菊科植物的杀菌活性及其活性成分

本文对菊科16个属———刺苞果属(Acanthospermum)、蓍属(Achillea)、胜红蓟属(Ageratum)、蒿属(Arte-misia)、紫苑属(Aster)、Balsamorhiza、雏菊属(Bellis)、鬼针草属(Bidens)、艾纳香属(Blumea)、天明精属(Carpe-sium)、红花属(Carthamus)、矢车菊属(Centaurea)、茼蒿属(Chrysanthemum)、菊苣属(Cichorium)、秋英属(Cos-mos)、大丽花属(Dahlia)中具有杀菌活性的植物按属的顺序进行了综述.表明在16个属中主要杀菌植物集中在蓍属(Achillea)、胜红蓟属(Ageratum)、蒿属(Artemisia)、天明精属(Carpesium)和矢车菊属(Centaurea)等5个属中,为进一步开展菊科植物资源的植物性杀菌剂研究和开发利用提供参考.     

鼠尾草酸的抗氧化活性及抑菌活性研究

通过自由基清除能力、还原力和平板二倍稀释法实验,评价了鼠尾草酸的抗氧化活性、抑菌活性。结果显示:相同浓度的鼠尾草酸抗氧化活性明显高于合成抗氧化剂PG、BHT、BHA、VE,略低于THBQ,其清除DPPH的IC50值为2.53μg/m L,清除ABTS的IC50值为51.58μg/m L。抑菌实验表明:鼠尾草酸具有广谱抗菌性,特别是大肠埃希氏杆菌、绿脓假单胞菌,其最低抑菌浓度分别为4、2μg/m L,表现出了比新洁尔灭、氨苄青霉素钠、红霉素更好的抑菌活性;对于肺炎克雷伯氏菌,则表现出了与红霉素同样的抑菌效果,优于新洁尔灭和氨苄青霉素钠。     

蝉蜕活性成分的提取及其抑菌活性的研究

本文报道了对传统中药蝉蜕抑菌活性成分提取与抑菌活性研究结果。对4种提取方法和7种提取溶剂的提取效果进行了平行比较研究,研究结果显示,以体积分数95%乙醇作为提取溶剂,4种提取方法中,以超声波法提取时的浸膏得率最高。采用超声波提取法时,在7种提取溶剂中,以95%乙醇提取浸膏的得率最高。抑菌试验结果表明,不同方法获得提取物与对照组相比对Escheridia coli均有明显的抑菌作用(P≤0.05),但不同提取方法得到的提取物之间,并无显著的抑菌活性差异。其中采用超声波提取法,以95%乙醇为提取溶剂时,所得提取物的抑菌活性最高,对E.coli的最小抑菌浓度为0.078 mg/mL。该研究结果是蝉蜕抑菌成分的首次报道。     

牛乳铁蛋白素抗菌活性及抗菌活性位点

牛乳铁蛋白素(Bovine Lactoferrcin, LfcinB)是乳铁蛋白在酸性环境下经胃蛋白酶作用N端释放的一段多肽, 它具有多种生物学功能。研究LfcinB广谱抗菌性及改变LfcinB氨基酸序列对其抗菌能力的影响, 寻找LfcinB抗菌作用的结构位点。人工合成LfcinB, 采用琼脂扩散法测定LfcinB抗菌图谱。人工合成丙氨酸取代3位半胱氨酸的LfcinB、丙氨酸取代8位色氨酸的LfcinB和去掉2个半胱氨酸的LfcinB样品, 测定最小抑菌浓度, 确定LfcinB抗菌活性位点。研究结果表明:     

凝集素碎片的糖结合活性

用固相合成法分别合成了羊蹄甲、小扁豆和欧洲百脉根３种植物凝集素中的某些糖结合活性部位的肽段。用毛细管电泳法观察到这些肽段和拟糖蛋白以及寡糖之间有一定的结合能力,而且表现出相对的专一性。     

海洋细菌活性蛋白、活性肽研究的若干新进展

海洋蕴藏着丰富的生物活性资源,近年来海洋细菌活性蛋白及活性肽的研究在资源开发与利用领域中很受关注。本文首先分析了海洋细菌来源的各种活性蛋白及活性肽在不同层面上的生物学活性特征,其次对海洋活性蛋白及活性肽的获得方式及其在医药学、工业、水产养殖业及环境保护等领域中的应用现状与潜力作简要介绍,最后对该领域研究前景进行了展望。     

