Aspergillus sp136抗细菌耐药活性成分研究

基于广谱抗细菌耐药性这一思路，本研究中心建立了一套抗细菌耐药性化合物的筛选方法。由此从3000多种西南地区特殊生境的微生物和植物样品提取物中筛选获得17个抗细菌耐药性活性样品。对其中一株来自峨嵋山土样的微生物（Aspergillus sp136）进行了深入研究。通过TLC自显影等方法从其发酵产物中追踪分离得到抗耐药有效成分，并鉴定为烟曲霉酸。 采用多种方法对烟曲霉酸的体外抗细菌耐药活性进行评价。在平板扩散法中，烟曲霉酸表现出对青霉素（β-内酰氨抗生素）的协同抗耐药能力，其活性大约3倍于克拉维酸。在MIC的测试实验中，烟曲霉酸表现出对青霉素（β-内酰氨抗生素）以及非β-内酰氨抗生素如红霉素、四环素、氯霉素、链霉素、卡那霉素、庆大霉素的抗耐药能力。在棋盘格杀菌以及时间致死曲线的研究中，烟曲霉酸也表现出对青霉素、红霉素、四环素的协同抗细菌耐药活性。 在广泛的活性筛选中发现烟曲霉酸对LDLR基因具有上调活性，表明烟曲霉酸可能具有降血脂的活性。 在研究中发现，同空白对照相比，烟曲霉酸使耐药菌（Bacillus cereus NCPF63509）细胞外β-内酰胺酶酶活大幅度下降，而细胞内β-内酰胺酶酶活仅略有上升，这表明烟曲霉酸对β-内酰胺酶分泌过程具有抑制作用。 综述了β-内酰胺酶的研究进展。 A two-step agar diffusion method was established to screen wide spectrum synergistic antibacterial agents. By using this method, 17 active samples against antibiotic resistance were discovered from more than 3000 plants and microbes, which were collected from southwest china. One isolate Aspergillus sp136 collected from E-mei mountain area was selected for further studies. From the metabolites of this strain, a synergistic antibacterial compound was isolated by bioautographic TLC assay-guided fractionation and identified as helvolic acid. The synergistic effect of helvolic acid was confirmed by several methods in vitro. The synergistic effect of helvolic acid with penicillin (β-lactam antibiotics) was about 3 times as that of clavulanic acid with penicillin in agar diffusion assay. In MIC studies, helvolic acid exhibited synergistic effects with β-lactam antibiotics such as penicillin and non β-lactam antibiotics such as erythromycin, tetracycline, kanamycin, streptomycin and gentamycin. In checkerboard and time-kill studies, helvolic acid also exhibited synergistic effects with penicillin, erythromycin and tetracycline. In general screen of bioactivities, helvolic acid upregulate LDLR gene, which was indirectly determined by the activity of fluorescent enzyme. Therefore, helvolic acid might have the ability to lower lipid in blood. Compared with blank control, the extracellular β-lactamase activity decrease significantly and the intracellular β-lactamase activity increase slightly in Bacillus cereus NCPF63509 in the presence of helvolic acid, indicating that the secretion of β-lactamase was inhibited by helvolic acid. The research of β-lactamase was reviewed.

Benzodiazepine Alkaloids from Marine-Derived Endophytic Fungus Aspergillus ochraceus

A new fungus-derived benzodiazepine analogue, 2-hydroxycircumdatin C (1), and a compound which has been isolated from a natural resource for the first time, but has been previously synthesized, namely (11aS)-2,3-dihydro-7-methoxy-1H-pyrrolo[2,1-c][1,4]benzodiazepine-5,11(10H,11aH)-dione (2), along with five structurally related known alkaloids (3-7), were isolated from Aspergillus ochraceus, an endophytic fungus derived from the marine brown alga Sargassum kjellmanianum. Their structures were established on the basis of spectroscopic methods. The absolute configuration of I was determined through CD evidence. Compound 1 displayed significant DPPH radical-scavenging activity with an IC50 value of 9.9 mu M, which is 8.9-fold more potent than that of butylated hydroxytoluene (BHT), a well-known synthetic positive control.

