来自普哥滨珊瑚的一株Talaromyces sp.真菌的活性次生代谢产物

利用硅胶柱层析、制备型HPLC和重结晶等手段从普哥滨珊瑚分离的一株Talaromyces sp.真菌C21-1中筛选得到2个活性化合物,运用核磁共振、质谱和圆二色谱等技术鉴定这两个化合物分别为(R)-(-)-hydroxysydonic acid（1）和homodimeric WIN 64821（2）,补充完善了化合物2的核磁共振信号归属,并对化合物进行抗菌活性和乙酰胆碱酯酶抑制活性的测定,发现化合物1对白色假丝酵母Canidia albicans和耐甲氧基青霉素的金黄色葡萄球菌Methicillin-resistant Staphylococcus aureus（MRSA）具有一定的抑制作用,其最低抑菌浓度（MIC）分别为0.075mmol/L和0.2mmol/L,对副溶血弧菌Vibrio parahemolyticus的抑制活性较弱,在0.2mmol/L浓度下的抑制率为17%;化合物2最大浓度0.2mmol/L条件下对这3种菌均没有明显的抑制效果。化合物2表现出剂量依赖的乙酰胆碱酯酶抑制活性,0.5mmol/L时抑制率达到35.1%。     

水葫芦内生拟枝孢镰刀菌SC1608的次生代谢产物

研究了一株水生植物水葫芦内生拟枝孢镰刀菌Fusarium sporotrichioides SC1608大米发酵物的抗真菌活性次生代谢产物,从中共分离获得6个化合物,经高分辨质谱、三重四极杆质谱和核磁共振谱分析,分别鉴定为4个鞘糖脂类化合物：asperamide B (1)、asperamide D (2)、asperamide E (3)、cerebrosides A (4),以及2个环酯肽类化合物acuminatum A (5)和acuminatum B (6),其中化合物2和化合物3是新的鞘糖脂类化合物。通过滤纸片琼脂扩散法评价了单体化合物的抗真菌活性,证实化合物5和6对柑橘绿霉和玉米小斑病菌均具有较强的体外抑制活性。研究结果初步揭示了F. sporotrichioides SC1608发酵物的抗真菌活性物质基础,丰富了拟枝孢镰刀菌次生代谢产物的化学多样性。     

海洋真菌抗病原菌生物活性物质的研究

对分离自大连海域的海洋生物共附生真菌进行了抗菌活性的初筛与复筛,发现有6株具有抗表皮葡萄球菌和铜绿假单胞菌的活性,发现培养基成分及培养方式对活性菌株抗菌物质的产生有显著影响。菌株11-N2抗菌活性最强,其发酵液抗表皮葡萄球菌的效价相当于为氨苄青霉素11.4 U/mL,抗铜绿假单胞菌的效价相当于氨苄青霉素6.0 U/mL。通过硅胶柱层析对该菌株提取物的抗菌活性成分进行了初步追踪,发现二氯甲烷∶甲醇=15∶1洗脱组分具有最强活性,薄层层析显示该组分主要成分在254 nm紫外光下具有强吸收,薄层显色反应提示其主要成分可能为含胺基的甾体或三萜类、有机胺类、吲哚衍生物类化合物。     

不同培养期深色有隔内生真菌链格孢菌对紫花苜蓿生长的影响

接种深色有隔内生真菌（dark septate endophytes,DSE）可以提高植物对养分的吸收利用,促进植物生长发育。为探讨接种不同培养时间的DSE链格孢菌Alternaria sp. CGMCC 17463对紫花苜蓿生长的影响,以紫花苜蓿为研究对象,接种培养4、6、8、10d的Alternaria sp. CGMCC 17463鲜样菌剂和风干样菌剂进行盆栽试验,以阐明不同培养时间的Alternaria sp. CGMCC 17463对紫花苜蓿出芽率、生长特性和营养含量的作用。结果表明,接种Alternaria sp. CGMCC 17463鲜样菌剂和风干样菌剂的紫花苜蓿根系内均有Alternaria sp. CGMCC 17463定殖,且均能增加紫花苜蓿株高、地上部干重、根部干重和根冠比。其中,接种培养10d的Alternaria sp. CGMCC 17463鲜样菌剂和培养8d的风干样菌剂对紫花苜蓿的促生效果最佳。同时,接种鲜样菌剂和风干样菌剂可以提高紫花苜蓿对养分吸收。随Alternaria sp. CGMCC 17463培养时间的增加,接种鲜样菌剂和风干样菌剂的紫花苜蓿的氮、磷、钾含量均呈先增加再降低的趋势。接种培养4-6d的Alternaria sp. CGMCC 17463菌剂,紫花苜蓿的氮、磷、钾含量相对较高。Pearson 相关性分析表明,Alternaria sp. CGMCC 17463培养时间与紫花苜蓿株高、根干重、根系Alternaria sp. CGMCC 17463总侵染率和菌丝侵染率呈显著正相关（P<0.05）,菌剂类型不是影响紫花苜蓿生长的主要因素,本研究为进一步开发利用Alternaria sp. CGMCC 17463菌剂提供了理论支撑。     

