紫叶李果实的化学成分研究

采用硅胶、ODS和Sephadex LH-20柱色谱等方法对紫叶李果实95%乙醇提取物进行提取分离纯化,运用IR、UV、1H NMR、13C NMR、HMBC、HSQC等波谱学技术鉴定5个化合物:3-O-乙酰基原儿茶酸(1)、2(R)-羟基丁二酸-1-甲酯(2)、3,3',4,4'-四羟基联苯(3)、β-胡萝卜苷(4)和槲皮素(5)。化合物1~4是首次从该植物中分离得到,其中化合物1为新天然产物,并且首次报道化合物1的1H NMR,13C NMR数据。对已分离的5个化合物进行了DPPH自由基清除实验,结果显示化合物3、5具有潜在的抗氧化活性。     

香港地钱抗菌化学成分研究

利用生物活性试验和^1H NMR追踪，从香港地钱（Marchantia paleacea Bertol)的乙醇提取物中简单快速分离具有抗菌活性的三个已知芳香化合物：2－羟基－3，7－二甲氧基菲（1），marchantin C(2)和isoriccardin C(3)。化合物（1）的NMR测定以及晶体结构对文献值部分NMR数据进行了修正。     

黄鞘蕊花中的艾里莫芬烷型倍半萜(英文)

从黄鞘蕊花 (ColeusxanthanthusC .Y .WuetY .C .Huang)的乙酸乙酯部分首次分离得到 3个艾里莫芬烷型倍半萜类化合物 ( 1～ 3) ,其结构由NMR波谱技术及单晶X衍射分析确定。其中 ,化合物 1为新化合物 ,命名为 8,9_断裂_艾里莫芬_1 ( 1 0 ) ,7( 1 1 )_二烯_8,1 2_内酯_9_酸。首次对化合物 2 ( 2 ,9_二氧代优瑞坡森 )和 3( 9_氧代宽眼菊素 )的1H NMR和13C NMR谱进行了全归属。     

木霉菌KK19L1发酵液中的一个新化合物

对菌株Trichoderma sp.KK19L1液体发酵,在该菌发酵液的乙酸乙酯层分离得到三个化合物,运用1H NMR、13C NMR和HR-ESI-MS等光谱学方法并参考相应的文献,确定其结构分别为(E)-3-acetylbenzyl but-2-enoate(1)、1-羟基-6-甲基-9,10-蒽醌(2)、2-甲基-3-羟甲基-5-羟基-7-甲氧基色原酮(3),其中化合物1为新的化合物,化合物3为首次在该菌株中分离得到。     

海洋来源真菌Hansfordia sinuosae中的一个新异香豆素类化合物

对海洋来源真菌Hansfordia sinuosae的化学成分进行研究,采用硅胶柱、Sephadex LH-20柱和半制备HPLC色谱分离纯化,运用1D NMR(1H NMR、13C NMR)、2D NMR(HMQC、1H-1H COSY、HMBC)及MS等现代波谱学方法并结合相关的文献,从中分离鉴定3个异香豆素类化合物,分别为:(E)-6,8-dihydroxy-3-(prop-1-en-1-yl)-1H-isochromen-1-one(1),(R)-6-hydroxymellein(2),3-methyl-6,8-dihydroxyisocoumarin(3)。其中,化合物1为新化合物,化合物3为首次在该种菌株中分离得到。     

鞭檐犁头尖中的苯丙素甙类化合物

从天南星科植物鞭檐犁头尖的根茎中分离得到3个化合物,运用波谱技术(UV,IR,^1HNMR,^13C NMR and 2D NMR)对它们的化学结构进行解析,分别鉴定为松柏甙(1),甲基松柏甙(2)和硝酸钾(3)。从鞭檐犁头尖中分离得到苯丙素甙类化合物尚属首次。     

羊角拗根的化学成分研究

应用柱色谱技术从羊角拗[Strophanthus divaricatus(Lour.)Hook.et Arn.]根的甲醇提取物中分离纯化出6个化合物,通过1H NMR和13C NMR等波谱技术并对照文献,鉴定其分别为(+)-pinoresinol(1)、沙门苷元(2)、17βH-沙门苷元(3)、羊角拗苷(4)、β-谷甾醇(5)和胡萝卜苷(6)。其中,化合物2~4均为强心苷类化合物。化合物1和3为首次从该植物中分得。     

