ escaped from the crown ether void to ZnP.+ (k(tau) = (4.3 +/- 0.7) X 10(8) M(-1) s(-1)) seems to be the main pathway of charge recombination.

Organic photovoltaic cells using hexadecafluorophthalocyaninatocopper (F16CuPc) as electron acceptor material

We fabricated organic photovoltaic cells by using hexadecafluorophthalocyaninatocopper (F16CuPc) as electron acceptor material and para-sexiphenyl (p-6P) as electron donor material. F16CuPc has wide absorption spectrum from 550 nm to 850 nm, which covers the maximum of solar photo flux. The measurement of their external quantum efficiency (EQE) demonstrated that the photocurrent comes from the excitons created in F16CuPc, which were separated into free electrons and holes at heterojunction interface of p-6P and F16CuPc. Moreover, F(16)FuPc with excellent air-stability improved the environmental stability of photovoltaic cells, and the unencapsulated cells exhibited the shelf lifetime of exceeding a week.

Efficient succinic acid production from glycerol by actinobacillus succinogenes using dimethyl sulfoxid as electron acceptor

Dissertação para obtenção do Grau de Mestre em Engenharia Química e Bioquímica

A general synthesis of macrocyclic pi-electron-acceptor systems

Cyclocondensations of aromatic diamines with 1,1'-bis(2,4-dinitrophenyl)-4,4'-bipyridinium salts afford doubly or quadruply charged, macrocyclic, N,N'-diarylbipyridinium cations. These are tolerant of a wide range of acids, bases, and nucleophiles, although they appear to undergo reversible, one-electron reduction by tertiary amines. Single-crystal X-ray analysis demonstrates the presence of a macrocycle conformation in which the 4,4'-bipyridinium and 4,4'-biphenylenedisulfonyl residues are suitably spaced and aligned for complexation with pi-donor arenes, and NMR studies in solution indeed confirm binding to 1,5-bis[hydroxy(ethoxy)ethoxy]naphthalene.

The Regulation of Photosynthetic Electron Transport during Nutrient Deprivation in Chlamydomonas reinhardtii1

The light-saturated rate of photosynthetic O2 evolution in Chlamydomonas reinhardtii declined by approximately 75% on a per-cell basis after 4 d of P starvation or 1 d of S starvation. Quantitation of the partial reactions of photosynthetic electron transport demonstrated that the light-saturated rate of photosystem (PS) I activity was unaffected by P or S limitation, whereas light-saturated PSII activity was reduced by more than 50%. This decline in PSII activity correlated with a decline in both the maximal quantum efficiency of PSII and the accumulation of the secondary quinone electron acceptor of PSII nonreducing centers (PSII centers capable of performing a charge separation but unable to reduce the plastoquinone pool). In addition to a decline in the light-saturated rate of O2 evolution, there was reduced efficiency of excitation energy transfer to the reaction centers of PSII (because of dissipation of absorbed light energy as heat and because of a transition to state 2). These findings establish a common suite of alterations in photosynthetic electron transport that results in decreased linear electron flow when C. reinhardtii is limited for either P or S. It was interesting that the decline in the maximum quantum efficiency of PSII and the accumulation of the secondary quinone electron acceptor of PSII nonreducing centers were regulated specifically during S-limited growth by the SacI gene product, which was previously shown to be critical for the acclimation of C. reinhardtii to S limitation (J.P. Davies, F.H. Yildiz, and A.R. Grossman [1996] EMBO J 15: 2150–2159).

Investigation of electron acceptor states in poly(3-methylthiophene)

Both the DC and AC admittance of Schottky barrier diodes formed at the interface of aluminium and poly(3-methyl thiophene) have been investigated in some detail. The capacitance-voltage plots for the devices suggest the presence of two acceptor states, one shallow and one deep. The total concentration of acceptor states, 10 24-10 26 m -3 depending on the degree of undoping, agrees well with estimates from the reverse I-V characteristics assuming image force lowering of the interfacial potential barrier.

Investigation of electron acceptor states in poly(3-methylthiophene)

Both the DC and AC admittance of Schottky barrier diodes formed at the interface of aluminium and poly(3-methyl thiophene) have been investigated in some detail. The capacitance-voltage plots for the devices suggest the presence of two acceptor states, one shallow and one deep. The total concentration of acceptor states, 10 24-10 26 m -3 depending on the degree of undoping, agrees well with estimates from the reverse I-V characteristics assuming image force lowering of the interfacial potential barrier.

Identification of the proton pathway in bacterial reaction centers: Replacement of Asp-M17 and Asp-L210 with Asn reduces the proton transfer rate in the presence of Cd2+

The reaction center (RC) from Rhodobacter sphaeroides converts light into chemical energy through the reduction and protonation of a bound quinone molecule QB (the secondary quinone electron acceptor). We investigated the proton transfer pathway by measuring the proton-coupled electron transfer, kAB(2) [QA⨪QB⨪ + H+ → QA(QBH)−] in native and mutant RCs in the absence and presence of Cd2+. Previous work has shown that the binding of Cd2+ decreases kAB(2) in native RCs ≈100-fold. The preceding paper shows that bound Cd2+ binds to Asp-H124, His-H126, and His-H128. This region represents the entry point for protons. In this work we investigated the proton transfer pathway connecting the entry point with QB⨪ by searching for mutations that greatly affect kAB(2) (≳10-fold) in the presence of Cd2+, where kAB(2) is limited by the proton transfer rate (kH). Upon mutation of Asp-L210 or Asp-M17 to Asn, kH decreased from ≈60 s−1 to ≈7 s−1, which shows the important role that Asp-L210 and Asp-M17 play in the proton transfer chain. By comparing the rate of proton transfer in the mutants (kH ≈ 7 s−1) with that in native RCs in the absence of Cd2+ (kH ≥ 104 s−1), we conclude that alternate proton transfer pathways, which have been postulated, are at least 103-fold less effective.