滇南红厚壳种子油的脂肪酸成分

应用GC-MS联用技术对云南省西双版纳产的滇南红厚壳（Calophyllumpolyanthum）种子油的脂肪酸成分进行分析.检出12种脂肪酸成分,占总量的99.39%,主要是亚油酸（38.75%）,棕榈酸（22.42%）,油酸（22.11%）和硬脂酸（9.81%）.     

拮抗临床致病菌的一株地衣内生真菌

本研究运用组织分离法对采自贵州省都匀市斗篷山景区的地衣进行内生真菌分离,乙酸乙酯萃取菌株液体发酵物,通过打孔法筛选地衣内生真菌提取物并对9株临床致病菌株及耐药菌株进行抑菌活性实验。结果表明一株分离自星点梅衣属地衣Punctelia sp.的内生真菌DPS-165-9对其中的6株显示抑制活性。此6株菌为金黄色葡萄球菌Staphylococcus aureus 25923和6538、耐药表面葡萄球菌Staphylococcus epidermidis Z12、耐药布氏杆菌Brucella sp. B103、耐药枯草芽孢杆菌Bacillus subtilis 163和耐药藤黄微球菌Micrococcus luteus 261。经鉴定DPS-165-9为蔡氏轮层炭壳菌Daldinia childiae。本研究为下一步挖掘新的抗菌物质奠定基础。     

青霉属真菌Penicillium sp. CPCC 400786的抗病毒活性成分

采用抗艾滋病毒抑制剂筛选模型对一株青霉属真菌Penicillium sp. CPCC 400786发酵产物的乙酸乙酯提取物进行活性评价,结果显示,其对艾滋病毒有较强的抑制活性。采用正相硅胶柱、Sephadex LH-20凝胶柱和半制备HPLC等色谱技术对乙酸乙酯提取物进行分离纯化,从中分离得到8个化合物。通过波谱数据分析,分别鉴定为：oxalicine A（1）、oxalicine B（2）、cis-4,6-dihydroxymellein（3）、亚油酸（4）、十八烯酸（5）、肉豆蔻酸（6）、尿嘧啶（7）、胸腺嘧啶（8）。化合物1和2为杂萜类化合物。对化合物1-6进行了抗艾滋病毒（HIV-1）和抗甲型流感病毒（H1N1）的活性评价。结果显示,化合物1具有良好的抗H1N1活性,其IC₅₀值为38.5μmol/L,比阳性对照药利巴韦林稍弱（IC₅₀=20.5μmol/L）;化合物1和2具有抗HIV-1的活性,其IC₅₀值分别为22.4、67.8μmol/L;其他化合物未显示抗病毒活性。本研究为从青霉属中发现更多抗病毒活性杂萜分子提供了依据。     

原始热带雨林鹦歌岭土壤抗MRSA放线菌的分离与筛选

鹦歌岭是位于海南省乐东县的原始热带雨林,从鹦歌岭土壤中分离与筛选抗耐甲氧西林金黄色葡萄球菌(MRSA)的放线菌。采用平板稀释法分离,以MRSA作为受试菌株,测试抗菌活性,对具有抗菌活性的菌株进行16S rDNA序列测定。采用滤纸片法,测定菌株发酵粗提液对MRSA抑菌活性,并用高效液相色谱仪检测粗提液的活性物质。从鹦歌岭土壤中共分离到168株放线菌,其中10株具有较强的抗MRSA活性,发酵液粗提物抑菌圈直径在8 mm-25 mm之间;16S rDNA序列比对发现,10株菌均与链霉菌属(Streptomyces)的相似性达99%以上,初步确定是链霉菌属的放线菌。通过对比10株菌发酵液HPLC指纹图谱,发现菌株7、5和9-1的发酵液次级代谢产物丰富,活性测试表明,10株放线菌对MRSA的生长均有明显的抑制作用。     

