长裙竹荪抑菌活性物质的分离纯化及其抑菌效果

长裙竹荪Dictyophora indusiata是珍贵的食药用真菌,具有很强的抑菌作用,在天然防腐剂开发方面具有广阔的应用前景。本研究以长裙竹荪的抑菌活性为指标,通过萃取、3次不同流动相的硅胶柱层析、1次反相柱层析和薄层层析法对竹荪提取物进行分离纯化,得到一个抗菌活性强的单体化合物。根据核磁共振波谱等数据分析,推断该化合物为间苯三酚。以巨大芽孢杆菌和肠炎沙门氏菌为供试菌,用平板打孔法及原位抑菌法测定该化合物的抑菌效果,结果表明：该化合物对这两种菌有很强的抑制作用,半抑制浓度分别为83.06μg/mL和51.58μg/mL。本研究首次从长裙竹荪中获得具有抗菌活性的单体化合物间苯三酚,为竹荪天然抗菌物质的开发提供理论依据。     

Abnormal deoxyribose metabolites in the urine of a child with a possible new inborn error of metabolism

The urinary extract of a child investigated because of strabismus was found to contain large amounts of a compound which was identified using gas chromatography/mass spectrometry as 2-deoxyerythropentono-1,4-lactone. This lactone has not been observed previously in urinary extracts. When ion-exchange chromatography was used to isolate the organic acids from urine, the major peaks obtained by gas chromatography were shown to be 2-deoxyerythropentonic acid, 2-deoxyerythropentono-1,5-lactone and 2-deoxyerythropentono-1,4,lactone. Another abnormal metabolite, 2-deoxyribitol, was also excreted by the patient although this compound could not be detected in the urine of normal children. It is proposed that these unusual compounds accumulate in the urine of this child as a result of a defect in the catabolism of 2-deoxyribose.     

Identification of the Basic Structure of a Glycolipid from Selenomonas ruminantium as β-Glucosaminyl-1,6-glucosamine

Lipid materials extracted by chloroform-methanol from solvent and acid treated cells of Selenomonas ruminantium grown with ¹⁴C-valerate were analyzed by thin-layer chromatography. At least 12 radioactive lipid materials were present in the extract. The major compound, designated as spot A, carried approximately 70% of radioactivity of this fraction. The spot A compound was purified by column and thin-layer chromatography and its chemical structure was studied. The basic structural unit of this material was tentatively identified as β-gIucosaminyl-1,6-glucosamine with O-acyl and amide linked fatty acids.     

High concentration of free trimethyllysine in red blood cells

A high concentration of a basic unidentified amino compound was found in the blood of rats. It was isolated and identified as N epsilon,N epsilon,N epsilon-trimethyllysine by paper chromatography, thin-layer chromatography, high-performance liquid chromatography and amino acid analyzer. It was localized exclusively in red blood cells in the blood of rats. Free trimethyllysine was also determined in the liver, kidney, spleen, brain, muscle, heart and testis of rat. The concentration of free trimethyllysine in red blood cells was more than 10-times as high as that in the other tissues. This compound in red blood cells was found in different species of animals. The relationship between this free trimethyllysine and carnitine was discussed.     

微生物来源的HLE抑制剂N01WA-735E的研究

人白细胞弹性蛋白酶抑制剂为筛选炎症和癌症的重要靶点。应用白细胞弹性蛋白酶抑制剂高通量的筛选模型对数千株放线菌进行筛选,发现了阳性菌株N01 WA-735。首先通过形态学和化学分类学鉴定其为链霉菌属。采用有机溶剂提取、硅胶柱色谱、Sephadex LH-20柱色谱和结晶等方法对该菌株的发酵产物进行了分离纯化,得到活性单体化合物N01 WA-735E,通过对N01 WA-735E的理化性质和波谱数据分析,确定其结构与文献报道的化合物BE-52440A相同。该化合物对人白细胞弹性蛋白酶有很强的抑制活性,其IC50为3.02μmol/L。该化合物对人白细胞弹性蛋白酶的抑制活性国内外未见报道。     

Isolation and identification of a small molecular weight inhibitor of cyclic nucleotide phosphodiesterase from bovine brain.

In a search for endogenous regulators for cyclic nucleotide phosphodiesterase (3':5'-cyclic-AMP 5'-nucleotidohydrolase, EC 3.1.4.17), we found that the ultrafiltrate of bovine brain homogenate contained a cyclic nucleotide phosphodiesterase inhibitor. The inhibitor-containing fraction was further purified by ion-exchange column chromatography and gel filtration chromatography. The purified inhibitor was found to be a small molecular weight compound which had a maximum absorption at 248 nm. This compound was identified by thin-layer chromatography and high-pressure liquid chromatography as hypoxanthine. We suggest that hypoxanthine may serve as an endogenous regulator for the hydrolysis of cyclic nucleotide by cyclic nucleotide phosphodiesterase.     