A new naphthoquinoneimine derivative from the marine algal-derived endophytic fungus Aspergillus niger EN-13

Cultivation of an endophytic fungus Aspergillus niger EN-13 that was isolated from the inner tissue of the marine brown alga Colpomenia sinuosa resulted in the characterization of a new naphthoquinoneimine derivative, namely, 5,7-dihydroxy-2-[1-(4methoxy-6-oxo-6H-pyran-2-yl)-2-phenylethylaniino]-[1,4]naphthoquinone. The structure of the new compound was established on the basis of various NMR spectroscopic analyses including 2D NMR techniques, EI-MS, and HR-ESI-MS. This compound displayed moderate antifungal activity. (c) 2007 Bin Gui Wang. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Ergosterimide, a new natural Diels-Alder adduct of a steroid and maleimide in the fungus Aspergillus niger

Ergosterimide (1), a natural Diels-Alder adduct of ergosteroid and maleimide, was characterized from the culture extract of Aspergillus niger EN-13, an endophytic fungus isolated from the marine brown alga Colpomenia sinuosa. In addition, four known steroids including (22E,24R)-ergosta-5,7,22-trien-3 beta-ol (2), (22E,24R)-ergosta-4,6,8(14),22-tetraen-3one (3), (22E,24R)-5 alpha,8 alpha-epidioxyergosta-6,22-dien-3 beta-ol (4), and (22E,24R)-ergosta-7,22dien-3 beta,5 alpha,6 beta-triol. (5) were also isolated and identified. The structures of these compounds were elucidated by extensive analysis of 1D and 2D NMR and IR spectra and MS data. The plausible biosynthetic pathway of 1 was also discussed. To the best of our knowledge, 1 is the first natural Diels-Alder adduct of steroid and maleimide reported so far. (C) 2007 Elsevier Inc. All rights reserved.

New sphingolipids with a previously unreported 9-Methyl-C-20-sphingosine moiety from a marine algous endophytic fungus Aspergillus niger EN-13

Asperamides A (1) and B (2), a sphingolipid and their corresponding glycosphingolipid possessing a hitherto unreported 9-methyl-C-20-sphingosine moiety, were characterized from the culture extract of Aspergillus niger EN-13, an endophytic fungus isolated from marine brown alga Colpomenia sinuosa. The structures were elucidated by spectroscopic and chemical methods as (2S,2'R,3R,3'E,4E,8E)-N-(2'-hydroxy-3'-hexadecenoyl)-9-methyl-4,8-icosadien-1,3-diol (1) and 1-O-beta-D-glucopyranosyl-(2S,2'R,3R,3'E,4E,8E)-N-(2'-hydroxy-3'-hexadecenoyl)-9-methyl-4,8-icosadien-1,3-diol (2). In the antifungal assay, asperamide A (1) displayed moderate activity against Candida albicans.

磷矿石的微生物风化作用——以一株黑曲霉（Aspergillus niger）为例

以直接和间接作用的方式研究培养基中黑曲霉Aspergillus niger对磷矿粉的风化作用。在装有100目磷矿石粉的液体培养基中接入该菌研究其对磷矿石粉的直接风化作用;同时将装有100目磷矿石粉的透析袋放入液体培养基中再接入该菌研究其对磷矿石粉的间接风化作用。按不同时间取培养液上清液,用电感耦合等离子体-发射光谱仪测定Ca2+、Mg2+、Al3+、Fe3+、K+和Mn2+浓度,用离子色谱法测定H2PO4-、SO42-和Cl-浓度。此外,黑曲霉风化作用后的矿物残渣用电子探针作表面微观形态分析和XRD矿物物相分析。结果表明:黑曲霉对磷矿石粉风化作用的直接作用强度大于间接作用;