共轭亚油酸甘油酯对大鼠血清脂肪酸组成影响的研究

目的：研究共轭亚油酸甘油酯对高脂饮食大鼠血清脂肪酸组成变化的影响;方法：60 只SD 大鼠随机分为5 组：正常对照 组、高脂对照组,共轭亚油酸甘油酯低（2 g/kg·bw）、中（4 g/kg·bw）、高（6 g/kg·bw）剂量组,除基础对照组外其余各组均喂饲高脂 饲料,建立高脂模型,以灌胃方式给予受试物,6 周后取血清测定其脂肪酸组成。采用一步法直接对血清中脂肪酸进行甲酯化,气 相色谱毛细管柱分离,氢火焰离子化检测器（FID）定性定量分析;结果：共轭亚油酸甘油酯低、中、高剂量组大鼠血清中单不饱和 脂肪酸含量为25.66%,18.74%,17.72%,与高脂对照组相比显著性下降（P<0.01）,各剂量组饱和脂肪酸和多不饱和脂肪酸含量分 别为48.08%,48.52%,51.15%和27.03%,29.28%,31.13%,与高脂对照组相比显著性升高（P<0.01）。几种代表性脂肪酸如各剂量 组中的油酸、亚油酸与高脂对照组相比分别增加了26.48%,41.56%,51.26% 和9.18%,8.61%,8.73%,各剂量组中棕榈酸与高脂对 照组相比降低了5.28%, 8.80%, 10.92%。结论：共轭亚油酸甘油酯能够增加大鼠血清中饱和脂肪酸和多不饱和脂肪酸的含量,减 少单不饱和脂肪酸含量,改变血清脂肪酸组成。     

细果角茴香内生真菌X-396的分离鉴定及其发酵物脂溶性成分分析

以藏药细果角茴香叶为材料,采用组织分离法分离纯化出一株内生真菌X-396。经形态学观察、ITS测序与序列比对鉴定内生真菌X-396为间座壳属(Diaporthe sp.)真菌Diaporthe sp.YG-2015。进一步利用气相色谱-质谱(GC-MS)联用技术对该真菌发酵物脂溶性成分进行检测与分析,并通过CCK-8法测定其抗肿瘤活性。结果表明,发酵物脂溶性成分共鉴定出35个,其中含量较高的成分为麦角甾醇(31.83%)、β-谷甾醇(12.00%)和十六酸甲酯(4.22%);体外抗肿瘤活性测试显示,发酵物脂溶性成分对人肿瘤细胞株A549、MCF-7和HepG2均具有不同程度的抑制作用,其中对HepG2抑制效果最好,IC₅₀为22.65μg/mL。本研究表明细果角茴香内生真菌X-396发酵物脂溶性成分中具有产生抗肿瘤活性化合物的潜力,具有进一步深入研究的价值。     

Production and biological activity of laidlomycin, anti-MRSA/VRE antibiotic from Streptomyces sp. CS684

Culture broth of a streptomycete isolate, Streptomyces sp. CS684 showed antibacterial activity on methicillin resistant Staphylococcus aureus (MRSA) and vancomycin resistant enterococci (VRE). Among purified substances from the organism, CSU-1, which is active against MRSA and VRE, is a C37H62O12Na (M+, 721.3875), and identified as laidlomycin. The anti-MRSA and anti-VRE activity of CSU-1 was stronger than oxacillin and vancomycin. Phylogenetic analysis showed that strain CS684 is very similar to Streptomyces ardus NRRL 2817T, whereas the ability of Streptomyces sp. CS684 to produce laidlomycin was shown to be unique.     

Antibiotic effect of linolenic acid fromChlorococcum strain HS-101 andDunaliella primolecta on methicillin-resistantStaphylococcus aureus

Methanol extracts fromChlorococcum strain HS-101 andDunaliella primolecta strongly inhibited the growth of a strain of methicillin-resistantStaphylococcus aureus (MRSA), which is causing serious problems in Japanese hospitals. So that the anti-MRSA substance(s) could be purified and identified, the growth medium was improved for antibiotic production. When the two strains were cultured in their improved media, antibiotic production byChlorococcum strain HS-101 was 1.8-fold that in the standard BG-11 medium, and production byD. primolecta was 2.3-fold. The activity pattern of fractions eluted by silica-gel or gel-permeation chromatography suggested that both strains produced two antibiotic substances. Identification of the purified substances by NMR and GC-MS showed that one of the active substances in both strains was-linolenic acid. Ten fatty acids from other sources were tested, and it was found that unsaturated fatty acids had antibiotic activity against MRSA, with the highest activity that of -linolenic acid.     