虫草的抗菌抗肿瘤活性研究进展

虫草代谢产物丰富,很多具有抗菌和（或）抗肿瘤活性,有关其生物活性物质的研究受到广泛关注。对有抗菌和（或）抗肿瘤活性研究报道的虫草作了系统总结。     

苦瓜降血糖活性评述

本文对近年有关苦瓜降血糖活性,活性物及其作用机理,毒副作用等研究成果作了综述。在文献分析的基础上本文提出,苦瓜蛋白(或多肽)口服降糖活性的作用机理值得深入探讨,苦瓜皂苷的药理活性则应当用纯品深入研究,而寻找能够提高糖尿病动物血浆胰岛素水平的活性物,仍然是苦瓜研究中值得注意的课题。此外,对苦瓜的毒副作用人们应当引起足够的重视。     

海洋浮游藻类细胞质膜氧化还原活性与细胞外CA活性的关系

海洋浮游藻类除通过吸收和释放分子与离子来改变其环境的化学成分外,还可通过细胞外表面一些酶的作用引起质膜外化学物质变化。在这方面,海洋浮游藻类一个主要的细胞外表面酶-碳酸酐酶(CA),在经胰蛋白酶处理从细胞质膜上释放出来后,仍保留其催化活性。当细胞外表面CA(简称细胞外CA)具活性时,可催化质膜外HCO_3~-与CO_2的相互转化,为Rubisco(磷酸核酮糖羧化酶)提供一稳定的CO_2流量环境,以维持正常的光合作用。     

INORGANIC CARBON LIMITATION, EXOFACIAL CARBONIC ANHYDRASE ACTIVITY, AND PLASMA MEMBRANE REDOX ACTIVITY IN MARINE PHYTOPLANKTON SPECIES

In the marine phytoplankton species tested, a possible link between exofacial ferricyanide reduction at the plasma membrane of intact cells, inorganic carbon status of the cells, and extracellular carbonic anhydrase (CA) activity is proposed. In species with no extracellular CA activity under carbon-limited or carbon-replete conditions, barely detectable ferricyanide reduction was observed. Species in which extracellular CA was only detected under carbon-limited conditions showed high rates of exofacial ferricyanide reduction, as shown previously for Skeletonema costatum. Immunological analysis has demonstrated that the CA protein was present in both carbon-limited and carbon-replete cells, even though the CA activity could only be detected when inorganic carbon was limiting. Incubation of the inactive extracellular CA protein from carbon-replete cells with DTT caused activation of the enzyme. It is proposed that CA limitation in the light promotes proton extrusion and increased plasma membrane redox activity, which result in the protonation and activation of the extracellular CA.     

BICARBONATE UTILIZATION BY MARINE PHYTOPLANKTON SPECIES

The contribution of bicarbonate to total dissolved inorganic carbon (DIC) utilization was investigated using 18 marine phytoplankton species, including members of Bacillariophyceae, Dinophyceae, Prymnesiophyceae, and Raphidophyceae, under carbon-replete or -limited conditions. Extracellular carbonic anhydrase (CA) was assayed as an indicator of extracellular CA-catalyzed HCO⁻₃ utilization. For some species, extracellular CA was constitutive, in others activity was detected under conditions of carbon limitation, and in others, even under carbon-limited conditions, activity was not detected. In species without extracellular CA, direct HCO⁻₃ uptake was investigated using a pH drift technique in a closed system, DIC measurements, and the use of the anion exchange inhibitor 4'4'-diisothiocyanatostilbene-2,2-disulfonic acid (DLDS). Three of these species (Chaetoceros compressus, Thalassiosira pseudonana, and Glenodinium foliaceum) gave a pH drift not inhibited by DIDS, but cultures of Chrysochromulina kappa, Gephrocapsa oceanica, and Coccolithus pelagicus, in which DLDS inhibited DIC uptake, did not give a pH drift. This result shows that direct HCO₃⁻ transport may occur by an anion exchange-type mechanism in some species but not others. Of the eighteen species investigated, only Heterosigma akashiwo did not have the potential for direct uptake or extracellular CA-catalyzed HCO⁻₃ utilization.     