一种海藻内生真菌二萜生物碱类化合物的应用

本发明涉及杀虫剂领域，具体地说是一种海藻内生真菌二萜生物碱类化合物及其制备和应用。具体结构式如（I）所示，其制备方法为将米曲霉（Aspergillus oryzae）cf-2接种于培养基中静止发酵，发酵液经乙酸乙酯萃取浓缩，菌丝体用有机溶剂提取，再经乙酸乙酯萃取浓缩，合并浓缩物，得到粗提物；粗提物进行硅胶柱层析，用有机溶剂进行梯度洗脱，收集洗脱液，洗脱液经薄层层析检测；将以洗脱液体积比5-8：1梯度的洗脱组分进行凝胶柱层析、硅胶柱层析和薄层层析分离纯化得目标化合物。本发明获得的海藻内生真菌二萜生物碱类化合物，经杀虫活性实验得出化合物在100微克/毫升时对卤虫的致死率为42.9%。

一种海藻内生真菌二萜生物碱类化合物及其制备和应用

本发明涉及杀虫剂领域，具体地说是一种海藻内生真菌二萜生物碱类化合物及其制备和应用。具体结构式如（I）所示，其制备方法为将米曲霉（Aspergillus oryzae）cf-2接种于培养基中静止发酵，发酵液经乙酸乙酯萃取浓缩，菌丝体用有机溶剂提取，再经乙酸乙酯萃取浓缩，合并浓缩物，得到粗提物；粗提物进行硅胶柱层析，用有机溶剂进行梯度洗脱，收集洗脱液，洗脱液经薄层层析检测；将以洗脱液体积比10-15：1梯度的洗脱组分进行凝胶柱层析、硅胶柱层析和薄层层析分离纯化得目标化合物。本发明获得的海藻内生真菌二萜生物碱类化合物，经杀虫活性实验得出化合物在100微克/毫升时对卤虫的致死率为61.9%。

海藻内生真菌二萜生物碱类化合物及其制备和应用

本发明涉及杀虫剂领域，具体地说是一种海藻内生真菌二萜生物碱类化合物及其制备和应用。具体结构式如（I）所示，其制备方法为将米曲霉（Aspergillus oryzae）cf-2接种于培养基中静止发酵，发酵液经乙酸乙酯萃取浓缩，菌丝体用有机溶剂提取，再经乙酸乙酯萃取浓缩，合并浓缩物，得到粗提物；粗提物进行硅胶柱层析，用有机溶剂进行梯度洗脱，收集洗脱液，洗脱液经薄层层析检测；将以洗脱液体积比5-8：1梯度的洗脱组分进行凝胶柱层析、硅胶柱层析和薄层层析分离纯化得目标化合物。本发明获得的海藻内生真菌二萜生物碱类化合物，经杀虫活性实验得出化合物在100微克/毫升时对卤虫的致死率为42.9%。

一种天然海藻内生真菌二萜生物碱类化合物及其制备和应用

本发明涉及细菌抑制剂领域，具体地说是一种天然的海藻内生真菌二萜生物碱类化合物及其制备和应用。具体结构式如（I）所示，其制备方法为将米曲霉（Aspergillus oryzae）cf-2接种于培养基中静止发酵，发酵液经乙酸乙酯萃取浓缩，菌丝体先用有机溶剂提取，再经乙酸乙酯萃取浓缩，合并浓缩物，得到粗提物；粗提物进行硅胶柱层析，用有机溶剂进行梯度洗脱，收集洗脱液，洗脱液经薄层层析检测；将以洗脱液体积比0-10：100梯度，洗脱下的组分进行凝胶柱层析、硅胶柱层析和薄层层析分离纯化得目标化合物。本发明所得二萜生物碱类化合物具有显著的抑菌活性。

一种天然海藻内生真菌次生代谢产物二萜生物碱类化合物的应用

本发明涉及杀虫剂领域，具体地说是一种海藻内生真菌次生代谢产物二萜生物碱类化合物的应用。所述海藻内生真菌次生代谢产物二萜生物碱类化合物具有杀虫作用，海藻内生真菌次生代谢产物二萜生物碱类化合物如式（I）所示；本发明通过分离于海洋红藻异管藻的真菌米曲霉（Aspergillus oryzae）cf-2发酵经提取、分离获得的二萜生物碱类天然化合物，经杀虫活性实验得出此二萜生物碱类化合物在100微克/毫升时对卤虫的致死率为74.2%。

