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【目的】从南海柳珊瑚共附生放线菌的次生代谢产物中寻找具有抗菌和抗附着活性的先导化合物。【方法】应用化学与生物活性相结合的筛选方法,从柳珊瑚共附生微生物中筛选获得代谢产物丰富且具有生物活性的目标菌株并通过大发酵提取浸膏,利用硅胶柱色谱、凝胶柱色谱和高效液相色谱等方法对发酵产物进行分离、纯化,运用波谱解析鉴定化合物的结构。【结果】从采自海南三亚的柳珊瑚(Muricella flexuosa)样品中分离到一株放线菌SCSGAA0009,鉴定为链霉属Streptomycessp.,从其改良ISP2发酵液中分离到新化合物N-(2-(1H-indol-3-yl)ethyl)propionamide(1)和已知化合物phenazine-1-carboxylic acid(2),其中化合物2对大肠杆菌和海洋细菌假单胞菌(Pseudoaltermonas piscida)具有较好抗菌活性,且有强抗草苔虫(Bugulaneritina)幼虫附着活性。【结论】首次从柳珊瑚共附生放线菌的次生代谢产物中获得新的生物碱化合物1,首次报道化合物2的抗海洋细菌活性和抗附着活性;从南海柳珊瑚共附生微生物的次生代谢产物中可以得到新化合物和活性化合物,这一来源的微生物资源值得深入研究。 相似文献
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我国资源植物化学与天然产物化学基础研究的现状与发展 总被引:8,自引:0,他引:8
本文从生物活性成分的筛选与分离、植物次生代谢产物生物合成及其分子调控、环境因子对植物次生代谢产物合成和积累的影响、植物体内生菌与植物次生代谢产物的关系等方面介绍了我国资源植物化学与天然产物化学领域基础研究的现状与发展。 相似文献
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天蓝色链霉菌全基因组序列的公布 ,对目前工业生产生物活性代谢产物菌株的遗传改造 ,构建生产高价值药物的超级菌 ,以及由微生物资源去寻找新的生物活性代谢产物将产生巨大影响。文中就基因组时代如何发展由基因组信息和化合物库预测次生代谢路径、研究功能基因组学时代的放线菌次生代谢调控、基因工程技术在放线菌抗生素生产中的应用以及体外分子定向进化与分子育种等生物技术问题进行文献综述。 相似文献
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海洋真菌及其活性代谢产物研究进展 总被引:9,自引:0,他引:9
微生物生理活性物质对人类健康极其重要。近年来,海洋微生物研究逐步成为热点,而海洋真菌是海洋微生物的重要组成部分。当前,具有生物活性的海洋真菌代谢产物被广泛应用于食品和医药产业中,研究和开发海洋真菌的生物活性代谢产物具有重要的实际意义。 相似文献
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喜树内生真菌的分离及其抗肿瘤活性代谢产物的筛选方法 总被引:16,自引:0,他引:16
从喜树(Camptotheca acuminata Decne.)的根、枝条、叶和果实中分离纯化了48株内生真菌,通过对各个菌株的少量发酵培养,HPLC分析结合色谱峰紫外扫描检测方法,以紫外扫描图谱的相似性为依据,对喜树内生真菌产生喜树碱结构类似物进行初步筛选,并进一步以抑瘤实验确证其抗肿瘤活性.结果证明,以该方法筛选到的10个内生菌株中有7个菌株发酵液对HL-60细胞增殖具有显著的抑制活性.相对于常规的生物活性筛选,高效液相色谱结合色谱峰紫外光谱的方法,在药用植物内生真菌活性次生代谢产物筛选研究中具有快速、高效的特点. 相似文献
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Kishimoto K Urade R Ogawa T Moriyama T 《Biochemical and biophysical research communications》2001,281(3):657-662
A simple method for separation and quantification of neutral lipids was developed using thin-layer chromatography (TLC) and high-performance fluorescent scanning. Neutral lipid classes were separated using the double-developing TLC method and detected by rhodamine 6G and a laser-excited fluorescent scanner. The amount of lipids applied correlated with scanned intensity volume in a dose-dependent manner. The mass of each neutral lipid band was determined by comparing band intensities of unknown samples to dilution curves of authentic standards. After scanning the dye-sprayed TLC, acyl chain species of triglyceride (TG) extracted from TLC could be determined by gas chromatography. Using this method, we quantified the amounts of TG in mouse liver and found that the measured total mass of TG correlated with that obtained by enzymatic methods. Our method should provide the basic technique for "lipidome" analysis, designed to determine and compare total lipid classes and mass present in biological samples. 相似文献
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Ondra P Zedníková K Matlach R 《Biomedical papers of the Medical Faculty of the University Palacky, Olomouc, Czechoslovakia》2005,149(2):469-472
Authors discuss problems with identification and determination of flunitrazepam and zolpidem in biological material (BM). Over the recent years, these two structurally different substances have become the most frequently used as well as abused hypnotic drugs. This study presents applicability of immunochemical methods in the screening of flunitrazepam, one of the most commonly prescribed drugs among the benzodiazepines. Herein described techniques, a liquid-liquid (L-L) extraction, solid phase extraction (SPE) and the so-called "freeze out" method are used for isolation of the above mentioned compounds from BM. Besides the thin layer chromatography (TLC) and gas chromatography - mass spectrometry (GC-MS) applied in qualitative analysis, the study also describes a gas chromatography with electron capture detector (GC-ECD) and gas chromatography with nitrogen phosphorus detector (GC-NPD) optimized for the determination of flunitrazepam and zolpidem in blood (serum). Successful analyses of these two substances are of major importance, especially in interpreting the results of forensic toxicological examinations. 相似文献
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为了建立天麻胶囊中主要有效成分天麻素的快速鉴定方法,根据天麻素的理化特性,使用乙醇和甲醇提取天麻胶囊中的天麻素,使用薄层色谱法进行鉴定,并与高效液相色谱法的分析结果进行比较。结果表明,薄层色谱法的鉴定结果与高效液相色谱法的检测结果一致,能较准确地鉴别天麻胶囊的真伪。本研究结果表明薄层色谱法能快速简便、准确灵敏地检测天麻胶囊中的有效成分,可作为法定鉴定方法的补充,对天麻胶囊实施快速初筛。 相似文献