PHYTOPLANKTON PLASMA MEMBRANE REDOX ACTIVITY: EFFECT OF IRON LIMITATION AND INTERACTION WITH PHOTOSYNTHESIS1

Phytoplankton plasma membrane electron transport activity was determined by monitoring the reduction of the impermeant artificial electron acceptor ferricyanide in a range of diatoms. The results revealed that constitutive plasma membrane electron transport activity of marine diatoms is high compared with chlorophytes and higher plant cells. Diatom plasma membrane electron transport activity was not significantly increased by iron limitation. This lack of induction on iron limitation indicates that diatoms have an iron acquisition strategy that is distinct from chlorophytes and the dicotyledon higher plants that exhibit marked increases in plasma membrane ferricyanide reductase activity on iron limitation. The interaction of the constitutive plasma membrane electron transport with photosynthesis was also investigated. We found that 1) ferricyanide reduction at the plasma membrane was progressively inhibited in response to increasing irradiances; 2) the presence of extracellular ferricyanide, but not the reduced couple ferrocyanide, caused a marked inhibition of carbon fixation at high irradiance; and 3) extracellular electron acceptors ferricyanide and hexachloroiridate (but not ferrocyanide) induced an immediate and reversible decrease in fluorescence yields (F_o and F_m). The extent to which extracellular electron acceptors affected CO₂ fixation, F_o, and F_m was related to the level of constitutive ferricyanide reductase activity, the species with highest ferricyanide reduction rates being most sensitive. The data suggest that consumption of electrons and/or reductant at the plasma membrane by external acceptors may compete directly with CO₂ fixation for electrons, alter cytosolic‐chloroplast redox poise, and/or induce a redox‐signaling cascade that alters photosynthetic metabolism.     

海洋浮游藻类无机碳利用机理的研究

为了认识海洋浮游藻类在碳充足和碳受限条件下对水体中溶解无机碳(DIC)的利用方式与可能机理,对13种海洋浮游藻类在不同pH和CO2浓度及不同DIC条件下细胞外碳酸酐酶(CA)的活性进行了分析测定.结果显示:13种藻中,只有Amphidinium carterae和Prorocentrum minimum在碳充足条件下具细胞外CA活性.Melosira sp.、Phaeodactylum tricornutum、Skeletonema costatum、Thalassiosira rotula、Emiliania huxleyi和Pleurochrysis carterae则在碳受限条件下才具细胞外CA活性.Chaetoceros compressus、Glenodinium foliaceum、Coccolithus pelagicus、 Gephrocapsa oceanica和Heterosigma akashiwo即使在碳受限条件下也未检测到细胞外CA活性.应用封闭系统中pH漂移技术和阴离子交换抑制剂4′4′-diisothiocyanatostilbene-2,2-disulfonic acid (DIDS)等的研究表明,Coc. pelagicus和G. oceanica可通过阴离子交换机制进行HCO-3的直接利用.H. akashiwo没有潜在的HCO-3直接利用或细胞外CA催化的HCO-3利用.     

The regulation of photosynthetic rate and activation of extracellular carbonic anhydrase under CO2-limiting conditions in the marine diatom Skeletonema costatum

In the marine diatom Skeletonema costatum , carbonic anhydrase activity exterior to the plasma membrane (CA_ext) was detected only when the available CO₂ concentration was less than 5·0 mmol m^–3, this activity being unaffected by the total dissolved inorganic carbon concentration. The inhibition of CA_ext by dextran bound sulphonamide (DBS) demonstrated the key role of this enzyme in maintaining photosynthetic rate under CO₂-limited conditions. Treatment with trypsin followed by affinity chromatography on p-aminomethylbenzene-sulphamide agarose and subsequent SDS-PAGE analysis revealed a polypeptide from carbon-replete cells of identical molecular mass to the CA_ext released by trypsin from CO₂-limited cells. Redox activity in the plasma membrane of intact cells was measured by following the light-dependent reduction of ferricyanide or NADP, the greatest activity being shown by CO₂-limited cells. Overall the results suggest that high rates of redox activity under conditions of CO₂-limitation were required for the activation of CA_ext.     

Light-Induced Polar pH Changes in Leaves of Elodea canadensis: II. Effects of Ferricyanide: Evidence for Modulation by the Redox State of the Cytoplasm

The effect of an extracellular electron acceptor, ferricyanide, on the light-induced polar leaf pH changes of the submerged angiosperm Elodea canadensis in light and in darkness was determined. The rate of transmembrane ferricyanide reduction was stimulated by increased light intensity and was inhibited by inorganic carbon, indicating that changes in the redox state of the chloroplast were reflected at the plasma membrane. The addition of ferricyanide inhibited the light-induced polar leaf pH reaction. This effect could be balanced by increasing the light intensity. In the dark, the acidification induced by ferricyanide was not influenced by diethylstilbestrol at concentrations that completely inhibited the polar leaf pH changes. This indicates that the ferricyanide-induced H⁺ extrusion and the H⁺ transport during the polar reaction were mediated by different mechanisms.     