The impact of fatty acids on the antibacterial properties of N-thiolated β-lactams

Bacterial fatty acid synthesis (FAS) is a potentially important, albeit controversial, target for antimicrobial therapy. Recent studies have suggested that the addition of exogenous fatty acids (FAs) to growth media can circumvent the effects of FAS-targeting compounds on bacterial growth. Consequently, such agents may have limited in vivo applicability for the treatment of human disease, as free FAs are abundant within the body. Our group has previously developed N-thiolated β-lactams and found they function by interfering with FAS in select pathogenic bacteria, including MRSA. To determine if the FAS targeting activity of N-thiolated β-lactams can be abrogated by exogenous fatty acids, we performed MIC determinations for MRSA strains cultured with the fatty acids oleic acid and Tween 80. We find that, whilst the activity of the known FAS inhibitor triclosan is severely compromised by the addition of both oleic acid and Tween 80, exogenous FAs do not mitigate the antibacterial activity of N-thiolated β-lactams towards MRSA. Consequently, we propose that N-thiolated β-lactams are unique amongst FAS-inhibiting antimicrobials, as their effects are unimpeded by exogenous FAs.     

Thematic review series: skin lipids. Antimicrobial lipids at the skin surface

The skin surface represents our interface with the external environment, and as such, is our first line of defense against microbial colonization and infection. Lipids at the skin surface are thought to underlie at least part of an antimicrobial barrier. Some of these lipids are synthesized in the epidermis and are carried to the surface as cells differentiate, whereas others are secreted onto the surface from the sebaceous glands. One such group, free sphingoid bases, are known to have broad antimicrobial activity, and our previous studies demonstrate their presence at the skin surface. Free sphingoid bases may be generated by enzymatic hydrolysis of epidermal ceramides. In addition, our preliminary results demonstrate potent antibacterial activity associated with two specific fatty acids derived from sebaceous triglycerides. Most remarkably, one of these fatty acids (sapienic acid, C16:1Delta6), in combination with a low concentration of ethanol, is very effective against methicillin-resistant Staphylococcus aureus (MRSA). In fact, this combination was far more effective than mupirocin with or without ethanol. Mupirocin is a "gold standard" for activity against MRSA.     

Antibacterial properties of a new isoflavonoid from Erythrina poeppigiana against methicillin-resistant Staphylococcus aureus.

A new isoflavonoid, together with four known isoflavonoids, was isolated from the roots of Erythrina poeppigiana. The chemical structure was determined by extensive spectroscopic studies, and then its antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) was investigated. The new isoflavonoid was identified as 3,9-dihyroxy-10-gamma,gamma-dimethylallyl-6a,11a-dehydropterocarpan (compound 1). Compound 1 inhibited bacterial growth most potently of the five isolates, and had a minimum inhibitory concentration (MIC) of 125 microg/ml against thirteen MRSA strains. Inhibitory activity was based on bactericidal action and viable cell number reduced by approximately 1/10,000 after 4 h incubation with compound 1. Despite intense bactericidal action against MRSA, compound 1 never resulted in leakage of 260 nm-absorbing substances from bacterial cells. Compound 1 (12.5 microg/ml) completely inhibited incorporation of radio-labeled thymidine, uridine and leucine into MRSA cells. Although glucose incorporation was also markedly inhibited by the compound, the amount of glucose incorporated by bacterial cells increased gradually with incubation time. These findings suggest that compound 1 exhibits anti-MRSA activity by interfering with incorporation of metabolites and nutrients into bacterial cells or by affecting the nucleic acids of MRSA cells. Furthermore, this new compound could be a potent phytotherapeutic agent for treating MRSA infections.     

Structure-activity relationships of pyrazole-4-carbodithioates as antibacterials against methicillin–resistant Staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of serious hospital-acquired infections and is responsible for significant morbidity and mortality in residential care facilities. New agents against MRSA are needed to combat rising resistance to current antibiotics. We recently reported 5-hydroxy-3-methyl-1-phenyl-1H-pyrazole-4-carbodithioate (HMPC) as a new bacteriostatic agent against MRSA that appears to act via a novel mechanism. Here, twenty nine analogs of HMPC were synthesized, their anti-MRSA structure-activity relationships evaluated and selectivity versus human HKC-8 cells determined. Minimum inhibitory concentrations (MIC) ranged from 0.5 to 64?μg/mL and up to 16-fold selectivity was achieved. The 4-carbodithioate function was found to be essential for activity but non-specific reactivity was ruled out as a contributor to antibacterial action. The study supports further work aimed at elucidating the molecular targets of this interesting new class of anti-MRSA agents.     