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Guillermo E. Napolitano Jacqueline E. Richmond Arthur J. Stewart 《Soil & Sediment Contamination》1998,7(6):709-724
Petroleum hydrocarbons from 20 soils from refineries or other industrial sites were extracted with a mixture of chloroform and methanol (1:1, v/v), and the extracts were analyzed by thin layer chromatography with flame ionization detection (TLC/FID). The TLC/FID procedure has been used widely in biological and medical research but generally has been underutilized in environmental chemistry. The analysis method involved spotting a small volume of sample extract (typically 1 to 3?µl) on ten silica-coated quartz rods, and chromatographically separating constituents in the spots using solvent systems of increasing polarities (hexane, toluene, and dichloromethane + methanol). We achieved complete separation of saturated hydrocarbons, aromatic hydrocarbons, resins, and asphaltenes from the hydrocarbon-contaminated soils with this method. Analysis of the separated constituents by TLC/FID also allowed quantification of aromatic and aliphatic hydrocarbons without interference from soil biogenic lipids. A simplified version of the method permitted excellent separation of aliphatics +aromatics (forming a single peak) from resins and asphaltenes. The procedure is rapid (complete analysis of ten samples in about 1?h after extraction). Thus, the method seems well suited for synoptic surveys or screening and characterizing numerous samples prior to using more detailed and costly analyses. 相似文献
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Maciej Jaskiewicz Malgorzata Orlowska Gabriela Olizarowicz Dorian Migon Daria Grzywacz Wojciech Kamysz 《International journal of peptide research and therapeutics》2016,22(2):155-161
Increasing resistance to conventional antibiotics among microorganisms is one of the leading problems of medicine nowadays. Antimicrobial peptides are compounds exhibiting both antibacterial and antifungal activities. However, it is difficult to predict whether a designed new compound would exhibit any biological activity. Moreover, purification of the peptides is one of the most time-consuming and expensive steps of the synthesis that sometimes leads to unnecessary loss of solvents and reagents. In our study we have developed a thin-layer chromatography (TLC) direct bioautography technique for rapid determination of antimicrobial activity of peptides without the necessity of high-performance liquid chromatography purification. In this assay, crude peptides were applied and separated on a TLC plate. Then, pre-prepared plates were dipped into microbial suspension and incubated under optimum conditions for bacteria and fungi as well. The activity of the tested compounds was visualized by spraying the TLC plates with a cell viability reagent, resazurin (7-hydroxy-3H-phenoxazin-3-one 10-oxide). Effectiveness of this assay was compared with minimal inhibitory concentration results obtained by broth microdilution assay. Interestingly, so far such a screening method has not been applied for this group of compounds. 相似文献
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JaspricaI Smolcic-BubaloA MomarA Medic-SaricM 《天然产物研究与开发》2004,16(4):302
Flavonoids and phenolic acids with a variety of biological activity are considered to be the main compounds in propolis - anatural product produced by the honey bee. TLC can be used for rapid screening of pharmacologically active components and to establish the difference between different propolis samples. Char goal was to optimize chromatographic conditions for separation of flavonoids and phenolic acids and to apply the optimized method for analysis of propolis samples from different geographic regions of Croatia. For chromatographic analysis we used 20 cm x 20 cut glass-backed TLC plates coated with 0.25 mm layers of silica gel 60 F-254.Ethanolic standard solutions (80%) of the flavonoids and phenolic acids (10 muL) were applied to the plates. Chromatograms were developed at room temperature by ascending development in previously saturated vertical, flat-bottomed glass chambers with glass lids. Visualization was performed in short- and long-wavelength UV light and in long-wavelength UV light after spraying