金属叶绿素a配位结构的研究

在丙酮溶液中合成得到镧-叶绿素a、钐-叶绿素a和铜-叶绿素a复合物,并研究了它们的紫外可见光谱（UV-Vis）、Fourier红外光谱（FT-IR）和EXAFS结构.镧-叶绿素a、钐-叶绿素a和铜-叶绿素a的UV-Vis谱、FT-IR谱与叶绿素a（含镁）的光谱性质相似,但与脱镁叶绿素a的光谱性质差异很大.证明了La³⁺、Sm³⁺、Cu²⁺已配位到脱镁叶绿素的卟啉环上,形成了镧-叶绿素a、钐-叶绿素a和铜-叶绿素a复合物.通过扩展X射线吸收精细结构谱(EXAFS)研究表明：合成镧-叶绿素a、钐-叶绿素a具有双层夹心结构.La(Ⅲ)、Sm(Ⅲ)夹于两个卟啉环之间, 与上下卟啉环上共八个N原子配位, La-N平均键长0.261 nm,Sm-N平均键长0.243 nm, 而铜-叶绿素a的EXAFS表明为一单层结构,Cu(Ⅱ)与卟啉环中的四个N原子配位,Cu-N平均键长0.197 nm.元素分析也证明镧-叶绿素a、钐-叶绿素a为双层结构,铜-叶绿素a为单层结构.     

石果衣菌藻次级代谢产物的初步研究

荒漠地衣石果衣 Endocarpon pusillum中未能检测出任何次级代谢产物,然而,其共生菌基因组中却含有14个PKS基因和2个NRPS基因。通过激活培养后从中分离出3个次级代谢产物,4,6,9-三羟基-7-甲基-1H,3H-萘并[1,8-cd]吡喃-1,3-二酮（lamellicolic anhydride）、石果衣菌素A（endocarpin A）及石果衣菌素B（endocarpin B）,后二者为新结构化合物。从其共生藻柯氏复球藻 Diplosphaera chodatii中分离出3个脱镁叶绿素类化合物,即脱镁叶绿素甲酯酸b（phaeophorbide b）、13-表-脱镁叶绿素甲酯酸a（13- epi-phaeophorbide a）及脱镁叶绿素甲酯酸a（phaeophorbide a）。其中脱镁叶绿素甲酯酸b具有中等的清除DPPH活性。     

脱镁叶绿酸a单加氧酶与叶绿素降解

近年来叶绿素降解的分子调控机制颇受关注,脱镁叶绿酸a单加氧酶（PaO）作为催化叶绿素卟啉环开环成为四元线性吡咯衍生物的关键酶,成为研究的焦点。本文从PaO的结构与生理功能出发,着重介绍了近期在PaO表达调控方面的最新研究进展,包括PaO突变体中叶绿素降解的反馈机制;去磷酸化作用与PaO的活性调节;非黄化突变体AtNYE1基因、蛋白激酶基因rlpk2、持绿突变体基因sgr的表达与PaO活性之间的关系,并且简要归纳了PaO研究的发展过程与趋势、存在的问题和争议。     

PSⅡ核心复合物能量传递的飞秒时间分辨荧光光谱学研究

运用稳态、瞬态荧光光谱技术对光系统Ⅱ核心复合物的能量传递动力学进行研究.分别用436 nm光脉冲激发叶绿素a分子、45l nm光激发叶绿素a和β-胡萝卜素分子、473和481 nm光激发β-胡萝卜素分子,得到5组反应能量传递、电荷重组等过程的寿命组分:8～40 ps为核心天线中β-胡萝卜素分子通过相邻β-胡萝卜素分子或中间叶绿素a向叶绿素a分子传递能量的时间;85～152 ps为核心天线色素分子激发能传递时间;201～925 ps反映部分电荷重组过程;1.03l～1.2l ns为参与能量传递的色素分子从激发态衰退回到基态的时间;6.17～18.13 ns的长寿命时间组分归因于P680+Pheo-的重组过程.将荧光发射谱进行高斯解析,发现在核心复合物中还至少存在Chla 685 683、Chla 682 680、Chla679 673,677三种特征叶绿素a分子.     