Identification and quantification of 8,5'-cyclo-2'-deoxy-adenosine in DNA by liquid chromatography/ mass spectrometry

Recent studies suggested that 8,5'-cyclo-2'-deoxyadenosine may play a role in diseases with defective nucleotide-excision repair. This compound is one of the major lesions, which is formed in DNA by hydroxyl radical attack on the sugar moiety of 2'-deoxyadenosine. It is likely to be repaired by nucleotide-excision repair rather than by base-excision repair because of a covalent bond between the sugar and base moieties. We studied the measurement of 8,5'-cyclo-2'-deoxyadenosine in DNA by liquid chromatography/isotope-dilution mass spectrometry. A methodology was developed for the analysis of 8,5'-cyclo-2'-deoxyadenosine by liquid chromatography in DNA hydrolyzed to nucleosides by a combination of four enzymes, i.e., DNase I, phosphodiesterases I and II, and alkaline phosphatase. Detection by mass spectrometry was performed using atmospheric pressure ionization-electrospray process in the positive ionization mode. Results showed that liquid chromatography/isotope-dilution mass spectrometry is well suited for identification and quantification of 8,5'-cyclo-2'-deoxyadenosine in DNA. Both (5'R)- and (5'S)-diastereomers of 8,5'-cyclo-2'-deoxyadenosine were detected. The level of sensitivity of liquid chromatography/mass spectrometry with selected-ion monitoring amounted to 2 fmol of this compound on the column. The yield of 8,5'-cyclo-2'-deoxyadenosine was measured in DNA in aqueous solution exposed to ionizing radiation at doses from 2.5 to 80 Gray. Gas chromatography/mass spectrometry was also used to measure this compound in DNA. Both techniques yielded similar results. The yield of 8,5'-cyclo-2'-deoxyadenosine was comparable to the yields of some of the other major modified bases in DNA, which were measured using gas chromatography/mass spectrometry. The measurement of 8,5'-cyclo-2'-deoxyadenosine by liquid chromatography/mass spectrometry may contribute to the understanding of its biological properties and its role in diseases with defective nucleotide-excision repair.     

Dibutyl phthalate, the bioactive compound produced by Streptomyces albidoflavus 321.2

It was found that the bioactive compound, dibutyl phthalate, was produced by a new soil isolate Streptomyces albidoflavus 321.2. Once this active compound was recovered by ethyl acetate from the fermented broth, being possible to isolate 13.4 mg/l, it was purified by paper, silica gel column, thin layer and gas chromatography. Structure was determined by analysing UV, IR and GC-MS spectra. During analysis, such active compound showed strong activity against gram-positive and gram-negative bacteria, as well as unicellular and filamentous fungi. The antimicrobial activity of the compound was reversed by the amino acid proline. No acute toxicity was observed.     

中国被毛孢发酵液中一种镇静催眠活性物质的分离纯化和结构鉴定

本研究对中国被毛孢Hirsutella sinensis菌株RCEF0273发酵液的乙酸乙酯粗提物中的镇静催眠活性物质进行了分析.通过活性指导下的色谱分离,从乙酸乙酯提取物中得到了一种白色晶体化合物,质谱(MS)和核磁共振(NMR)的鉴定结果表明该化合物为茶碱-9-葡萄糖苷.活性试验显示该化合物可抑制小鼠的自发活动,缩短小鼠入睡潜伏期,延长小鼠睡眠持续时间,具有镇静催眠活性.     

Dihydroconiferyl alcohol — A cell division factor from Acer species

A compound that stimulated growth of soybean callus was isolated from spring sap of sycamore (Acer pseudoplatanus L.). Insufficient compound was isolated to permit it to be characterised. A compound with identical properties was isolated from commercial maple syrup, the concentrated spring sap of Acer saccharum L. The compound was identified as 3-(3-methoxy-4-hydroxyphenyl)-propan-1-ol (dihydroconiferyl alcohol, DCA). DCA was also active in the tobacco callus and radish leaf senescence assays, but was inactive in four other tests for cytokinin activity. DCA acted synergistically with kinetin to promote soybean callus growth. It is concluded that DCA has properties distinct from those of purine cytokinins.Abbreviation DCA dihydroconiferyl acohol - GC gas chromatography - IAA indole-3-acetic acid - iP isopentenyladenine - [9R]iP isopentenyladenosine - LC liquid chromatography - MS mass spectrometry - NAA 1-napthylacetic acid - TLC thinlayer chromatography - TMSi trimethylsilyl     