with different reagents. After calculation of R F values numerical taxonomy methods were used to test the efficiency of 11 mobile phases and to optimize chromatographic conditions for separation of 19 standard solutions. We established the most appropriate mobile phases (chloroform-methanol-(98-100% ) formic acid, 44.1 3 2.35, and n-hexane-ethyl acetate-glacial acetic acid, 31 14 5) for separation of standards. The results obtained were used for analysis of propolis samples. TLC was shown to be a highly suitable method for rapid analysis of propolis samples. It can be used to establish differences between the amounts of pharmacologically active compcunds in propolis from different geographic regions of Croatia. 相似文献
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Imran Sajid Clarisse Blandine Fotso Fondja Yao Khaled Attia Shaaban Shahida Hasnain Hartmut Laatsch 《World journal of microbiology & biotechnology》2009,25(4):601-610
A culture collection of 110 indigenous Streptomyces strains originally isolated from saline farmlands (Punjab, Pakistan) using stringent methods was screened biologically and
chemically to investigate their potential for the production of bioactive secondary metabolites. In a biological screening
the crude extracts obtained from the culture broth of selected strains were analysed for their activity against a set of test
organisms, including Gram-positive, Gram-negative bacteria, fungi and microalgae using the disk diffusion bioassay method.
Additionally a cytotoxicity test was performed by means of the brine shrimp microwell cytotoxicity assay. In a chemical screening
each of the crude extracts was analysed by TLC using various staining reagents and by HPLC-MS/MS measurements. The results
depicted an impressive chemical diversity of crude extracts produced by these strains. The taxonomic status of the selected
strains was confirmed by preliminary physiological testing and 16S rRNA gene sequencing. 相似文献
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In the course of screening for nitric oxide inhibitors in activated microglial BV-2 cells, cyclo(dehydrohistidyl-Ltryptophyl) was isolated from solid-state fermentation cultures of an unidentified fungal strain, Fb956. Its structure was determined by spectroscopic methods including 2D NMR and chiral TLC analyses. Cyclo(dehydrohistidyl-L-tryptophyl) was found to have an inhibitory activity on nitric oxide production with an IC50 of 6.5 muM in activated BV-2 cells. The structure determination and biological activity of cyclo(dehydrohistidyl- L-tryptophyl) was reported for the first time in this study. 相似文献
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P Arun A Ravikumar S Leelamma P A Kurup 《Indian journal of biochemistry & biophysics》1998,35(5):308-312
A procedure for estimation of digoxin in biological samples after adding a known quantity of digoxin followed by extraction, separation by TLC and HPLC is described. The identity of digoxin thus extracted from rat brain has been established by reaction with digoxin antibody and by its inhibition of Na(+)-K+ ATPase activity. The method could be a better substitute to the routine radioimmunoassay as interfering substances are removed by TLC and HPLC. 相似文献
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A simple TLC-densitometric technique has been developed for the rapid and accurate analysis of artemisinin in a large number of Artemisia annua plantlets cultured in vitro. This new analytical method is based on the structural conversion of artemisinin on a silica gel layer by ammonia vapour to form 10-azadesoxyartemisinin, a chromophore-containing compound (lambdamax 320 nm) that can be detected by UV-based TLC densitometry. The TLC system was evaluated quantitatively in terms of product stability, precision, accuracy and calibration. Good linearity was obtained in the range of 0.01-0.12 microg artemisinin. The technique appeared to be accurate and sensitive as compared with the complicated pre-column reaction-HPLC technique. Among 90 samples of A. annua plantlets, the artemisinin content in the leaves appeared to be highly variable, ranging from 0.02 to 0.67% w/w dry weight. These results demonstrate that densitometric TLC can be a cheap and simple technique for the accurate screening of high-artemisinin-producing plants. 相似文献