光系统II核心复合物的稳态荧光光谱(英文)

在 83K 和 160K 两个温度下,通过激发波长对荧光发射谱的影响研究了光系统II中核心复合物的荧光光谱特性。用不同波长的光激发,核心复合物的发射谱的最大发射峰值不变,用 480、489、495 和 507nm 的光分别激发核心复合物,其光谱最大峰值处的荧光强度随不同激发波长下β-胡萝卜素分子的吸收强度的增大而降低,在长波长区域光谱的变化依赖于首先被激发的色素分子。所以,激发波长的不同影响着核心复合物中能量传递的途径。通过高斯解析,分析出核心复合物中至少存在有 7组叶绿素a组分,它们是Chla660,Chla670,Chla680,Chla682,Chla684,Chla687和Chla690。     

宽叶吊兰叶绿素生物合成的昼夜节律变化

在被子植物中,从谷氨酰-tRNA到叶绿素的生物合成是由许多酶催化的级联反应,其中间代谢产物具有较强的光反应活性和细胞毒性,因此这一过程在细胞内受到严格的调控。本研究通过检测宽叶吊兰叶片叶绿素生物合成途径的14种中间产物含量随昼夜节律的变化,探讨昼夜节律对宽叶吊兰叶绿素生物合成的影响。结果表明,中间产物ALA(δ-氨基乙酰丙酸)、PBG(胆色素原)、ProtoⅨ(原卟啉Ⅸ)、Heme(血红素)、Mg-ProtoⅨ(镁原卟啉Ⅸ)、Chlide a(叶绿素酸酯a)、Chlide b(叶绿素酸酯b)、Chl a(叶绿素a)、Chl b(叶绿素b)受光诱导,而UrogenⅢ(尿卟啉Ⅲ)、CoprogenⅢ(粪卟啉Ⅲ)和Pchlide(原叶绿素酸脂)受黑暗诱导,尤其是Pchlide在黑暗中的积累量显著增加;Mpe(镁原卟啉Ⅸ单甲酯)和Mpde(镁原卟啉Ⅸ二酯)具有2个积累峰值,分别出现在中午12∶00和夜间24∶00。说明叶绿素生物合成受昼夜节律的调控,但其中间代谢产物含量的变化规律与昼夜节律并不完全一致。     

低浓度NaCl对玉米幼苗光合作用的影响

以Hoagland溶液为基础培养液,采用沙培法,研究了0,5,10,20,30mmol／LNaCl对玉米（Zeamays）幼苗光合作用的影响。结果表明,低浓度NaCl（5,10mmol／LNaCl）处理使其叶绿素含量增加,Chla／Chlb增大,PEPCase活性提高,光合速率增强。说明低盐促进玉米的光合作用可能与叶绿素含量增加,Chla／Chlb增大以及PEPCase活性提高有关。     

PSII核心复合物能量传递的飞秒时间分辩荧光光谱学研究

运用稳态、瞬态荧光光谱技术对光系统Ⅱ核心复合物的能量传递动力学进行研究。分别用436nm光脉冲激发叶绿素a分子、451nm光激发叶绿素a和β-胡萝卜素分子、473和481nm光激发β-胡萝卜素分子,得到5组反应能量传递、电荷重组等过程的寿命组分：8～40 ps为核心天线中β-胡萝卜素分子通过相邻β-胡萝卜素分子或中间叶绿素a向叶绿素a分子传递能量的时间;85～152 ps为核心天线色素分子激发能传递时间;201～925ps反映部分电荷重组过程;1．031～1．21ns为参与能量传递的色素分子从激发态衰退回到基态的时间;6．17～18、13 ns的长寿命时间组分归因于P680^ Pheo^-的重组过程。将荧光发射谱进行高斯解析,发现在核心复合物中还至少存在Chla683^685、Chla680^682、Chla673,677^679三种特征叶绿素a分子。     