CP6679, a new injectable cephalosporin. Part 1: synthesis and structure-activity relationships

A series of cephalosporins bearing a 5,5-fused ring system, an (imidazo[5,1-b]thiazolium-6-yl)methyl group, at the C-3 position were synthesized and evaluated for in vitro antibacterial activities. CP6679 (1s) and its analogues showed potent antibacterial activities against Gram-positive and Gram-negative bacteria, including Pseudomonas aeruginosa. They were also highly active against methicillin-resistant Staphylococcus aureus (MRSA). CP6679 (1s) showed more potent antibacterial activity than ceftazidime (CAZ) or cefpirome (CPR) against Pseudomonas aeruginosa and MRSA.     

Fatty acid synthesis is a target for antibacterial activity of unsaturated fatty acids

Long-chain unsaturated fatty acids, such as linoleic acid, show antibacterial activity and are the key ingredients of antimicrobial food additives and some antibacterial herbs. However, the precise mechanism for this antimicrobial activity remains unclear. We found that linoleic acid inhibited bacterial enoyl-acyl carrier protein reductase (FabI), an essential component of bacterial fatty acid synthesis, which has served as a promising target for antibacterial drugs. Additional unsaturated fatty acids including palmitoleic acid, oleic acid, linolenic acid, and arachidonic acid also exhibited the inhibition of FabI. However, neither the saturated form (stearic acid) nor the methyl ester of linoleic acid inhibited FabI. These FabI-inhibitory activities of various fatty acids and their derivatives very well correlated with the inhibition of fatty acid biosynthesis using [(14)C] acetate incorporation assay, and importantly, also correlated with antibacterial activity. Furthermore, the supplementation with exogenous fatty acids reversed the antibacterial effect of linoleic acid, which showing that it target fatty acid synthesis. Our data demonstrate for the first time that the antibacterial action of unsaturated fatty acids is mediated by the inhibition of fatty acid synthesis.     

A Fatty Acid from the Diatom Phaeodactylum tricornutum is Antibacterial Against Diverse Bacteria Including Multi-resistant Staphylococcus aureus (MRSA)

Pathogenic bacteria, such as multidrug-resistant Staphylococcus aureus (MRSA), which are not susceptible to most conventional antibiotics, are causing increased concern in healthcare institutions worldwide. The discovery of novel antibacterial compounds for biomedical exploitation is one avenue that is being pursued to combat these problematic bacteria. Marine eukaryotic microalgae are known to produce numerous useful products but have attracted little attention in the search for novel antibiotic compounds. Cell lysates of the marine diatom, Phaeodactylum tricornutum Bohlin, have been reported to display antibacterial activity in vitro, but the compounds responsible have not been fully identified. In this paper, using column chromatography and reversed-phase high-performance liquid chromatography, we report the isolation of an antibacterial fatty acid. Mass spectrometry and ¹H-nuclear magnetic resonance spectroscopy revealed it to be the polyunsaturated fatty acid, eicosapentaenoic acid (EPA). We show that EPA is active against a range of both Gram-positive and Gram-negative bacteria, including MRSA, at micromolar concentrations. These data indicate that it could find application in the topical and systemic treatment of drug-resistant bacterial infections.     

Linoleic acid, α-linolenic acid,and monolinolenins as antibacterial substances in the heat-processed soybean fermented with Rhizopus oligosporus

ABSTRACT

Antibacterial activities against Staphylococcus aureus and Bacillus subtilis were found in an ethanol fraction of tempe, an Indonesian fermented soybean produced using Rhizopus oligosporus. The ethanol fraction contained free fatty acids, monoglycerides, and fatty acid ethyl esters. Among these substances, linoleic acid and α-linolenic acid exhibited antibacterial activities against S. aureus and B. subtilis, whereas 1-monolinolenin and 2-monolinolenin exhibited antibacterial activity against B. subtilis. The other free fatty acids, 1-monoolein, monolinoleins, ethyl linoleate, and ethyl linolenate did not exhibit bactericidal activities. These results revealed that R. oligosporus produced the long-chain polyunsaturated fatty acids and monolinolenins as antibacterial substances against the Gram-positive bacteria during the fungal growth and fermentation of heat-processed soybean.