北极真菌Eutypella sp.D-1中海松烷二萜类化合物的研究

采用硅胶柱、反相硅胶柱和凝胶柱等色谱技术,从北极真菌Eutypella sp.的菌体发酵液提取物中分离纯化得到2个化合物,通过1H和13C NMR、2D NMR等分析方法,并比较相关文献,鉴定这两个化合物为libertellenone C(1)和libertellenone A(2)。测试了化合物1和2对各种癌细胞株的细胞毒活性,其中化合物1对胰腺癌SW1990和胶质瘤U251有一定的细胞毒活性,此外化合物2对稻瘟霉有抑制活性。     

松针的化学成分研究

从采自四川夹江的马尾松叶(Pinus massonianaLams.)中分离得到二个化合物,经IR1、H NMR1、3C NMR、2D-NMR、MS等现代波谱技术鉴定为ent-8,13-epoxylabd-14-en-19-oic acid(1)和槲皮素(2)。均为首次从马尾松叶中分离得到。     

海洋真菌灰黄青霉Penicillium griseofulvum次级代谢产物中一个新的内酯醛结构

采用硅胶柱色谱,半制备高压液相色谱(HPLC),ODS柱色谱以及Sephadex LH-20凝胶柱色谱等多种方法从海洋真菌灰黄青霉Penicillium griseofulvum发酵液的乙酸乙酯萃取物中分离纯化得到3个化合物,采用1D NMR(1H NMR、13C NMR、DEPT)、2D NMR(HMBC、HMQC、1H-1H COSY)及MS等光谱学方法分析及参考相应的文献,确定它们的结构分别为:2-(6-羟基-5,7-二甲基苯并呋喃酮-4-基)乙醛(1)、对苯二甲酸二(2-乙基)己酯(2)和己二酸二乙基己酯(3),其中化合物1为新化合物。     

Two new cytochalasins from an endophytic fungus,KL-1.1 isolated from Psidium guajava leaves

Chemical investigation of the endophytic fungus, KL-1.1, isolated from the leaves of Psidium guajava (Linn) led to the isolation of two new cytochalasin derivatives, 18-desoxy-19,20-epoxycytochalasin C and 18-desoxycytochalasin C, together with five other known derivatives. The structures of the isolated compounds were elucidated by one and two dimensional nuclear magnetic resonance spectroscopy as well as by mass spectrometry. These compounds represent novel chemical scaffold with potential for development into anticancer agents.     

Diversity of type I polyketide synthase genes in the wood-decay fungus Xylaria sp. BCC 1067

Fungal type I polyketide (PK) compounds are highly valuable for medical treatment and extremely diverse in structure, partly because of the enzymatic activities of reducing domains in polyketide synthases (PKSs). We have cloned several PKS genes from the fungus Xylaria sp. BCC 1067, which produces two polyketides: depudecin (reduced PK) and 19,20-epoxycytochalasin Q (PK-nonribosomal peptide (NRP) hybrid). Two new degenerate primer sets, KA-series and XKS, were designed to amplify reducing PKS and PKS-NRP synthetase hybrid genes, respectively. Five putative PKS genes were amplified in Xylaria using KA-series primers and two more with the XKS primers. All seven are predicted to encode proteins homologous to highly reduced (HR)-type PKSs. Previously designed primers in LC-, KS-, and MT-series identified four additional PKS gene fragments. Selected PKS fragments were used as probes to identify PKS genes from the genomic library of this fungus. Full-length sequences for five PKS genes were obtained: pks12, pks3, pksKA1, pksMT, and pksX1. They are structurally diverse with 1-9 putative introns and products ranging from 2162 to 3654 amino acids in length. The finding of 11 distinct PKS genes solely by means of PCR cloning supports that PKS genes are highly diverse in fungi. It also indicates that our KA-series primers can serve as powerful tools to reveal the genetic potential of fungi in production of multiple types of HR PKs, which the conventional compound screening could underestimate.     

Indole alkaloids from the leaves of Philippine Alstonia scholaris

The first seco-uleine alkaloids, manilamine (1) (18-hydroxy-19,20-dehydro-7,21-seco-uleine) and N4-methyl angustilobine B (2), were isolated from the (pH 5) alkaloid extract of Philippine Alstonia scholaris leaves together with the known indole alkaloids 19,20-(E)-vallesamine (3), angustilobine B N4-oxide (4), 20(S)-tubotaiwine (5), and 6,7-seco-angustilobine B (6). The structure of the alkaloids was established from MS and NMR experiments.     