东海赤潮高发区沉积物中叶绿素的分析

研究分析了2002年8～9月“赤潮973”航次所采集的沉积物中叶绿素a及其降解产物脱镁叶绿酸含量与分布．结果表明。叶绿素a和脱镁叶绿酸是同源的;随深度增加叶绿素a和脱镁叶绿酸的含量呈递减趋势,到一定深度后含量不再变化。个别站位叶绿素a和脱镁叶绿酸的垂直分布出现了许多小突跃,可能是由生物扰动引起;表层沉积物(0～0．5cm)中的叶绿素a和脱镁叶绿酸的变化幅度分别为0．14～1．17和0．83～5．58μg·g-1。平均值分别为0．54和2．45μg·g-1;并初步探讨水深(光强)、盐度、温度、含水量对叶绿素水平分布的影响;脱镁叶绿酸作为叶绿素的主要降解产物,到一定深度后,成为叶绿素的主要存在形式,约占80％～90％;由于水温的不同,夏季沉积物中的叶绿素a和脱镁叶绿酸含量是春季的3倍之多;比较了上层水柱中的叶绿素和沉积物中的叶绿素的相对含量,平均而言,沉积物中叶绿素占上层水柱中叶绿素的31％．     

基于密度泛函叶绿素a分子活性的量子化学计算

在液相环境下,根据量子化学密度泛函(DFT)的B3LYP泛函在6-31G基组水平的计算结果,从叶绿素a分子的光谱特性、特征基团对前线轨道能量贡献率、激发态、Mulliken电荷分布等量化参数分析比较了其各特征基团的生物活性及吸收波长。结果表明:叶绿素a分子中的5个活性基团中,C(33)、O(34)所组成的酮羰基活性最强,卟啉环的活性次之,C(36)位置上的酯羰基和C(13)位置上的酯羰基及共轭碳碳双键的活性较弱;在发生HOMO→LUMO跃迁后,卟啉环转移的电荷变化总量为0.00012。而酮羰基得到的电荷变化,总量为-0.00062。     

Correlations between functional porphyrin positions and accumulation in cancer cells

Porphyrin is accumulated in tumours due to its interaction with protein. Cancer therapy with porphyrin as a carrier molecule is attracting attention. Porphyrin displays two functional sites termed β- and meso-positions. A correlation between the functional position on the porphyrin molecule and the ability to accumulate in cancer cells is observed in the present study. The accumulation of porphyrin derivatives was determined by measuring fluorescence intensity after incubation for 2 and 24 h. The accumulation of cancer cells depended on the position and length of functional groups. Estimated binding constants between porphyrin and bovine serum albumin suggest that the position of functional groups leads to changes in binding affinity and influences the accumulation of porphyrin derivatives in cancer cells.     

Preparation and study of chiral magnesium porphyrin-amino acid complexes

Mixtures of magnesium protoporphyrin or magnesium mesoporphyrin with a variety of chiral amino acids (L-histidine, D- and L-proline, L-serine and L-threonine) produce prominent induced Cotton effects in the UV-visible region. By contrast magnesium deuteroporphyrin mixtures exhibit no optical rotatory dispersion (ORD)/circular dichroism (CD) spectra. It is proposed that the species producing the Cotton effects are six-coordinate species of the type Mg (porphyrin) (amino acid)₂. For L-histidine and L-threonine CD spectra have shown that complexes of the opposite chirality can be obtained for different samples of magnesium protoporphyrin. For D- and L-proline such a change in sign of spectra was not found for the same magnesium porphyrin samples. Reasons for these observations are presented together with proposals regarding structural details of the six-coordinate complexes. It is also suggested that racemic samples of such amino acid-magnesium porphyrin mixtures could yield optical resolution of products on irradiation with circularly polarised light. Details of a study of this type are presented.     

Studies on the interaction between tetraphenylporphyrin compounds and bovine serum albumin.

The interaction of three porphyrin compounds with bovine serum albumin (BSA) was examined by fluorescence emission spectra at the excitation wavelength 280 nm and in UV-Vis absorption spectra. Through fluorescence quenching experiments, it was confirmed that the combination of three porphyrin compounds with BSA was a single static quenching process. The binding constant K(A), the thermodynamic parameters enthalpy change (DeltaH(0)), Gibbs free energy change (DeltaG(0)) and entropy change (DeltaS(0)) were obtained. It was found that hydrophobic interaction played a main role in tetraphenylporphyrin (TPP) or tetraparacholophenylporphyrin (TClPP) binding to BSA, while tetraparamethoxyphenylporphyrin (TMEOPP) mainly based on van der Waals' force. According to F?ster energy transfer, the separate distance r, the energy transfer efficiency E and F?ster radium R(0) were calculated. The results obtained from the above experiments showed that three porphyrin compounds were tightly bound to BSA.     