Membrane enzymes associated with the dissimilation of some citric acid cycle substrates and production of extracellular oxidation products in chemostat cultures of Pseudomonas fluorescens

Enzyme activities forming extracellular products from succinate, fumarate, and malate were examined using washed cell suspensions of Pseudomonas fluorescens from chemostat cultures. Membrane-associated enzyme activities (glucose, gluconate, and malate dehydrogenases), producing large accumulations of extracellular oxidation products in carbon-excess environments, have previously been found in P. fluorescens. Investigations carried out here have demonstrated the presence in this microorganism of a malic enzyme activity which produces extracellular pyruvate from malate in carbon-excess environments. Although the three membrane dehydrogenase enzymes decrease significantly in carbon-limited chemostat cultures, malic enzyme activity was found to increase fourfold under these conditions. The regulation of malate dehydrogenase and malic enzyme by malate or succinate was similar. Malate dehydrogenase increased and malic enzyme decreased in carbon-excess cultures. The opposite effect was observed in carbon-limited cultures. When pyruvate or glucose was used as the carbon source, malate dehydrogenase was regulated similarly by the available carbon concentration, but malic enzyme activity producing extracellular pyruvate was not detected. While large accumulations of extracellular oxalacetate and pyruvate were produced in malate-excess cultures, no extracellular oxidation products were detected in succinate-excess cultures. This may be explained by the lack of detectable activity for the conversion of added external succinate to extracellular fumarate and malate in cells from carbon-excess cultures. In cells from carbon-limited (malate or succinate) cultures, very active enzymes for the conversion of succinate to extracellular fumarate and malate were detected. Washed cell suspensions from these carbon-limited cultures rapidly oxidized added succinate to extracellular pyruvate through the sequential action of succinate dehydrogenase, fumarase, and malic enzyme. Succinate dehydrogenase and fumarase activities producing extracellular products were not detected in cells from chemostat cultures using pyruvate or glucose as the carbon source. Uptake activities for succinate, malate, and pyruvate also were found to increase in carbon-limited (malate or succinate) and decrease in carbon-excess cultures. The role of the membrane-associated enzymes forming different pathways for carbon dissimilation in both carbon-limited and carbon-excess environments is discussed.     

Impermeant electron acceptors and donors to the plasma membrane-bound respiratory chain of intact cyanobacterium Anacystis nidulans

Membrane-impermeant redox compounds ferricyanide and horse heart ferrocytochrome c acted as electron acceptor and donor, respectively, for intact cells or spheroplasts of Anacystis nidulans (Synechococcus ATCC 27144) in the dark. The anaerobic reduction of ferricyanide was faster than aerobic reduction. KCN significantly enhanced the reaction under aerobic conditions. Light did not influence ferricyanide reduction. The oxidation of exogenous ferrocytochrome c was oxygen-dependent and inhibited by KCN. Either type of redox reaction was accompanied by vectorial proton translocation out of the cells. Arrhenius plots for the temperature dependence of both ferricyanide reduction and cytochrome c oxidation gave one distinct break point reflecting the lipid phase transition temperature of the plasma membrane. The results are presented as evidence for a respiratory chain in the plasma membrane of A. nidulans.     

Plant Defense Response to Fungal Pathogens (II. G-Protein-Mediated Changes in Host Plasma Membrane Redox Reactions)

Elicitor preparations containing the avr5 gene products from races 4 and 2.3 of Cladosporium fulvum, and tomato (Lycopersicon esculentum L.) cells containing the resistance gene Cf5 were used to investigate the involvement of redox processes in the production of active oxygen species associated with the plant response to the fungal elicitors. Here we demonstrate that certain race-specific elicitors of C. fulvum induced an increase in ferricyanide reduction in enriched plasma membrane fractions of tomato cells. The addition of elicitors to plasma membranes also induced increases in NADH oxidase and NADH-dependent cytochrome c reductase activities, whereas ascorbate peroxidase activity was decreased. These results suggest that changes in the host plasma membrane redox processes, transferring electrons from reducing agents to oxygen, could be involved in the increased production of active oxygen species by the race-specific elicitors. Our results also show that the dephosphorylation of enzymes involved in redox reactions is responsible for the race-specific induced redox activity. The effects of guanidine nucleotide analogs and mastoparan on the activation of plasma membrane redox reactions support the role of GTP-binding proteins in the transduction of signals leading to the activation of the defense response mechanisms of tomato against fungal pathogens.