Glutamic acid residues of bacteriorhodopsin at the extracellular surface as determinants for conformation and dynamics as revealed by site-directed solid-state 13C NMR

We recorded (13)C NMR spectra of [3-(13)C]Ala- and [1-(13)C]Val-labeled bacteriorhodopsin (bR) and a variety of its mutants, E9Q, E74Q, E194Q/E204Q (2Glu), E9Q/E194Q/E204Q (3Glu), and E9Q/E74Q/E194Q/E204Q (4Glu), to clarify contributions of the extracellular (EC) Glu residues to the conformation and dynamics of bR. Replacement of Glu-9 or Glu-74 and Glu-194/204 at the EC surface by glutamine(s) induced significant conformational changes in the cytoplasmic (CP) surface structure. These changes occurred in the C-terminal alpha-helix and loops, and also those of the EC surface, as viewed from (13)C NMR spectra of [3-(13)C]Ala- and [1-(13)C]Val-labeled proteins. Additional conformational changes in the transmembrane alpha-helices were induced as modified retinal-protein interactions for multiple mutants involving the E194Q/E204Q pair. Significant dynamic changes were induced for the triple or quadruple mutants, as shown by broadened (13)C NMR peaks of [1-(13)C]Val-labeled proteins. These changes were due to acquired global fluctuation motions of the order of 10(-4)-10(-5) s as a result of disorganized trimeric form. In such mutants (13)C NMR signals from Val residues of [1-(13)C]Val-labeled triple and quadruple mutants near the CP and EC surfaces (including 8.7-A depth from the surface) were substantially suppressed, as shown by comparative (13)C NMR studies with and without 40 micro M Mn(2+) ion. We conclude that these Glu residues at the EC surface play an important role in maintaining the native secondary structure of bR in the purple membrane.     

On the biosynthesis of 19,20-dehydroprostaglandin E2

19,20-Dehydro-PGE1 and 19,20-dehydro-PGE2 were recently identified in human seminal fluid. These prostaglandins might be formed by dehydration of 19(R)-hydroxy-PGE1 and 19(R)-hydroxy-PGE2 or, conceivably, by biosynthesis from precursor fatty acids with a terminal double bond. To examine the latter possibility, 5(Z), 8(Z), 11(Z), 14(Z), 19-eicosapentaenoic acid was prepared by chemical synthesis and incubated with microsomes of ram vesicular glands and glutathione (1 mM). The fatty acid was converted to 19,20-dehydro-PGE2, which was identified by GC-MS, by UV analysis after alkali treatment and by oxidative ozonolysis. The semisynthetic 19,20-dehydro-PGE2 and the corresponding compound in human seminal fluid showed the same polarity on reversed phase HPLC and virtually identical mass spectra. The described method might be used to generate 19,20-dehydro-PGE2 for evaluation of its biological effects.     

A method for analysis of pulmonary arterial and venous occlusion data.

Recently, we presented a compartmental model of the pulmonary vascular resistance (R) and compliance (C) distribution with the configuration C1R1C2R2C3 (J. Appl. Physiol. 70: 2126-2136, 1991). This model was used to interpret the pressure vs. time data obtained after the sudden occlusion of the arterial inflow (AO), venous outflow (VO), or both inflow and outflow (DO) from an isolated dog lung lobe. In the present study, we present a new approach to the data analysis in terms of this model that is relatively simple to carry out and more robust. The data used to estimate the R's and C's are the steady-state arterial [Pa(0)] and venous [Pv(0)] pressures, the flow rate (Q), the area (A2) encompassed by Pa(t) after AO and the equilibrium pressure (Pd) after DO, and the average slope (m) of the Pa(t) and Pv(t) curves after VO. The following formulas can then be used to calculate the 2 R's and 3 C's: [Pa(0) - Pv(0)]/Q = R1 + R2 = RT, R1C1 congruent to to A2/[Pa(0) - Pd], R1 congruent to [Pa(0) - Pd]/Q, Q/m = C1 + C2 + C3 = CT, and C2 = CT - (RTC1/R2).     

Fourier transform infrared spectrum of the secondary quinone electron acceptor Q(B) in photosystem II