福建梅花山57种常绿树叶片叶绿素特征分析

选取福建梅花山海拔1200m和455m地区57种常绿植物为研究对象,测定两海拔的植物叶片叶绿素a和叶绿素b含量,并计算叶绿素a/b、总叶绿素含量。结果表明:(1)植物叶绿素a、b,叶绿素a+b及叶绿素a/b值大多分布在一个相对集中的区域。(2)多数植物新叶叶绿素含量高于老叶,叶绿素a/b值低于老叶,但是新、老叶无显著差异。(3)不同海拔的植物叶片总叶绿素含量和叶绿素a/b值有明显差异,低海拔地区明显高于高海拔地区。植物通过总叶绿素含量和a/b值的变化以适应不同环境条件。     

不同盐度下水华束丝藻对CO2浓度倍增的生理响应

本文研究了在盐度分别为2‰和4‰的条件下,CO2浓度倍增对水华束丝藻(Aphanizomenon flos-aquae)生长速率、光合活性及光合色素比例与丙二醛含量的影响。结果表明随着盐度的增加水华束丝藻的生长速率和光合活性受到显著抑制,叶绿素a与藻蓝素比例大幅降低,丙二醛含量明显提高。在实验盐度范围内,CO2浓度倍增显著促进水华束丝藻的生长速率和最大光合电子传递速率(ETRmax)与色素比例,而且盐度越高促进效果越明显。此外CO2浓度倍增能显著降低丙二醛(MDA)含量。从而减少膜脂过氧化,缓解盐度胁迫。     

Site-directed photochemical coupling of cytochrome b6f-associated chlorophyll

Cytochrome b(6)f complexes contain a molecule of chlorophyll a (Chla), which, in Chlamydomonas reinhardtii, can be exchanged for extraneous chlorophyll during protracted incubation of the purified complex in detergent solution. The specificity of the site and its location in the complex have been studied by photochemical coupling and circular dichroism spectroscopy. Following substitution of the original chlorophyll with [(3)H]Chla, the complex was irradiated in the Soret absorption band of Chla to complete bleaching and the amount of radioactivity covalently bound to each b(6)f subunit determined. Strong labeling was found to be associated with cytochrome f. The labeling originates from [(3)H]Chla molecules bound to a slowly exchanging site and showing the properties of the endogenous Chl, not from molecules dissolved in the detergent belt surrounding the complex. Chlorophyll b (Chlb) can compete with Chla, albeit with a lower affinity. Irradiation of [(3)H]Chlb introduced into the slowly exchanging site yielded the same labeling pattern that was observed with [(3)H]Chla. Proteolytic cleavage showed [(3)H]Chla labeling to be strictly restricted to the C-terminal region of cytochrome f. Circular dichroism spectra of the native complex revealed a bilobed signal characteristic of excitonic interaction between chlorophylls. The structural and evolutionary implications of these findings are discussed.     

Energy Transfer among Chlorophylls in Trimeric Light-harvesting ComplexⅡ of Bryopsis corticulans

A study on energy transfer among chlorophylls(Chls)in the trimeric unit of the major light-harvesting complex Ⅱ(LHC Ⅱ)from Bryopsis corriculan,was carried out using time-correlated singlephoton counting.In the chlorophyll Q region of LHC Ⅱ,six molecules characterized as Chlb_(628),Chlb_(646),Chlb_(652)~(654,657),Chla_(664)~(666),Chla_(674)~(677.680)and Chla_(682)~(683) were discriminated according to their absorption spectrumand fluorescence emission spectrum.Then,excited by pulsed light of 628 nm,fluorescence kinetics spectrain the chlorophyll Q region were measured.In accordance with the principles of fluorescence kinetics,thesekinetics data were analyzed with a multi-exponential model.Time constants on energy transfer were obtained.An overwhelming percentage of energy transfer among chlorophylls undergoes a process longer than 97picoseconds(ps),which shows that,before transferring energy to another Chl,the excited Chl might convertenergy to vibrations of a lower state with different multiplicity(intersystem crossing).Energy transfer at thelevel of approximately 10 ps was also obtained,which was interpreted as the excited Chls may go throughinternal conversion before transferring energy to another Chl.Although with a higher standard deviation,timeconstants at the femtosecond level can not be entirely excluded,which can be attributed to the ultrafastprocess of direct energy transfer.Owing to the arrangement and direction of the dipole moment of Chls inLHC Ⅱ,the probability of these processes is different.The fluorescence lifetimes of Chlb_(652)~(654,657),Chla_(664)~(666),Chla_(674)~(677.680)and Chla_(682)~(683)were determined to be 1.44ns,1.43 ns,636 ps and 713 ps,respectively.Thepercentages of energy dissipation in the pathway of fluorescence emission were no more than 40% in thetrimeric unit of LHC Ⅱ.These results are important for a better understanding of the relationship between thestructure and function of LHC Ⅱ.     