Fractionation and isolation of the multiple forms of prorennin (prochymosin)

The heterogeneity of prorennin was studied by chromatography on DEAE-cellulose and microgranular DEAE-cellulose columns, as well as by polyacrylamide-gel electrophoresis. Prorennin prepared by alum treatment, salting-out and chromatography was resolved into three components by a compound gradient of sodium phosphate on microgranular DEAE-cellulose. Polyacrylamide-gel electrophoresis confirmed the chromatographic results, but crystalline rennin was shown to consist of four bands. When prorennin was isolated directly by chromatography, four zymogen components were resolved on microgranular DEAE-cellulose with a modified compound gradient of sodium phosphate. Polyacrylamide-gel electrophoresis confirmed the existence of four multiple forms of prorennin as well as homogeneity of the chromatographic fractions.     

Identification of aminoethylcysteine ketimine decarboxylated dimer in human plasma

Aminoethylcysteine ketimine decarboxylated dimer (AECK-DD) is a natural sulfur-containing tricyclic compound detected, until now, in human urine and bovine cerebellum. Recently, the antioxidant properties of this compound, and particularly its protective effect on the in vitro oxidation of low-density lipoproteins, have been demonstrated. In this paper, the identification of AECK-DD in human plasma by means of gas chromatography, high-performance liquid chromatography and gas chromatography–mass spectrometry, performed after a simple and fast purification procedure, is reported.     

Structure of Xanthocidin

A burnt flavoring compound, which imparts to aged sake its characteristic and dominant flavor, was isolated by Diaion HP–20, silicic acid and Dowex 1–X8 (CH₃COO^?) column chromatography and chloroform extraction. Based on thin-layer and gas liquid chromatography, UV and GC–MS spectral data, it was identified as 3-hydroxy-4,5-dimethyl-2 (5H)-furanone and its structure was also confirmed by synthesis. It was suggested that this compound was formed by the condensation of a-ketobutyric acid with acetaldehyde which occurred from degradation of threonine.     

Detection of 2H-1,4-thiazine-5,6-dihydro-3-carboxylic acid (aminoethylcysteine ketimine) in the bovine brain

2H-1,4-Thiazine-5,6-dihydro-3-carboxylic acid (trivial name: aminoethylcysteine ketimine) is a cyclic sulfur-containing imino acid detected in bovine brain extracts by means of three different procedures. Gas liquid chromatography of protein-free extracts of five bovine brains revealed the presence of this compound at concentrations ranging from 2 to 3 nmol/g wet weight of tissue. The enzymatic method based on the inhibition of D-amino acid oxidase activity by aminoethylcysteine ketimine together with an high-performance liquid chromatography procedure confirm the identification and quantitations obtained with gas liquid chromatography. The discovery of this compound structurally similar to pipecolic acid opens the question of its physiological role in the central nervous system.     

Determination of peptido-aminobenzophenone (2-o-chlorobenzoyl-4-chloro-N-methyl-N′-glycyl-glycinanilide) by electron-capture gas—liquid chromatography

A gas—liquid chromatographic method for the determination of peptido-aminobenzophenone (2-o-chlorobenzoyl-4-chloro-N-mehtyl-N′-glycyl-glycinanilide, I) in dog plasma was developed. Decomposition of compound I was observed during chromatography. In alkaline medium, compound I in plasma was submitted to ring closure to yield 3-amino-6-chloro-5-(2-chlorophenyl)-1-methylquinolin-2-one (aminoquinolone), and the hexane extract was assayed by gas—liquid chromatography using electron-capture detection. The concentration range of compound I studied was 10–90 ng per 0.5 ml of plasma. Interference from both endogenous and exogenous sources was negligible. The method could be applied to the determination of the plasma level of compound I in dogs following oral administration of a single 5 mg/kg dose.     