稠油废水处理中优势微生物种群组成及变化规律

针对辽河油田锦采污水处理厂稠油废水，利用传统培养方法和PCR-DGGE诊断技术，对稠油废水处理过程中优势微生物种群组成和多样性进行全面系统的研究。结果表明，微生物对稠油废水生物处理的作用为细菌>真菌>放线菌。细菌数量、基因多样性指数与废水中TPH、CODCr均正相关，可以作为稠油废水水质评价的生物指标。 对影响稠油废水生物降解的主要因子进行优化表明，当30℃，pH值7.5，HRT为216h，添加N、P营养盐使N:P比为5.63:1时，CODCr去除率最高，去除后CODCr值满足污水综合排放一级标准（GB8978-1996）。利用GC-MS技术分析降解前后稠油废水中主要有机成分表明，微生物对饱和烃类化合物降解率最高，其次是低分子量芳香烃，而高分子量芳香烃、胶质和沥青质最低。 以稠油为唯一碳源，对筛选出的菌株进行摇瓶实验表明，各菌株对稠油均具有一定的降解能力，其中F0504除油能力最强，56d去除率可达63.3%；动力学方程拟合表明稠油生物降解过程符合一级动力学方程。降解后残油组分分析表明，B0505和F0501对烷烃、B0510、F0505和F0507对芳香烃、B0501和F0504对胶质、沥青质的去除率均较高，去除率都在30c%之间。 经鉴定，优势菌株B0501和B0505分别为液化金杆菌（Aureobaterium liquefaciens）和弗氏丙酸杆菌（Propionibacterium freuclenreichii），主要真菌有青霉（Penicillium）、曲霉（Aspergillus）、木霉（Trichoderma）和交链孢霉（Alternaria）。

Aspergillusterricola角蛋白酶高产菌株选育及酶学性质研究

根据非对称原生质体融合原理，建立了非营养缺陷型原生质体融合模型，丰富了原生质体育种方法.利用此模型进行栖土曲霉（Aspergillusterricola）不同菌株的种内融合，选育出高活性角蛋白酶生产菌.通过生长速度、分生孢子体积、孢子DNA含量的测定，并利用RAPD-PCRDNA指纹分析技术，确定其为杂合二倍体.连续传接12代后，菌株产酶性能稳定.发酵试验表明，其产角蛋白酶活性达到2840Ug<'-1>，分别比双亲的产酶能力提高了42%和30%.构建了栖土曲霉染色体基因组文库，并从文库中筛选出角蛋白酶生物合成基因的阳性克隆，为进一步在分子水平上深入研究角蛋白酶奠定了基础.

土壤-作物体系黄曲霉种群分布及其毒素污染

在辽宁地区开展土壤-作物体系黄曲霉种群分布及其毒素污染研究，结果表明：黄曲霉是辽宁地区土壤和作物当中存在的优势种，L品系是主要品系；在黄曲霉种群构成及黄曲霉品系分布上具有地域特征。土壤黄曲霉菌落数在作物不同生育期具有显著差异。分别从土壤、玉米当中分离到的黄曲霉产毒率是22％和46％，土壤～作物体系中黄曲霉产毒率是39％。试验中未检出强产毒菌株，说明辽宁地区土壤一作物体系中黄曲霉菌株具有较低的产毒能力。依据我国现行的粮食中黄曲霉毒素允许标准，辽宁地区的玉米黄曲霉毒素污染处于低水平，平均黄曲霉毒素含量为0.89μg．kg-1，符合粮食安全标准。但少数样本超出了一些发达国家颁布的严格限定，表明该区仍需对玉米黄曲霉毒素污染加以控制。从区域分布看，干旱较重的辽西地区黄曲霉毒素污染较湿润的辽东和辽中地区严重。黄曲霉毒素污染随着玉米储藏时间延长有增加的趋势。土壤黄曲霉菌落数与玉米黄曲霉毒素污染呈正相关关系，用土壤黄曲霉菌落数能较好地反映该地区的黄曲霉毒素污染状况。轮作对一减少土壤中黄曲霉数量具有显著影响。在田间管理中应当实施合理轮作及适时灌溉，可预防黄曲霉侵染及降低黄曲霉毒素污染。