A crystallographic study of deoxy cobalt (II) mesoporphyrin IX myoglobin.

The crystal structures of acid metmyoglobin and deoxy cobalt(II)mesoporphyrin IX myoglobin were compared by a difference Fourier analysis at 2.5 A resolution. No large differences in protein conformation were observed. The greatest density of structural differences was found in the heme region. There was a loss of the histidine-bound sulfate ion and of the metal-bound water molecule, as well as a shift in the position of the prosthetic group with associated changes in the adjacent globin. The structural changes resulting from the substitution of ethyl for the vinyl side chains of the porphyrin were clearly observed. There was also a suggestion of a conformational change of the porphyrin ring. It was not clear whether there was any change of the metal position relative to the porphyrin plane or proximal histidine.     

The enzymatic one-electron reduction of porphyrins to their anion free radicals

The anaerobic enzymatic one-electron reduction of uroporphyrin I (in the absence of light) by the ferredoxin/ferredoxin:NADP+ oxidoreductase system was investigated using NADPH as the source of reducing equivalents. The porphyrin anion free radical metabolite formed by one-electron reduction of the parent molecule was detected with ESR spectroscopy. The ESR spectrum exhibited a singlet (g = 2.0021) with a 5.4-G peak-to-peak linewidth. The reduction process was also investigated under aerobic conditions. The reduction of molecular oxygen to superoxide anion radical by the porphyrin anion radical was demonstrated by using the ESR technique of spin trapping. The ESR spectra of the spin-trapped oxygen-derived radicals were superoxide dismutase-sensitive and catalase-insensitive, supporting the assignment of the trapped radical to the superoxide anion radical. These aerobic experiments demonstrate electron transfer from the porphyrin anion radical to molecular oxygen. The anaerobic reduction of Photofrin II by hepatic microsomes and the ferredoxin/ferredoxin:NADP+ oxidoreductase system to a porphyrin anion radical was also investigated. Free radical formation by ferredoxin: NADP+ oxidoreductase is totally dependent upon ferredoxin. The ESR spectrum of this porphyrin free radical also exhibited a singlet (g = 2.0026) with a 15-G peak-to-peak linewidth.     

Modification of Photosystem I Light Harvesting of Bundle-Sheath Chloroplasts Occurred during the Evolution of NADP-Malic Enzyme C4 Photosynthesis

Low-temperature emission spectra and excitation spectra for chlorophyll fluorescence were recorded from leaves of species of the genus Flaveria (Asteraceae) with C3, C3-C4-intermediate, C4-like, and C4 photosynthesis. Among the latter two groups, high chlorophyll b absorption was observed in excitation spectra for photosystem I (PSI) fluorescence. By comparing leaf data with those from isolated chloroplast fractions, the high chlorophyll b absorption was attributed to the specific properties of the bundle-sheath chloroplasts in leaves from C4 plants. The deconvolution of the PSI excitation spectra and the use of a model revealed that the contribution of photosystem II absorption to the functional antenna of PSI was markedly increased in leaves from three of the five C4-like and C4 species investigated in detail. The two other species exhibited normal, C3-like light-harvesting properties of PSI. The former species are known for efficient carbon assimilation, the latter for decreased efficiencies of carbon assimilation. It is concluded that photosystem II becomes a substantial part of the functional PSI antenna late in the evolution of C4 photosynthesis, and that the composite antenna optimizes the light-harvesting of PSI in bundle-sheath chloroplasts to meet the energy requirements of C4 photosynthesis.