一种白僵菌中MAO抑制剂的分离纯化和结构鉴定

本研究对前期筛选出的一株具有较强的单胺氧化酶(MAO)抑制活性的白僵菌菌株Ba02进行了液体培养;通过不同提取剂的提取效果比较,发现乙酸乙酯能较好地提取出该发酵液中单胺氧化酶抑制剂。通过活性指导下的色谱分离,从乙酸乙酯提取物中得到了一种深红色粉末状化合物。活性测定结果显示该化合物在15μg.mL-1时对MAO-A和MAO-B的抑制率分别为97.50%和95.34%。MS和NMR的鉴定结果表明该化合物为卵孢菌素(Oosporein)。虽然该化合物是一已知化合物,但其对单胺氧化酶的抑制活性尚属首次发现。     

Chemical behaviour of cytidine 5'-monophospho-N-acetyl-beta-D-neuraminic acid under neutral and alkaline conditions

The chemical behaviour of CMP-N-acetylneuraminic acid under neutral and different alkaline conditions has been investigated. The products formed were isolated by ion-exchange chromatography and gel filtration and analysed by colorimetric methods, thin-layer chromatography, combined gas-liquid chromatography/mass spectrometry and/or 360-MHz 1H-NMR spectroscopy. A maximum stability of CMP-N-acetylneuraminic acid was observed at pH8-11. In the tested pH range of 6-13, CMP and N-acetylneuraminic acid were formed in variable amounts as decomposition products. 2-Deoxy-2,3-dehydro-N-acetylneuraminic acid was produced at pH greater than 7; the amount of this substance increased with increasing pH. In anhydrous triethylamine its yield was 50%. A new neuraminic acid derivative, N-acetyl-beta-D-neuraminic acid 2-phosphate, could be isolated from the mixture of alkaline decomposition products of CMP-N-acetylneuraminic acid. The yield of this compound was maximum 22% in anhydrous triethylamine. Because 2-deoxy-2,3-dehydro-N-acetylneuraminic acid was formed under simulated physiological conditions, it is assumed that this compound, which occurs in tissues and fluids of man and animals, is derived from CMP-N-acetylneuraminic acid non-enzymically also under conditions in vivo.     

N tau-Ribosylhistidine, a novel histidine derivative in urine of histidinemic patients. Isolation, structure, and tissue level

On amino acid analysis of urine of histidinemic patients, an unidentified compound was eluted in a position between beta-aminoisobutyric acid and gamma-aminobutyric acid. This compound was purified to homogeneity from the urine by a combination of extraction with 80% ethanol, repeated column chromatography on Bio-Rad AG-50, and high performance liquid chromatography on a strongly cationic ion exchanger. The compound yielded free histidine on hydrolysis in an evacuated sealed tube with 0.1-6.0 M HCl at 145 degrees C for 5 h, but not at 100 degrees C for 24 h. This compound was determined to be N tau-ribosylhistidine by 1H and 13C NMR spectroscopies. The urinary content of this material in normal and histidinemic children was 17.8 +/- 13.4 (n = 10) and 126 +/- 51 (n = 14) mumol/g creatinine (mean +/- S.D.), respectively, and were closely correlated with those of urinary histidine. The renal clearance value of N tau-ribosylhistidine in humans was 96% of that of creatinine. When rats were fed on diets rich in histidine, the urinary excretion of N tau-ribosylhistidine increased greatly and was well correlated with the intake of histidine.     

Detection and characterization of the Gloeosporium gloeosporioides growth inhibitory compound iturin A from Bacillus subtilis strain KS03

The Bacillus subtilis strain KS03 was isolated, and identified as a biological control agent that inhibits the anthracnose disease fungus Gloeosporium gloeosporioides. The antifungal compound was purified from its culture broth through butanol extraction, diethylaminoethyl (DEAE) Sepharose CL-6B chromatography, and preparative thin layer chromatography. Tandem mass spectrometric analyses (MS/MS), with matrix-assisted laser desorption ionization (MALDI) time-of-fight/time-of-flight (TOF/TOF) mass spectrometry, showed that the antifungal compound was iturin A, a cyclic lipopeptide antibiotic. The major compound, with a molecular mass of 1042 Da, was identified as iturin A(2).     

新疆蓝刺头化学成分研究

以新疆蓝刺头(Echinops ritro L.)全草为研究材料,通过硅胶柱色谱,Sephadex LH-20柱色谱,重结晶等技术对新疆蓝刺头化学成分进行分离纯化,通过理化性质分析及1H-NMR,13C-NMR等技术对化合物结构进行鉴定.结果表明,共分离出5个化合物,分别是三萜类化合物蒲公英甾醇乙酰酯(化合物1)、蒲公英甾醇(化合物2)、黄酮苷类化合物金丝桃苷(化合物3)、胡萝卜苷(化合物4)与β-豆甾醇葡萄糖苷(化合物5).其中化合物1,2,5首次从该种植物中分离,化合物3为首次从该属植物中分离.