Fourier transform infrared difference spectra upon single reduction of the secondary quinone electron acceptor Q(B) in photosystem II (PSII), without a contribution from the electron donor-side signals, were obtained for the first time using Mn-depleted PSII core complexes of the thermophilic cyanobacterium Thermosynechococcus elongatus. The Q(B)(-)/Q(B) difference spectrum exhibited a strong C...O stretching band of the semiquinone anion at 1480 cm(-)(1), the frequency higher by 2 cm(-)(1) than that of the corresponding band of Q(A)(-), in agreement with the previous S(2)Q(B)(-)/S(1)Q(B) spectrum of the PSII membranes of spinach [Zhang, H., Fischer, G., and Wydrzynski, T. (1998) Biochemistry 37, 5511-5517]. Also, several peaks originating from the Fermi resonance of coupled His modes with its strongly H-bonded NH vibration were observed in the 2900-2600 cm(-)(1) region, where the peak frequencies were higher by 7-24 cm(-)(1) compared with those of the Q(A)(-)/Q(A) spectrum. These frequency differences suggest that H-bond interactions of the CO groups, especially with a His side chain, are different between Q(B)(-) and Q(A)(-). Furthermore, a prominent positive peak was observed at 1745 cm(-)(1) in the C=O stretching region of COOH or ester groups in the Q(B)(-)/Q(B) spectrum. The peak frequency was unaffected by D(2)O substitution, indicating that this peak does not arise from a COOH group but probably from the 10a-ester C=O group of the pheophytin molecule adjacent to Q(B). The absence of protonation of carboxylic amino acids upon Q(B)(-) formation in contrast to the previous observation in the purple bacterium Rhodobacter sphaeroides suggests that the protonation mechanism of Q(B) in PSII is different from that of bacterial reaction centers.     

Formation of 4-hydroxy-2,5-dimethyl-3[2H]-furanone by Zygosaccharomyces rouxii: identification of an intermediate

The formation of the important flavor compound 4-hydroxy-2,5-dimethyl-3[2H]-furanone (HDMF; Furaneol) from D-fructose-1,6-bisphosphate by the yeast Zygosaccharomyces rouxii was studied with regard to the identification of intermediates present in the culture medium. Addition of o-phenylenediamine, a trapping reagent for alpha-dicarbonyls, to the culture medium and subsequent analysis by high-pressure liquid chromatography with diode array detection revealed the formation of three quinoxaline derivatives derived from D-fructose-1,6-bisphosphate under the applied growth conditions (30 degrees C; pH 4 to 5). Isolation and characterization of these compounds by tandem mass spectrometry and nuclear magnetic resonance spectroscopy led to the identification of phosphoric acid mono-(2,3,4-trihydroxy-4-quinoxaline-2-yl-butyl) ester (Q1), phosphoric acid mono-[2,3-dihydroxy-3-(3-methyl-quinoxaline-2-yl)-propyl] ester (Q2), and phosphoric acid mono-[2-hydroxy-3-(3-methyl-quinoxaline-2-yl)-propyl] ester (Q3). Q1 and Q2 were formed independently of Z. rouxii cells, whereas Q3 was detected only in incubation systems containing the yeast. Identification of Q2 demonstrated for the first time the chemical formation of 1-deoxy-2,3-hexodiulose-6-phosphate in the culture medium, a generally expected but never identified intermediate in the formation pathway of HDMF. Since HDMF was detected only in the presence of Z. rouxii cells, additional enzymatic steps were presumed. Incubation of periplasmic and cytosolic protein extracts obtained from yeast cells with D-fructose-1,6-bisphosphate led to the formation of HDMF, implying the presence of the required enzymes in both extracts.     

Arachidonic Acid-metabolizing Cytochrome P450 Enzymes Are Targets of ω-3 Fatty Acids

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) protect against cardiovascular disease by largely unknown mechanisms. We tested the hypothesis that EPA and DHA may compete with arachidonic acid (AA) for the conversion by cytochrome P450 (CYP) enzymes, resulting in the formation of alternative, physiologically active, metabolites. Renal and hepatic microsomes, as well as various CYP isoforms, displayed equal or elevated activities when metabolizing EPA or DHA instead of AA. CYP2C/2J isoforms converting AA to epoxyeicosatrienoic acids (EETs) preferentially epoxidized the ω-3 double bond and thereby produced 17,18-epoxyeicosatetraenoic (17,18-EEQ) and 19,20-epoxydocosapentaenoic acid (19,20-EDP) from EPA and DHA. We found that these ω-3 epoxides are highly active as antiarrhythmic agents, suppressing the Ca²⁺-induced increased rate of spontaneous beating of neonatal rat cardiomyocytes, at low nanomolar concentrations. CYP4A/4F isoforms ω-hydroxylating AA were less regioselective toward EPA and DHA, catalyzing predominantly ω- and ω minus 1 hydroxylation. Rats given dietary EPA/DHA supplementation exhibited substantial replacement of AA by EPA and DHA in membrane phospholipids in plasma, heart, kidney, liver, lung, and pancreas, with less pronounced changes in the brain. The changes in fatty acids were accompanied by concomitant changes in endogenous CYP metabolite profiles (e.g. altering the EET/EEQ/EDP ratio from 87:0:13 to 27:18:55 in the heart). These results demonstrate that CYP enzymes efficiently convert EPA and DHA to novel epoxy and hydroxy metabolites that could mediate some of the beneficial cardiovascular effects of dietary ω-3 fatty acids.